Molecular and Cellular Probes最新文献_第7页

Botox-A induced apoptosis and suppressed cell proliferation in fibroblasts pre-treated with breast cancer exosomes 肉毒杆菌a诱导乳腺癌外泌体预处理成纤维细胞凋亡和抑制细胞增殖。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2025-01-06 DOI: 10.1016/j.mcp.2024.102007

Hossein Sayaf , Niloufar Salimian , Mahnaz Mohammadi , Parisa Ahmadi , Amir Gholamzad , Sadegh Babashah , Maliheh Entezari , Najma Farahani , Maryam Montazeri , Mehrdad Hashemi

Background

breast cancer-associated fibroblast (CAF) is linked to metastasis and is poor for breast cancer prognosis. Since Clostridium Toxin A (Botox-A) had represented a cytotoxic effect on fibroblasts, this study aims to assess Botox-A cytotoxicity in both normal fibroblasts and exosome-induced CAFs.

Material and method

the serum exosomes of 40 BC patients and 30 healthy individuals were isolated and lncRNA H19 (lnch19) levels were assessed by qRT-PCR method. After that, Breast Cancer (BC) exosomes co-cultured with Human foreskin fibroblasts (HFF) and qRT-PCR were applied to evaluate α-SMA, Vimentin, BCL-2, and BAX expression. Both Normal and malignant HFFs co-cultured with Botox-A, and Botox-A loaded exosome for 24 and 48 h and their apoptosis, Cell proliferation, and viability were monitored by MTT assay, Annexin V-FITC and PI staining and qRT-PCR for BCL-2, BAX, and cyclin D1 mRNAs.

Results

Serum exosomes of BC patients had significantly higher levels of lncRNA H19 than healthy individuals. MTT assay results showed Botox-A decreased vital Human foreskin fibroblasts in a dose-dependent manner. BC exosomes significantly increased α-SMA, Vimentin, and BCL-2 mRNA levels in Human foreskin fibroblasts, on the other hand, BAX decreased meaningfully. Co-culture of exosome-treated HFF cells with both Botox-A and Botox-A loaded exosomes significantly boosted BCL-2 mRNA levels, completely contrary to BAX and cyclid d1 expression. Meanwhile, flow cytometry results confirmed a high rate of apoptosis in malignant Human foreskin fibroblasts treated with Botox-A loaded exosome.

Conclusion

The findings of this study indicate that exosomal lncRNA H19 could be a diagnostic marker for Breast Cancer and these Breast cancer exosomes can induce malignant phenotype in fibroblasts and turn them into CAFs. Botox-A could be toxic for both normal fibroblasts and CAFs, inducing apoptosis and suppressing cell proliferation among them.

背景：乳腺癌相关成纤维细胞（CAF）与乳腺癌转移有关，对乳腺癌预后不良。由于肉毒杆菌毒素A （Botox-A）对成纤维细胞具有细胞毒性作用，本研究旨在评估肉毒杆菌A对正常成纤维细胞和外泌体诱导的CAFs的细胞毒性。材料与方法：分离40例BC患者和30例健康人的血清外泌体，采用qRT-PCR法检测lncRNA H19 （lnch19）水平。之后，采用人包皮成纤维细胞（HFF）与乳腺癌（BC）外泌体共培养，采用qRT-PCR检测α-SMA、Vimentin、BCL-2和BAX的表达。用MTT法、Annexin V-FITC和PI染色以及BCL-2、BAX和cyclin D1 mrna的qRT-PCR检测正常和恶性HFFs与Botox-A和负载Botox-A的外泌体共培养24和48小时，并监测其凋亡、细胞增殖和活力。结果：BC患者血清外泌体lncRNA H19水平明显高于健康人。MTT试验结果显示肉毒杆菌a以剂量依赖的方式降低人包皮成纤维细胞。BC外泌体显著提高人包皮成纤维细胞α-SMA、Vimentin和BCL-2 mRNA水平，BAX显著降低。外泌体处理的HFF细胞与装载Botox-A和Botox-A的外泌体共培养显著提高了BCL-2 mRNA水平，与BAX和cycle1的表达完全相反。同时，流式细胞术结果证实，负载Botox-A的外泌体对恶性包皮成纤维细胞有较高的凋亡率。结论：本研究结果提示外泌体lncRNA H19可作为乳腺癌的诊断标志物，这些乳腺癌外泌体可诱导成纤维细胞的恶性表型并将其转化为cas。肉毒杆菌a对正常成纤维细胞和CAFs均有毒性，诱导细胞凋亡，抑制细胞增殖。

{"title":"Botox-A induced apoptosis and suppressed cell proliferation in fibroblasts pre-treated with breast cancer exosomes","authors":"Hossein Sayaf , Niloufar Salimian , Mahnaz Mohammadi , Parisa Ahmadi , Amir Gholamzad , Sadegh Babashah , Maliheh Entezari , Najma Farahani , Maryam Montazeri , Mehrdad Hashemi","doi":"10.1016/j.mcp.2024.102007","DOIUrl":"10.1016/j.mcp.2024.102007","url":null,"abstract":"<div><h3>Background</h3><div>breast cancer-associated fibroblast (CAF) is linked to metastasis and is poor for breast cancer prognosis. Since Clostridium Toxin A (Botox-A) had represented a cytotoxic effect on fibroblasts, this study aims to assess Botox-A cytotoxicity in both normal fibroblasts and exosome-induced CAFs.</div></div><div><h3>Material and method</h3><div>the serum exosomes of 40 BC patients and 30 healthy individuals were isolated and lncRNA H19 (lnch19) levels were assessed by qRT-PCR method. After that, Breast Cancer (BC) exosomes co-cultured with Human foreskin fibroblasts (HFF) and qRT-PCR were applied to evaluate α-SMA, Vimentin, BCL-2, and BAX expression. Both Normal and malignant HFFs co-cultured with Botox-A, and Botox-A loaded exosome for 24 and 48 h and their apoptosis, Cell proliferation, and viability were monitored by MTT assay, Annexin V-FITC and PI staining and qRT-PCR for BCL-2, BAX, and cyclin D1 mRNAs.</div></div><div><h3>Results</h3><div>Serum exosomes of BC patients had significantly higher levels of lncRNA H19 than healthy individuals. MTT assay results showed Botox-A decreased vital Human foreskin fibroblasts in a dose-dependent manner. BC exosomes significantly increased α-SMA, Vimentin, and BCL-2 mRNA levels in Human foreskin fibroblasts, on the other hand, BAX decreased meaningfully. Co-culture of exosome-treated HFF cells with both Botox-A and Botox-A loaded exosomes significantly boosted BCL-2 mRNA levels, completely contrary to BAX and cyclid d1 expression. Meanwhile, flow cytometry results confirmed a high rate of apoptosis in malignant Human foreskin fibroblasts treated with Botox-A loaded exosome.</div></div><div><h3>Conclusion</h3><div>The findings of this study indicate that exosomal lncRNA H19 could be a diagnostic marker for Breast Cancer and these Breast cancer exosomes can induce malignant phenotype in fibroblasts and turn them into CAFs. Botox-A could be toxic for both normal fibroblasts and CAFs, inducing apoptosis and suppressing cell proliferation among them.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 102007"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142899949","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High expression of ARPC1B promotes the proliferation and apoptosis of clear cell renal cell carcinoma cells, leading to a poor prognosis ARPC1B的高表达促进透明细胞肾细胞癌细胞的增殖和凋亡，导致预后不良。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2025-01-18 DOI: 10.1016/j.mcp.2025.102011

Hongbo Wang , Zhendong Liu , Yuelin Du , Xingbo Cheng , Shanjun Gao , Wenjia Liang , Qingyun Zhu , Zhengfa Jiang , Yanzheng Gao , Panfeng Shang

Background

ARPC1B has been identified as a key regulator of malignant biological behavior in various tumors. However, its specific role in clear cell renal cell carcinoma (ccRCC) remains poorly understood. This study aims to evaluate the influence of ARPC1B on the prognosis and disease progression in ccRCC patients.

