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Melittin Prevents Metastasis of Epidermal Growth Factor-Induced MDA-MB-231 Cells through The Inhibition of The SDF-1α/CXCR4 Signaling Pathway 蜂毒素通过抑制SDF-1α/CXCR4信号通路阻止表皮生长因子诱导的MDA-MB-231细胞转移
Pub Date : 2022-02-01 DOI: 10.22074/cellj.2022.7626
F. Salimian, M. Nabiuni, E. Salehghamari
Objective Melittin is one of the natural components of bee venom (Apis mellifera), and its anticancer and antimetastatic properties have been well established, but the underlying mechanism remains elusive. The MDA-MB-231 is a triple- negative cell line that is highly aggressive and invasive. Besides, many critical proteins are involved in tumor invasion and metastasis. In this study, we investigated whether melittin inhibits the migration and metastasis of epidermal growth factor (EGF)-induced MDA-MB-231 cells via the suppression of SDF-1α/CXCR4 and Rac1-mediated signaling pathways. Materials and Methods In this experimental study, cells were treated with melittin (0.5-4 μg/ml), and the toxicity of melittin was assessed by the MTT assay. Afterward, the migration assay was conducted to measure the degree of the migration of EGF-induced cells. The western blot technique was performed to analyze the rate of Rac1, p-Rac1, SDF- 1α, and CXCR4 expression in different groups. Results The results demonstrated that melittin markedly suppressed the migration of EGF-induced cells and decreased the expression of p-Rac1, CXCR4, and SDF-1α proteins. Conclusion The results of the present study suggested that the anti-tumor properties of melittin could be through the blocking of the SDF-1α/CXCR4 signaling pathway, which is beneficial for the reduction of tumor migration and invasion.
目的蜂毒素是蜂毒的天然成分之一,其抗肿瘤和抗转移特性已被证实,但其作用机制尚不清楚。MDA-MB-231是一种具有高度侵袭性和侵袭性的三阴性细胞系。此外,许多关键蛋白参与了肿瘤的侵袭和转移。在这项研究中,我们研究了蜂毒素是否通过抑制SDF-1α/CXCR4和rac1介导的信号通路来抑制表皮生长因子(EGF)诱导的MDA-MB-231细胞的迁移和转移。材料与方法本实验采用蜂毒素(0.5 ~ 4 μg/ml)处理细胞,采用MTT法评价蜂毒素的毒性。然后进行迁移实验,测定egf诱导细胞的迁移程度。western blot检测各组细胞中Rac1、p-Rac1、SDF- 1α、CXCR4的表达率。结果蜂毒素能明显抑制egf诱导的细胞迁移,降低p-Rac1、CXCR4和SDF-1α蛋白的表达。结论蜂毒素的抗肿瘤作用可能是通过阻断SDF-1α/CXCR4信号通路,有利于减少肿瘤的迁移和侵袭。
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引用次数: 2
Trehalose An Additive Solution for Platelet Concentrate to Protect Platelets from Apoptosis and Clearance during Their Storage at 4°C 海藻糖:血小板浓缩物的添加剂溶液在4°C储存期间保护血小板免于凋亡和清除
Pub Date : 2022-02-01 DOI: 10.22074/cellj.2022.7886
Vahid Baghdadi, R. Ranjbaran, F. Yari, M. Rafiee
Objective Although cold storage of platelets (PLTs) could decrease the risk of bacterial growth, it could affect on the PLTs viability and hemostatic function. At cold temperatures, trehalose can be used to substitute water, inhibit the solid-liquid transition phase of the PLT membrane, and stop Glycoprotein Ibα (GPIbα) polymerization. In this study, we evaluated the potential of trehalose for reducing the negative effects of cold storage on the apoptosis and the clearance rates of PLTs after long-term storage at cold. Materials and Methods In this experimental study, PLT concentrates (PCs) were maintained for five days in the different circumstances. PLTs were subsequently counted by using an automated hematology analyzer. Also water-soluble tetrazolium salt (WST-1) assay was performed to estimate the viability of PLTs. The activity of lactate dehydrogenase enzyme (LDH) was determined by a biochemical analyzer. And human active caspase-3 levels were measured by using enzyme-linked immunosorbent assay (ELISA) method. Also, we applied flow cytometry technique. Results PLTs count and viability were higher, while LDH amount was lower in trehalose-treated PLTs when compared with two other groups (P=0.03). The highest increase in the amount of caspase-3 levels in the PLTs was observed at 4°C. However, trehalose-treated and 4°C PLTs had a lower amount of active caspase-3 in comparison with 4°C PLTs. The level of PS expression on PLTs was lower in the trehalose-treated PLTs in compared with the two other groups (P=0.03). PLTs ingestion by HepG2 cells was enhanced in the 4°C-stored PLTs. However, the ingestion rate was significantly reduced in the trehalose-treated PLTs on day 5 of storage (P=0.03). Conclusion Trehalose can moderate the effects of cold temperature on the apoptosis, viability, and the survival rate of PLTs. It also decreases the ingestion rate of refrigerated PLTs in vitro.
目的冷冻保存血小板虽然能降低细菌滋生的风险,但会影响血小板活力和止血功能。在低温下,海藻糖可以代替水,抑制PLT膜的固液过渡相,阻止糖蛋白Ibα (GPIbα)的聚合。在这项研究中,我们评估了海藻糖在减少冷藏对细胞凋亡的负面影响和长期冷藏后plt清除率方面的潜力。材料与方法在实验研究中,PLT浓缩液(PCs)在不同环境下维持5天。随后使用自动血液学分析仪计数血小板。同时采用水溶性四氮唑盐(WST-1)测定plt的生存能力。用生化分析仪测定乳酸脱氢酶(LDH)活性。采用酶联免疫吸附法(ELISA)检测人活性caspase-3水平。同时应用流式细胞术技术。结果与其他两组相比,海藻糖处理的plt数量和活力更高,LDH含量更低(P=0.03)。在4°C时观察到plt中caspase-3水平的最高增加。然而,海藻糖处理的plt和4°C的plt相比,活性caspase-3的含量较低。海藻糖处理的plt细胞中PS的表达水平低于其他两组(P=0.03)。在4°c储存的plt中,HepG2细胞对plt的摄取增强。然而,海藻糖处理的plt在储存第5天的摄食率显著降低(P=0.03)。结论海藻糖可调节低温对plt细胞凋亡、细胞活力及存活率的影响。它还可以降低体外冷藏plt的摄食率。
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引用次数: 2
Epigenetic Dysregulation of BRDT Gene in Testis Tissues of Infertile Men: Case-Control Study 不育男性睾丸组织BRDT基因的表观遗传失调:病例对照研究
Pub Date : 2022-02-01 DOI: 10.22074/cellj.2022.7724
Fereshteh Kohandani, Parham Jazireian, R. Favaedi, M. S. Sadighi Gilani, S. M. Moshtaghioun, M. Shahhoseini
Objective Bromodomain testis associated (BRDT), a testis-specific member of the Bromo- and Extra-Terrminal domain (BET) protein family, is involved in spermatogenesis and, more specifically, chromatin remodeling. In the post-meiotic spermatogenic cells, BRDT protein binds to the hyperacetylated histones and facilitates their replacement with transition proteins (TPs), particularly protamines, which are essential for chromatin condensation. The current research was conducted to assess the expression and epigenetic profile of BRDT in the testis tissues of infertile men. Materials and Methods In this case-control study, three groups were included: positive control group: obstructive azoospermia (OA, n=10), round spermatid maturation arrest group (SMA, n=10) and negative control group: sertoli cell- only syndrome (SCOS, n=10). Using quantitative real-time polymerase chain reaction (PCR), the expression profile of BRDT was generated. Also, ChIP-real time PCR was used to measure the following histone marks: H3K9ac, H3K9me3, H3K4me3, H3K27me3 on the promoter region of BRDT. Results Our data indicated that BRDT expression decreased in the SMA group in comparison with the positive control group and this finding is in line with the ChIP results obtained in this group. Conclusion Based on these data, we postulate that BRDT gene has a vital role in the spermatogenesis and its decreased expression due to an aberrant epigenetic signaling might be associated with male infertility.