Methods

Multi-omics data and clinical information from public databases were analyzed to determine the associations between ARPC1B and prognosis, clinical features, immune microenvironment, and drug sensitivity in ccRCC. Co-expression and gene set enrichment analyses were conducted to elucidate the potential role of ARPC1B in ccRCC pathogenesis. Functional assays, including RT-qPCR, CCK8 assays, colony formation assays, immunofluorescence, immunohistochemistry, and xenograft tumor formation in nude mice, were performed to assess ARPC1B's impact on cell proliferation and apoptosis. Flow cytometry and Western blotting were further employed to investigate the underlying molecular mechanisms of ARPC1B in ccRCC.

Results

ARPC1B expression was significantly elevated in ccRCC and associated with an unfavorable prognosis. Both independent and meta-analyses confirmed that ARPC1B is an independent prognostic risk factor in ccRCC. Furthermore, ARPC1B expression significantly correlated with the immune microenvironment and drug sensitivity. In vitro, experiments demonstrated that ARPC1B knockdown suppressed ccRCC cell proliferation and induced apoptosis through the BAX-Bcl-2/c-caspase3/c-PARP axis, which was further validated by in vivo studies.

Conclusion

ARPC1B overexpression is associated with poor prognosis, altered immune status, and drug sensitivity in ccRCC. Furthermore, ARPC1B promotes the malignant behavior of ccRCC cells and holds potential as a prognostic biomarker and therapeutic target for ccRCC.

背景：ARPC1B已被确定为多种肿瘤恶性生物学行为的关键调节因子。然而，其在透明细胞肾细胞癌（ccRCC）中的具体作用仍知之甚少。本研究旨在评估ARPC1B对ccRCC患者预后和疾病进展的影响。方法：分析多组学数据和公共数据库的临床信息，确定ARPC1B与ccRCC患者预后、临床特征、免疫微环境和药物敏感性的关系。通过共表达和基因集富集分析来阐明ARPC1B在ccRCC发病机制中的潜在作用。通过RT-qPCR、CCK8测定、集落形成测定、免疫荧光、免疫组织化学和裸鼠异种移植肿瘤形成等功能分析来评估ARPC1B对细胞增殖和凋亡的影响。通过流式细胞术和Western blotting进一步研究ARPC1B在ccRCC中的潜在分子机制。结果：ARPC1B在ccRCC中表达显著升高，且与不良预后相关。独立分析和荟萃分析均证实ARPC1B是ccRCC的独立预后危险因素。此外，ARPC1B表达与免疫微环境和药物敏感性显著相关。体外实验表明，ARPC1B敲低可通过BAX-Bcl-2/c-caspase3/c-PARP轴抑制ccRCC细胞增殖并诱导凋亡，体内实验进一步验证了这一结论。结论：ARPC1B过表达与ccRCC患者预后不良、免疫状态改变及药物敏感性相关。此外，ARPC1B促进ccRCC细胞的恶性行为，并具有作为ccRCC预后生物标志物和治疗靶点的潜力。

{"title":"High expression of ARPC1B promotes the proliferation and apoptosis of clear cell renal cell carcinoma cells, leading to a poor prognosis","authors":"Hongbo Wang , Zhendong Liu , Yuelin Du , Xingbo Cheng , Shanjun Gao , Wenjia Liang , Qingyun Zhu , Zhengfa Jiang , Yanzheng Gao , Panfeng Shang","doi":"10.1016/j.mcp.2025.102011","DOIUrl":"10.1016/j.mcp.2025.102011","url":null,"abstract":"<div><h3>Background</h3><div>ARPC1B has been identified as a key regulator of malignant biological behavior in various tumors. However, its specific role in clear cell renal cell carcinoma (ccRCC) remains poorly understood. This study aims to evaluate the influence of ARPC1B on the prognosis and disease progression in ccRCC patients.</div></div><div><h3>Methods</h3><div>Multi-omics data and clinical information from public databases were analyzed to determine the associations between ARPC1B and prognosis, clinical features, immune microenvironment, and drug sensitivity in ccRCC. Co-expression and gene set enrichment analyses were conducted to elucidate the potential role of ARPC1B in ccRCC pathogenesis. Functional assays, including RT-qPCR, CCK8 assays, colony formation assays, immunofluorescence, immunohistochemistry, and xenograft tumor formation in nude mice, were performed to assess ARPC1B's impact on cell proliferation and apoptosis. Flow cytometry and Western blotting were further employed to investigate the underlying molecular mechanisms of ARPC1B in ccRCC.</div></div><div><h3>Results</h3><div>ARPC1B expression was significantly elevated in ccRCC and associated with an unfavorable prognosis. Both independent and meta-analyses confirmed that ARPC1B is an independent prognostic risk factor in ccRCC. Furthermore, ARPC1B expression significantly correlated with the immune microenvironment and drug sensitivity. In vitro, experiments demonstrated that ARPC1B knockdown suppressed ccRCC cell proliferation and induced apoptosis through the BAX-Bcl-2/c-caspase3/c-PARP axis, which was further validated by in vivo studies.</div></div><div><h3>Conclusion</h3><div>ARPC1B overexpression is associated with poor prognosis, altered immune status, and drug sensitivity in ccRCC. Furthermore, ARPC1B promotes the malignant behavior of ccRCC cells and holds potential as a prognostic biomarker and therapeutic target for ccRCC.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 102011"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143014908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Targeting the HLC-1, LC-2/ad, and PC-14 lung cancer cell lines by the silver nanoparticles green-formulated by Descurainia sophia leaf extract 以Descurainia sophia叶提取物绿色配制的银纳米粒子靶向HLC-1、LC-2/ad和PC-14肺癌细胞株。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2024-12-17 DOI: 10.1016/j.mcp.2024.102001

Jianjun Ge, Jianbo Wen, Mingjun Jiang, Kefeng Huang, Saichun Qi, Wei Huang, Linlin Tan

Descurainia sophia, as an an ethno-medicinal plant, contains antioxidant compounds that safeguard cellular integrity against various forms of damage and may play a role in cancer prevention. Antioxidant compounds present in this plant facilitate the body's production of new cells and diminish the risk of colon cancer. In recent years, silver nanoparticles synthesized through green methods using ethnomedicinal herbs have been employed in cancers treatment. We have conducted an investigation into silver nanoparticles that were synthesized through green chemistry principles, utilizing the Descurainia sophia leaves extract for lung carcinoma treatment. The efficacy of Ag NPs against prevalent lung cancer cells was assessed. The green-synthesized silver nanoparticles characterization was conducted utilizing X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), ultraviolet–visible spectroscopy (UV–Vis), energy-dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM). The findings from morphological analyses validate the nanoparticles spherical shape, which ranges in size from 20 to 60 nm. The IC₅₀ values were determined to be 173, 125, and 109 μg/mL for HLC-1, LC-2/ad, and PC-14 cell lines, respectively. According to recent data, Ag NPs may be a useful option to support the treatment of lung cancer. Although the current study presents encouraging findings, further investigation is necessary to gain a deeper understanding of the mechanisms of action and potential side effects of silver nanoparticles on HUVEC cells.

摘要索非拉是一种民族药用植物，含有抗氧化成分，可保护细胞完整性免受各种形式的损伤，并可能在癌症预防中发挥作用。这种植物中含有的抗氧化化合物能促进身体产生新细胞，降低患结肠癌的风险。近年来，利用民族药材通过绿色方法合成的纳米银已被用于癌症治疗。我们研究了采用绿色化学原理合成的银纳米颗粒，利用鸢尾叶提取物治疗肺癌。评估了Ag NPs对流行肺癌细胞的作用。利用x射线衍射（XRD）、场发射扫描电子显微镜（FE-SEM）、紫外可见光谱（UV-Vis）、能量色散x射线光谱（EDX）和透射电子显微镜（TEM）对绿色合成的纳米银进行了表征。形态学分析的结果证实了纳米颗粒的球形，其尺寸范围从20到60纳米。HLC-1、LC-2/ad和PC-14细胞株的IC50值分别为173、125和109 μg/mL。根据最近的数据，Ag NPs可能是支持肺癌治疗的有用选择。虽然目前的研究结果令人鼓舞，但需要进一步的研究来更深入地了解银纳米颗粒对HUVEC细胞的作用机制和潜在的副作用。

{"title":"Targeting the HLC-1, LC-2/ad, and PC-14 lung cancer cell lines by the silver nanoparticles green-formulated by Descurainia sophia leaf extract","authors":"Jianjun Ge, Jianbo Wen, Mingjun Jiang, Kefeng Huang, Saichun Qi, Wei Huang, Linlin Tan","doi":"10.1016/j.mcp.2024.102001","DOIUrl":"10.1016/j.mcp.2024.102001","url":null,"abstract":"<div><div><em>Descurainia sophia</em>, as an an ethno-medicinal plant, contains antioxidant compounds that safeguard cellular integrity against various forms of damage and may play a role in cancer prevention. Antioxidant compounds present in this plant facilitate the body's production of new cells and diminish the risk of colon cancer. In recent years, silver nanoparticles synthesized through green methods using ethnomedicinal herbs have been employed in cancers treatment. We have conducted an investigation into silver nanoparticles that were synthesized through green chemistry principles, utilizing the <em>Descurainia sophia</em> leaves extract for lung carcinoma treatment. The efficacy of Ag NPs against prevalent lung cancer cells was assessed. The green-synthesized silver nanoparticles characterization was conducted utilizing X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), ultraviolet–visible spectroscopy (UV–Vis), energy-dispersive X-ray spectroscopy (EDX), and transmission electron microscopy (TEM). The findings from morphological analyses validate the nanoparticles spherical shape, which ranges in size from 20 to 60 nm. The IC<sub>50</sub> values were determined to be 173, 125, and 109 μg/mL for HLC-1, LC-2/ad, and PC-14 cell lines, respectively. According to recent data, Ag NPs may be a useful option to support the treatment of lung cancer. Although the current study presents encouraging findings, further investigation is necessary to gain a deeper understanding of the mechanisms of action and potential side effects of silver nanoparticles on HUVEC cells.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 102001"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142830608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

miR-375-3p predicts the severity of endometriosis and regulates cellular progression by targeting NOX4 miR-375-3p通过靶向NOX4预测子宫内膜异位症的严重程度并调节细胞进展。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2024-12-19 DOI: 10.1016/j.mcp.2024.101999

Junmei Wang , Jianling Li , Hua Han , Changhua Wang , Taiying Shi , Xueyun Yang

Background

Due to the complex pathogenesis of endometriosis, its early screening and development prediction are still challenging problems in the clinic.

Objectives

This study evaluated the significance of miR-375-3p in endometriosis onset, progression, and recurrence, aiming to identify a novel biomarker for disease diagnosis and prognosis.

Materials and methods

The study enrolled 100 patients with endometriosis and 80 healthy females. The serum miR-375-3p levels were compared between the two groups, and its diagnostic significance and predictive value were assessed by ROC and Cox regression analyses. The effect of miR-375-3p on endometriosis cell growth and motility was evaluated by CCK8 and Transwell assays.

Results

Endometriosis patients showed a lower serum miR-375-3p level relative to healthy females, and more severe the disease condition, lower the miR-375-3p in endometrial tissues is. Reducing serum miR-375-3p could discriminate endometriosis patients sensitively and specifically. Additionally, miR-375-3p was identified as a predictor for the recurrence of endometriosis together with stage, lesion size, and the levels of related hormones. In endometriosis cells, miR-375-3p was demonstrated to target NOX4 and negatively regulated its expression. Overexpressing miR-375-3p significantly suppressed cell proliferation, migration, and invasion, which was reversed by NOX4.

Conclusion

Decreasing miR-375-3p served as a biomarker for endometriosis onset, development, and recurrence. miR-375-3p regulated endometriosis cell growth and motility via negatively modulating NOX4.

背景：由于子宫内膜异位症的发病机制复杂，其早期筛查和发展预测仍是临床上的难题：由于子宫内膜异位症的发病机制复杂，其早期筛查和发展预测仍是临床上的难题：本研究评估了miR-375-3p在子宫内膜异位症发病、进展和复发中的意义，旨在为疾病诊断和预后确定一种新的生物标志物：研究对象包括100名子宫内膜异位症患者和80名健康女性。比较两组患者的血清 miR-375-3p 水平，并通过 ROC 和 Cox 回归分析评估其诊断意义和预测价值。通过CCK8和Transwell试验评估了miR-375-3p对子宫内膜异位症细胞生长和活力的影响：结果：与健康女性相比，子宫内膜异位症患者的血清 miR-375-3p 水平较低；病情越严重，子宫内膜组织中的 miR-375-3p 水平越低。降低血清中的 miR-375-3p 可以灵敏、特异地鉴别子宫内膜异位症患者。此外，miR-375-3p 与子宫内膜异位症的分期、病灶大小和相关激素水平一起被认为是子宫内膜异位症复发的预测因子。在子宫内膜异位症细胞中，miR-375-3p 被证明靶向 NOX4 并负向调节其表达。过表达 miR-375-3p 能显著抑制细胞增殖、迁移和侵袭，而 NOX4 能逆转这种抑制作用：miR-375-3p通过负向调节NOX4来调控子宫内膜异位症细胞的生长和运动。

{"title":"miR-375-3p predicts the severity of endometriosis and regulates cellular progression by targeting NOX4","authors":"Junmei Wang , Jianling Li , Hua Han , Changhua Wang , Taiying Shi , Xueyun Yang","doi":"10.1016/j.mcp.2024.101999","DOIUrl":"10.1016/j.mcp.2024.101999","url":null,"abstract":"<div><h3>Background</h3><div>Due to the complex pathogenesis of endometriosis, its early screening and development prediction are still challenging problems in the clinic.</div></div><div><h3>Objectives</h3><div>This study evaluated the significance of miR-375-3p in endometriosis onset, progression, and recurrence, aiming to identify a novel biomarker for disease diagnosis and prognosis.</div></div><div><h3>Materials and methods</h3><div>The study enrolled 100 patients with endometriosis and 80 healthy females. The serum miR-375-3p levels were compared between the two groups, and its diagnostic significance and predictive value were assessed by ROC and Cox regression analyses. The effect of miR-375-3p on endometriosis cell growth and motility was evaluated by CCK8 and Transwell assays.</div></div><div><h3>Results</h3><div>Endometriosis patients showed a lower serum miR-375-3p level relative to healthy females, and more severe the disease condition, lower the miR-375-3p in endometrial tissues is. Reducing serum miR-375-3p could discriminate endometriosis patients sensitively and specifically. Additionally, miR-375-3p was identified as a predictor for the recurrence of endometriosis together with stage, lesion size, and the levels of related hormones. In endometriosis cells, miR-375-3p was demonstrated to target NOX4 and negatively regulated its expression. Overexpressing miR-375-3p significantly suppressed cell proliferation, migration, and invasion, which was reversed by NOX4.</div></div><div><h3>Conclusion</h3><div>Decreasing miR-375-3p served as a biomarker for endometriosis onset, development, and recurrence. miR-375-3p regulated endometriosis cell growth and motility via negatively modulating NOX4.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 101999"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Circulating miR-574–5p shows diagnostic and prognostic significance and regulates oxygen-glucose deprivation (OGD)-induced inflammatory activation of microglia by targeting ATP2B2 循环miR-574-5p具有诊断和预后意义，并通过靶向ATP2B2调节氧葡萄糖剥夺（OGD）诱导的小胶质细胞炎症激活。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2025-02-08 DOI: 10.1016/j.mcp.2025.102016

Xia Yin , Chunlei Zhang

Background

Early screening is critical for the prevention of ischemic stroke. miR-574–5p was considered a promising biomarker for ischemic stroke but lacks direct confirmation. This study evaluated miR-574–5p in discriminating ischemic stroke and predicting the severity and prognosis of patients, aiming to provide novel insights into the clinical prevention of ischemic stroke.

Methods

The clinical significance of miR-574–5p was evaluated in 103 ischemic stroke patients with 87 healthy individuals as control. The potential of serum miR-574–5p in the diagnosis and prognosis of ischemic stroke was assessed by ROC and logistic regression analyses. In vitro, oxygen-glucose deprivation (OGD)-induced microglia was established. The regulation of inflammation, oxidative stress, and proliferation of microglia by miR-574–5p were assessed by cell transfection. The downstream targets of miR-574–5p were predicted from public databases, and the targeting relationship was evaluated by luciferase reporter assay.

Results

Reducing serum miR-574–5p was observed in ischemic stroke patients relative to healthy individuals, which discriminated ischemic stroke patients. Serum miR-574–5p was negatively correlated with the NIHSS score of ischemic stroke patients and was identified as a risk factor for patients’ adverse prognosis. In OGD-induced microglia, overexpressing miR-574–5p could alleviate OGD-induced inflammation and oxidative stress and promote cell growth. Among predicted targets, ATP2B2 was upregulated in ischemic stroke and showed a negative correlation with miR-574–5p. miR-574–5p negatively regulated ATP2B2 in OGD-induced microglia, and the overexpression of ATP2B2 reversed the protective effect of miR-574–5p.

Conclusion

miR-574–5p acted as a biomarker for ischemic stroke and mediated neuroinflammation via targeting ATP2B2.

背景：早期筛查对预防缺血性脑卒中至关重要。miR-574-5p被认为是缺血性卒中的有希望的生物标志物，但缺乏直接证实。本研究评估miR-574-5p在判别缺血性脑卒中、预测患者严重程度和预后中的作用，旨在为缺血性脑卒中的临床预防提供新的见解。方法：评价103例缺血性脑卒中患者miR-574-5p的临床意义，87例健康人作为对照。采用ROC和logistic回归分析评估血清miR-574-5p在缺血性脑卒中诊断和预后中的潜力。体外建立氧葡萄糖剥夺（OGD）诱导的小胶质细胞。通过细胞转染评估miR-574-5p对炎症、氧化应激和小胶质细胞增殖的调节作用。从公共数据库中预测miR-574-5p的下游靶标，并通过荧光素酶报告基因检测评估其靶向关系。结果：缺血性脑卒中患者血清miR-574-5p较健康人降低，可区分缺血性脑卒中患者。血清miR-574-5p与缺血性脑卒中患者NIHSS评分呈负相关，被认为是患者预后不良的危险因素。在ogd诱导的小胶质细胞中，过表达miR-574-5p可以减轻ogd诱导的炎症和氧化应激，促进细胞生长。在预测的靶点中，ATP2B2在缺血性卒中中上调，并与miR-574-5p呈负相关。在ogd诱导的小胶质细胞中，miR-574-5p负向调节ATP2B2， ATP2B2的过表达逆转了miR-574-5p的保护作用。结论：miR-574-5p通过靶向ATP2B2作为缺血性卒中和介导的神经炎症的生物标志物。

{"title":"Circulating miR-574–5p shows diagnostic and prognostic significance and regulates oxygen-glucose deprivation (OGD)-induced inflammatory activation of microglia by targeting ATP2B2","authors":"Xia Yin , Chunlei Zhang","doi":"10.1016/j.mcp.2025.102016","DOIUrl":"10.1016/j.mcp.2025.102016","url":null,"abstract":"<div><h3>Background</h3><div>Early screening is critical for the prevention of ischemic stroke. miR-574–5p was considered a promising biomarker for ischemic stroke but lacks direct confirmation. This study evaluated miR-574–5p in discriminating ischemic stroke and predicting the severity and prognosis of patients, aiming to provide novel insights into the clinical prevention of ischemic stroke.</div></div><div><h3>Methods</h3><div>The clinical significance of miR-574–5p was evaluated in 103 ischemic stroke patients with 87 healthy individuals as control. The potential of serum miR-574–5p in the diagnosis and prognosis of ischemic stroke was assessed by ROC and logistic regression analyses. In vitro, oxygen-glucose deprivation (OGD)-induced microglia was established. The regulation of inflammation, oxidative stress, and proliferation of microglia by miR-574–5p were assessed by cell transfection. The downstream targets of miR-574–5p were predicted from public databases, and the targeting relationship was evaluated by luciferase reporter assay.</div></div><div><h3>Results</h3><div>Reducing serum miR-574–5p was observed in ischemic stroke patients relative to healthy individuals, which discriminated ischemic stroke patients. Serum miR-574–5p was negatively correlated with the NIHSS score of ischemic stroke patients and was identified as a risk factor for patients’ adverse prognosis. In OGD-induced microglia, overexpressing miR-574–5p could alleviate OGD-induced inflammation and oxidative stress and promote cell growth. Among predicted targets, ATP2B2 was upregulated in ischemic stroke and showed a negative correlation with miR-574–5p. miR-574–5p negatively regulated ATP2B2 in OGD-induced microglia, and the overexpression of ATP2B2 reversed the protective effect of miR-574–5p.</div></div><div><h3>Conclusion</h3><div>miR-574–5p acted as a biomarker for ischemic stroke and mediated neuroinflammation via targeting ATP2B2.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 102016"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143069239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

miR-28–3p suppresses gastric cancer growth and EMT-driven metastasis by targeting the ARF6/Hedgehog axis miR-28-3p通过靶向ARF6/Hedgehog轴抑制胃癌生长和emt驱动的转移。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2025-02-01 Epub Date: 2025-01-09 DOI: 10.1016/j.mcp.2025.102010

Hua Ji , Sicheng Liu , Libo Yang , Yunhua Wu , Huanqing Zhang , Xueqing Liu , Linhai Li , Lihua Li

Gastric cancer (GC), among the most prevalent malignant tumors globally, demonstrates a rapid metastasis rate leading to high mortality. While microRNAs (miRNAs) have been recognized as critical regulators of tumor progression, the specific role of miR-28–3p in GC remains unclear. In this study, we demonstrate that miR-28–3p acts as a tumor suppressor by inhibiting GC cell proliferation and EMT-driven migration in vitro, as well as tumor growth and metastasis in vivo. Mechanistically, miR-28–3p directly targets ADP ribosylation factor 6 (ARF6), a small GTPase identified as an oncogene in GC. We reveal that ARF6 is significantly upregulated in GC and activates the GLI1/2-dependent Hedgehog signaling pathway, promoting tumor growth and EMT. Notably, ARF6 knockdown mitigates the pro-tumor effects caused by miR-28–3p deficiency, while combined ARF6 inhibition and Hedgehog pathway suppression exhibit synergistic anti-tumor effects. This study establishes the miR-28-3p-ARF6-Hedgehog signaling axis as a critical regulatory pathway in GC progression. Our findings provide novel insights into GC pathogenesis and highlight the therapeutic potential of targeting this axis for innovative treatment strategies.

胃癌（GC）是全球最常见的恶性肿瘤之一，其转移率快，死亡率高。虽然microRNAs （miRNAs）已被认为是肿瘤进展的关键调节因子，但miR-28-3p在胃癌中的具体作用尚不清楚。在本研究中，我们证明了miR-28-3p在体外通过抑制GC细胞增殖和emt驱动的迁移以及体内肿瘤生长和转移来发挥肿瘤抑制作用。在机制上，miR-28-3p直接靶向ADP核糖化因子6 (ARF6)，这是一种小的GTPase，在GC中被认为是一种致癌基因。我们发现ARF6在GC中显著上调，激活gli1 /2依赖性Hedgehog信号通路，促进肿瘤生长和EMT。值得注意的是，ARF6敲低可减轻miR-28-3p缺失引起的促肿瘤作用，而ARF6抑制与Hedgehog途径抑制联合作用则具有协同抗肿瘤作用。本研究确定miR-28-3p-ARF6-Hedgehog信号轴是GC进展的关键调控途径。我们的研究结果为GC的发病机制提供了新的见解，并强调了针对这一轴的创新治疗策略的治疗潜力。

{"title":"miR-28–3p suppresses gastric cancer growth and EMT-driven metastasis by targeting the ARF6/Hedgehog axis","authors":"Hua Ji , Sicheng Liu , Libo Yang , Yunhua Wu , Huanqing Zhang , Xueqing Liu , Linhai Li , Lihua Li","doi":"10.1016/j.mcp.2025.102010","DOIUrl":"10.1016/j.mcp.2025.102010","url":null,"abstract":"<div><div>Gastric cancer (GC), among the most prevalent malignant tumors globally, demonstrates a rapid metastasis rate leading to high mortality. While microRNAs (miRNAs) have been recognized as critical regulators of tumor progression, the specific role of miR-28–3p in GC remains unclear. In this study, we demonstrate that miR-28–3p acts as a tumor suppressor by inhibiting GC cell proliferation and EMT-driven migration in vitro, as well as tumor growth and metastasis in vivo. Mechanistically, miR-28–3p directly targets ADP ribosylation factor 6 (ARF6), a small GTPase identified as an oncogene in GC. We reveal that ARF6 is significantly upregulated in GC and activates the GLI1/2-dependent Hedgehog signaling pathway, promoting tumor growth and EMT. Notably, ARF6 knockdown mitigates the pro-tumor effects caused by miR-28–3p deficiency, while combined ARF6 inhibition and Hedgehog pathway suppression exhibit synergistic anti-tumor effects. This study establishes the miR-28-3p-ARF6-Hedgehog signaling axis as a critical regulatory pathway in GC progression. Our findings provide novel insights into GC pathogenesis and highlight the therapeutic potential of targeting this axis for innovative treatment strategies.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"79 ","pages":"Article 102010"},"PeriodicalIF":2.3,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142957845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage 敲除 Caspase-1 可破坏裂解过程并保护感光细胞免受光化学损伤

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2024-12-01 Epub Date: 2024-11-17 DOI: 10.1016/j.mcp.2024.101991

Xiaoping Yu , Jiayuan Peng , Qian Zhong , Ailin Wu , Xiaoming Deng , Yanfeng Zhu

Aim

Retinal photochemical damage (RPD) plays a significant role in the development of various ocular diseases, with Caspase-1 being a key contributor. This study investigates the protective effects of Caspase-1 gene-mediated pyroptosis against RPD.

Methods

Differentially expressed genes (DEGs) associated with RPD were identified through the analysis of two expression profiles from the GEO database. Correlation analysis was used to pinpoint pyroptosis-related genes (PRGs) linked to RPD. A Caspase-1 knockout 661 W cell line was generated via CRISPR-Cas9 gene editing, and single-cell colonies were screened and purified. Validation of knockout cells was performed through RT-qPCR, gene sequencing, and Western blot analysis. Comparative assays on cell proliferation, intracellular reactive oxygen species (ROS), and cytotoxicity were conducted between wild-type and Caspase-1 knockout cells under light exposure. Further RT-qPCR and Western blot experiments examined changes in the mRNA and protein levels of key pyroptosis pathway components.

Results

Significant alterations in Caspase-1 expression were observed among PRGs. Homozygous Caspase-1 knockout cell lines were confirmed through RT-qPCR, genomic PCR product sequencing, and Western blot analysis. Compared to wild-type 661 W cells, Caspase-1 knockout cells exhibited higher viability and proliferation rates after 24 h of light exposure, alongside reduced LDH release. The expression of downstream pyroptosis factors at both the mRNA and protein levels was markedly decreased in the knockout group.

Conclusion

CRISPR/Cas9-mediated Caspase-1 knockout enhanced the resistance of 661 W cells to photochemical damage, suggesting that Caspase-1 may serve as a potential therapeutic target for RPD-related diseases.

目的：视网膜光化学损伤（RPD）在各种眼部疾病的发生发展中起着重要作用，而Caspase-1是其中的关键因素。本研究探讨了 Caspase-1 基因介导的热蛋白沉积对 RPD 的保护作用：方法：通过分析 GEO 数据库中的两个表达谱，确定了与 RPD 相关的差异表达基因（DEGs）。方法：通过分析 GEO 数据库中的两个表达谱，确定了与 RPD 相关的差异表达基因（DEGs），并利用相关性分析确定了与 RPD 相关的热蛋白沉积相关基因（PRGs）。通过 CRISPR-Cas9 基因编辑生成了 Caspase-1 基因敲除的 661W 细胞系，并筛选和纯化了单细胞菌落。通过 RT-qPCR、基因测序和 Western 印迹分析对基因敲除细胞进行了验证。在光照条件下，对野生型细胞和 Caspase-1 基因敲除细胞的细胞增殖、细胞内活性氧（ROS）和细胞毒性进行了比较分析。进一步的 RT-qPCR 和 Western 印迹实验检测了主要热蛋白沉积途径成分的 mRNA 和蛋白质水平的变化：结果：在 PRGs 中观察到 Caspase-1 表达的显著变化。通过 RT-qPCR、基因组 PCR 产物测序和 Western 印迹分析确认了同基因 Caspase-1 基因敲除细胞系。与野生型 661W 细胞相比，Caspase-1 基因敲除细胞在光照 24 小时后表现出更高的存活率和增殖率，同时 LDH 释放量减少。Caspase-1基因敲除组的下游热休克因子在mRNA和蛋白质水平上的表达都明显下降：结论：CRISPR/Cas9介导的Caspase-1基因敲除增强了661W细胞对光化学损伤的抵抗力，这表明Caspase-1可作为RPD相关疾病的潜在治疗靶点。

{"title":"Caspase-1 knockout disrupts pyroptosis and protects photoreceptor cells from photochemical damage","authors":"Xiaoping Yu , Jiayuan Peng , Qian Zhong , Ailin Wu , Xiaoming Deng , Yanfeng Zhu","doi":"10.1016/j.mcp.2024.101991","DOIUrl":"10.1016/j.mcp.2024.101991","url":null,"abstract":"<div><h3>Aim</h3><div>Retinal photochemical damage (RPD) plays a significant role in the development of various ocular diseases, with Caspase-1 being a key contributor. This study investigates the protective effects of Caspase-1 gene-mediated pyroptosis against RPD.</div></div><div><h3>Methods</h3><div>Differentially expressed genes (DEGs) associated with RPD were identified through the analysis of two expression profiles from the GEO database. Correlation analysis was used to pinpoint pyroptosis-related genes (PRGs) linked to RPD. A Caspase-1 knockout 661 W cell line was generated via CRISPR-Cas9 gene editing, and single-cell colonies were screened and purified. Validation of knockout cells was performed through RT-qPCR, gene sequencing, and Western blot analysis. Comparative assays on cell proliferation, intracellular reactive oxygen species (ROS), and cytotoxicity were conducted between wild-type and Caspase-1 knockout cells under light exposure. Further RT-qPCR and Western blot experiments examined changes in the mRNA and protein levels of key pyroptosis pathway components.</div></div><div><h3>Results</h3><div>Significant alterations in Caspase-1 expression were observed among PRGs. Homozygous Caspase-1 knockout cell lines were confirmed through RT-qPCR, genomic PCR product sequencing, and Western blot analysis. Compared to wild-type 661 W cells, Caspase-1 knockout cells exhibited higher viability and proliferation rates after 24 h of light exposure, alongside reduced LDH release. The expression of downstream pyroptosis factors at both the mRNA and protein levels was markedly decreased in the knockout group.</div></div><div><h3>Conclusion</h3><div>CRISPR/Cas9-mediated Caspase-1 knockout enhanced the resistance of 661 W cells to photochemical damage, suggesting that Caspase-1 may serve as a potential therapeutic target for RPD-related diseases.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"78 ","pages":"Article 101991"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142590893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Prognostic prediction of gastric cancer based on H&E findings and machine learning pathomics 基于 H&E 检查结果和机器学习病理组学的胃癌预后预测。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2024-12-01 Epub Date: 2024-09-30 DOI: 10.1016/j.mcp.2024.101983

Guoda Han , Xu Liu , Tian Gao , Lei Zhang , Xiaoling Zhang , Xiaonan Wei , Yecheng Lin , Bohong Yin

Aim

In this research, we aimed to develop a model for the accurate prediction of gastric cancer based on H&E findings combined with machine learning pathomics.

Methods

Transcriptome data, pathological images, and clinical data from 443 cases were retrieved from TCGA (The Cancer Genome Atlas Program) for survival analysis. The images were segmented using the Otsu algorithm, and features were extracted using the PyRadiomics package. Subsequently, the cases were randomly divided into a training cohort of 165 cases and a validation cohort of 69 cases. Features selected via minimum Redundancy - Maximum Relevance (mRMR)- recursive feature elimination (RFE) screening were used to train a model using the Gradient Boosting Machine (GBM) algorithm. The model's performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), calibration curves, and decision curves. Additionally, the correlation between the Pathomics score (PS) and immune genes was examined.

Results

In the multivariate analysis, heightened infiltration of activated CD4 memory T cells was strongly associated with improved overall survival (HR = 0.505, 95 % CI = 0.342–0.745, P < 0.001). The pathomic model, exhibiting robust predictive capability, demonstrated impressive AUC values of 0.844 and 0.750 in both study cohorts. The Decision Curve Analysis (DCA) unequivocally underscored the model's exceptional clinical utility. In a subsequent multivariate analysis, heightened infiltration of the PS also emerged as a significant protective factor for overall survival (HR = 0.506, 95 % CI = 0.329–0.777, P = 0.002).

Conclusion

The pathomic model based on H&E slides for predicting the infiltration degree of activated CD4 memory T cells, along with integrated bioinformatics analysis elucidating potential molecular mechanisms, offers novel prognostic indicators for the precise stratification and individualized prognosis of gastric cancer patients.

目的：在这项研究中，我们旨在开发一种基于H&E结果并结合机器学习病理组学的胃癌准确预测模型：方法：我们从 TCGA（癌症基因组图谱计划）中获取了 443 个病例的转录组数据、病理图像和临床数据，用于生存分析。使用Otsu算法对图像进行分割，并使用PyRadiomics软件包提取特征。随后，病例被随机分为 165 例训练队列和 69 例验证队列。通过最小冗余-最大相关性（mRMR）-递归特征剔除（RFE）筛选出的特征被用于使用梯度提升机（GBM）算法训练模型。使用接收者操作特征曲线（ROC）下面积（AUC）、校准曲线和决策曲线对模型的性能进行了评估。此外，还研究了病理组学评分（PS）与免疫基因之间的相关性：在多变量分析中，活化的 CD4 记忆 T 细胞浸润增加与总生存期的改善密切相关（HR = 0.505，95% CI = 0.342-0.745，P <0.001）。病理模型具有强大的预测能力，在两个研究队列中的AUC值分别为0.844和0.750，令人印象深刻。决策曲线分析（DCA）明确强调了该模型卓越的临床实用性。在随后的多变量分析中，PS的高度浸润也成为总生存率的重要保护因素（HR = 0.506，95% CI = 0.329-0.777，P = 0.002）：基于H&E切片预测活化CD4记忆T细胞浸润程度的病理模型，以及阐明潜在分子机制的综合生物信息学分析，为胃癌患者的精确分层和个体化预后提供了新的预后指标。

{"title":"Prognostic prediction of gastric cancer based on H&E findings and machine learning pathomics","authors":"Guoda Han , Xu Liu , Tian Gao , Lei Zhang , Xiaoling Zhang , Xiaonan Wei , Yecheng Lin , Bohong Yin","doi":"10.1016/j.mcp.2024.101983","DOIUrl":"10.1016/j.mcp.2024.101983","url":null,"abstract":"<div><h3>Aim</h3><div>In this research, we aimed to develop a model for the accurate prediction of gastric cancer based on H&E findings combined with machine learning pathomics.</div></div><div><h3>Methods</h3><div>Transcriptome data, pathological images, and clinical data from 443 cases were retrieved from TCGA (The Cancer Genome Atlas Program) for survival analysis. The images were segmented using the Otsu algorithm, and features were extracted using the PyRadiomics package. Subsequently, the cases were randomly divided into a training cohort of 165 cases and a validation cohort of 69 cases. Features selected via minimum Redundancy - Maximum Relevance (mRMR)- recursive feature elimination (RFE) screening were used to train a model using the Gradient Boosting Machine (GBM) algorithm. The model's performance was evaluated using the area under the receiver operating characteristic (ROC) curve (AUC), calibration curves, and decision curves. Additionally, the correlation between the Pathomics score (PS) and immune genes was examined.</div></div><div><h3>Results</h3><div>In the multivariate analysis, heightened infiltration of activated CD4 memory T cells was strongly associated with improved overall survival (HR = 0.505, 95 % CI = 0.342–0.745, P < 0.001). The pathomic model, exhibiting robust predictive capability, demonstrated impressive AUC values of 0.844 and 0.750 in both study cohorts. The Decision Curve Analysis (DCA) unequivocally underscored the model's exceptional clinical utility. In a subsequent multivariate analysis, heightened infiltration of the PS also emerged as a significant protective factor for overall survival (HR = 0.506, 95 % CI = 0.329–0.777, P = 0.002).</div></div><div><h3>Conclusion</h3><div>The pathomic model based on H&E slides for predicting the infiltration degree of activated CD4 memory T cells, along with integrated bioinformatics analysis elucidating potential molecular mechanisms, offers novel prognostic indicators for the precise stratification and individualized prognosis of gastric cancer patients.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"78 ","pages":"Article 101983"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142299499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Serum miR-155–5p assists the diagnostic sensitivity and accuracy of low-dose spiral CT imaging in early lung cancer 血清miR-155-5p有助于早期肺癌低剂量螺旋CT成像诊断的敏感性和准确性。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2024-12-01 Epub Date: 2024-12-04 DOI: 10.1016/j.mcp.2024.101994

Fei Li, Wenwen Han, Hailong Sun

Background

Low-dose spiral CT imaging has been employed for cancer diagnosis, but its sensitivity and specificity were unsatisfactory. MicroRNAs have been considered an approach for screening cancers, and the function of miR-155–5p in lung cancer has been previously revealed.

Objectives

This study assessed the diagnostic value of combining low-dose spiral CT with serum miR-155–5p in lung cancer aiming to explore a novel strategy to assist the clinical cancer diagnosis.

Methods

This study enrolled 115 lung cancer patients and 115 patients with benign lung diseases as control. All patients received low-dose spiral CT imaging, and serum miR-155–5p levels were analyzed by PCR. The diagnostic potential of miR-155–5p in lung cancer was evaluated by receiver operating curve (ROC), and its significance in evaluating the risk of lung cancer was evaluated by logistic regression analysis. The consistency of serum miR-155–5p and low-dose spiral CT imaging with pathological examination was assessed by the Kappa test.

Results

Reduced serum miR-155–5p indicated the risk of lung cancer in patients with benign lung diseases. Decreased serum miR-155–5p showed significant diagnostic value in early lung cancer and was significantly associated with disease severity. Low-dose spiral CT imaging showed significant diagnostic value in early lung cancer and showed middle consistency with pathological examination. Combining low-dose spiral CT imaging with serum miR-155–5p improved the diagnostic sensitivity and accuracy in early lung cancer, reduced the false positive rate, and showed better consistency with pathological examination.

Conclusion

Serum miR-155–5p levels could assist the early detection of lung cancer by low-dose spiral CT examination.

背景：低剂量螺旋CT成像已被用于肿瘤诊断，但其敏感性和特异性不理想。microrna一直被认为是筛查癌症的一种方法，miR-155-5p在肺癌中的功能此前已被揭示。目的：本研究评估低剂量螺旋CT联合血清miR-155-5p对肺癌的诊断价值，旨在探索辅助临床癌症诊断的新策略。方法：本研究纳入115例肺癌患者和115例肺部良性疾病患者作为对照。所有患者均接受低剂量螺旋CT成像，并通过PCR分析血清miR-155-5p水平。采用受试者工作曲线（receiver operating curve， ROC）评价miR-155-5p在肺癌中的诊断潜力，采用logistic回归分析评价miR-155-5p在肺癌风险评价中的意义。采用Kappa试验评估血清miR-155-5p及低剂量螺旋CT成像与病理检查的一致性。结果：血清miR-155-5p降低提示肺部良性疾病患者发生肺癌的风险。血清miR-155-5p降低对早期肺癌具有重要的诊断价值，且与疾病严重程度显著相关。低剂量螺旋CT对早期肺癌的诊断价值显著，与病理检查的一致性中等。低剂量螺旋CT影像联合血清miR-155-5p可提高早期肺癌诊断的敏感性和准确性，降低假阳性率，与病理检查的一致性更好。结论：血清miR-155-5p水平有助于低剂量螺旋CT检查早期发现肺癌。

{"title":"Serum miR-155–5p assists the diagnostic sensitivity and accuracy of low-dose spiral CT imaging in early lung cancer","authors":"Fei Li, Wenwen Han, Hailong Sun","doi":"10.1016/j.mcp.2024.101994","DOIUrl":"10.1016/j.mcp.2024.101994","url":null,"abstract":"<div><h3>Background</h3><div>Low-dose spiral CT imaging has been employed for cancer diagnosis, but its sensitivity and specificity were unsatisfactory. MicroRNAs have been considered an approach for screening cancers, and the function of miR-155–5p in lung cancer has been previously revealed.</div></div><div><h3>Objectives</h3><div>This study assessed the diagnostic value of combining low-dose spiral CT with serum miR-155–5p in lung cancer aiming to explore a novel strategy to assist the clinical cancer diagnosis.</div></div><div><h3>Methods</h3><div>This study enrolled 115 lung cancer patients and 115 patients with benign lung diseases as control. All patients received low-dose spiral CT imaging, and serum miR-155–5p levels were analyzed by PCR. The diagnostic potential of miR-155–5p in lung cancer was evaluated by receiver operating curve (ROC), and its significance in evaluating the risk of lung cancer was evaluated by logistic regression analysis. The consistency of serum miR-155–5p and low-dose spiral CT imaging with pathological examination was assessed by the Kappa test.</div></div><div><h3>Results</h3><div>Reduced serum miR-155–5p indicated the risk of lung cancer in patients with benign lung diseases. Decreased serum miR-155–5p showed significant diagnostic value in early lung cancer and was significantly associated with disease severity. Low-dose spiral CT imaging showed significant diagnostic value in early lung cancer and showed middle consistency with pathological examination. Combining low-dose spiral CT imaging with serum miR-155–5p improved the diagnostic sensitivity and accuracy in early lung cancer, reduced the false positive rate, and showed better consistency with pathological examination.</div></div><div><h3>Conclusion</h3><div>Serum miR-155–5p levels could assist the early detection of lung cancer by low-dose spiral CT examination.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"78 ","pages":"Article 101994"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142755758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

LncRNA PVT1 promotes malignant progression by regulating the miR-7-5p/CDKL1 axis in oral squamous cell carcinoma LncRNA PVT1通过调节miR-7-5p/CDKL1轴在口腔鳞状细胞癌中促进恶性进展。

IF 2.3 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Molecular and Cellular Probes

Pub Date : 2024-12-01 Epub Date: 2024-12-02 DOI: 10.1016/j.mcp.2024.101995

Jun Li , Shuxin Ding , Min Li , Bo Zou , Miaomiao Chu , Guohao Gu , Cheng Chen , Yu-jiao Liu , Ke Zheng , Zhen Meng

Oral squamous cell carcinoma (OSCC), one of the most common types of head and neck squamous cell carcinoma (HNSCC), is characterized by high incidence and mortality. PVT1 is a long non-coding RNA (lncRNA) that plays an oncogenic role in various cancer types. This study aims to reveal the role and underlying molecular mechanism of PVT1 in OSCC progression. The expression levels of PVT1, miR-7-5p, and CDKL1 mRNA were evaluated using qRT-PCR. Western blot and IHC analysis were conducted to determine the protein expression of CDKL1. The biological functions of PVT1, miR-7-5p, and CDKL1 in OSCC were investigated through CCK-8, transwell migration and invasion assays. In vivo experiments utilized a xenograft model to examine the impact of PVT1 on OSCC. Furthermore, the interaction among PVT1, miR-7-5p, and CDKL1 was explored using RNA pull down assay and luciferase reporter assays. We found that PVT1 enhanced cell proliferation, migration, and invasion by targeting CDKL1. In addition, PVT1 functions as a sponge to modulate miR-7-5p, thereby influencing the expression of CDKL1 and the progression of OSCC. In conclusion, this study illustrates that the "PVT1/miR-7-5p/CDKL1" pathway is capable of promoting the progression of OSCC and may serve as a promising target for developing treatment strategies for OSCC.

口腔鳞状细胞癌（Oral squamous cell carcinoma， OSCC）是头颈部鳞状细胞癌（HNSCC）中最常见的类型之一，具有高发病率和高死亡率的特点。PVT1是一种长链非编码RNA (lncRNA)，在多种癌症类型中发挥致癌作用。本研究旨在揭示PVT1在OSCC进展中的作用及其分子机制。采用qRT-PCR技术评估PVT1、miR-7-5p和CDKL1 mRNA的表达水平。Western blot和IHC检测CDKL1蛋白表达。通过CCK-8、跨井迁移和侵袭试验研究PVT1、miR-7-5p和CDKL1在OSCC中的生物学功能。体内实验采用异种移植模型来研究PVT1对OSCC的影响。此外，通过RNA拉下实验和荧光素酶报告基因检测，探讨了PVT1、miR-7-5p和CDKL1之间的相互作用。我们发现PVT1通过靶向CDKL1增强细胞增殖、迁移和侵袭。此外，PVT1作为海绵调节miR-7-5p，从而影响CDKL1的表达和OSCC的进展。总之，本研究表明，“PVT1/miR-7-5p/CDKL1”通路能够促进OSCC的进展，并可能作为制定OSCC治疗策略的有希望的靶点。

{"title":"LncRNA PVT1 promotes malignant progression by regulating the miR-7-5p/CDKL1 axis in oral squamous cell carcinoma","authors":"Jun Li , Shuxin Ding , Min Li , Bo Zou , Miaomiao Chu , Guohao Gu , Cheng Chen , Yu-jiao Liu , Ke Zheng , Zhen Meng","doi":"10.1016/j.mcp.2024.101995","DOIUrl":"10.1016/j.mcp.2024.101995","url":null,"abstract":"<div><div>Oral squamous cell carcinoma (OSCC), one of the most common types of head and neck squamous cell carcinoma (HNSCC), is characterized by high incidence and mortality. PVT1 is a long non-coding RNA (lncRNA) that plays an oncogenic role in various cancer types. This study aims to reveal the role and underlying molecular mechanism of PVT1 in OSCC progression. The expression levels of PVT1, miR-7-5p, and CDKL1 mRNA were evaluated using qRT-PCR. Western blot and IHC analysis were conducted to determine the protein expression of CDKL1. The biological functions of PVT1, miR-7-5p, and CDKL1 in OSCC were investigated through CCK-8, transwell migration and invasion assays. In vivo experiments utilized a xenograft model to examine the impact of PVT1 on OSCC. Furthermore, the interaction among PVT1, miR-7-5p, and CDKL1 was explored using RNA pull down assay and luciferase reporter assays. We found that PVT1 enhanced cell proliferation, migration, and invasion by targeting CDKL1. In addition, PVT1 functions as a sponge to modulate miR-7-5p, thereby influencing the expression of CDKL1 and the progression of OSCC. In conclusion, this study illustrates that the \"PVT1/miR-7-5p/CDKL1\" pathway is capable of promoting the progression of OSCC and may serve as a promising target for developing treatment strategies for OSCC.</div></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"78 ","pages":"Article 101995"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142774120","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0