目的溴域睾丸相关蛋白(BRDT)是睾丸特异性的溴域和外端结构域(BET)蛋白家族成员,参与精子发生,更具体地说,参与染色质重塑。在减数分裂后的生精细胞中,BRDT蛋白与高乙酰化的组蛋白结合,并促进它们被过渡蛋白(TPs)取代,特别是对染色质凝聚至关重要的精蛋白。目前的研究旨在评估BRDT在不育男性睾丸组织中的表达和表观遗传谱。材料与方法本病例对照研究分为3组:阳性对照组:阻塞性无精子症(OA, n=10)、圆形精子成熟阻滞组(SMA, n=10)和阴性对照组:仅支持细胞综合征(SCOS, n=10)。采用实时定量聚合酶链反应(PCR),生成BRDT基因的表达谱。采用ChIP-real - time PCR检测BRDT启动子区域的组蛋白标记:H3K9ac、H3K9me3、H3K4me3、H3K27me3。结果我们的数据显示,与阳性对照组相比,SMA组的BRDT表达降低,这一发现与该组的ChIP结果一致。结论BRDT基因在精子发生过程中起重要作用,由于表观遗传信号异常导致BRDT基因表达减少可能与男性不育有关。
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引用次数: 0
Circ_0000228 Promotes Cervical Cancer Progression via Regulating miR-337-3p/TGFBR1 Axis Circ_0000228通过调节miR-337-3p/TGFBR1轴促进宫颈癌进展
Pub Date : 2022-02-01 DOI: 10.22074/cellj.2022.7914
Y. Xu, Xiaona Dong, Baoli Ma, Pingping Mu, Xiang-Heng Kong, Dongmei Li
Objective This study aims to investigate the biological function of circular RNA (circRNA) circ_0000228 in the cervical cancer (CC). Materials and Methods In this experimental study, the GSE113696 dataset was downloaded from the Gene Expression Omnibus (GEO). GEO2R was employed to obtain differentially expressed circRNA between CC tissues and matched paracancerous tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were employed to detect circ_0000228, microRNA-337-3p (miR-337-3p) and transforming growth factor, beta receptor I (TGFBR1) expression levels in the CC tissues and cells. Following gain-of-function and loss-of-function models establishment, CCK-8 and BrdU tests were conducted to examine cell proliferation. Transwell experiment was executed to examine CC cells migration and invasion. A lung metastasis model was utilized to determine the ability of circ_0000228 on the lung metastasis. Bioinformatics analysis, dual-luciferase reporter experiment and RNA immunoprecipitation (RIP) assay were applied to verify the targeting relationship among miR-337-3p, circ_0000228, and TGFBR1. Results Circ_0000228 expression in the CC tissues and cells was up-modulated. Circ_0000228 overexpression markedly enhanced cell proliferation, migration, and invasion, while knocking down circ_0000228 remarkably repressed cell proliferation, migration, and invasion. MiR-337-3p could be adsorbed by circ_0000228. TGFBR1 was identified as a target gene of miR-337-3p that indirectly and positively modulated by circ_0000228 in the CC cells. Conclusion Circ_0000228 up-modulates TGFBR1 by targeting miR-337-3p to enhance CC cell proliferation, migration and invasion. Also, Circ_0000228 is a promising therapeutic target for the CC.
目的探讨环状RNA (circRNA) circ_0000228在宫颈癌(CC)中的生物学功能。材料与方法在本实验研究中,GSE113696数据集从Gene Expression Omnibus (GEO)下载。GEO2R用于获取CC组织和匹配的癌旁组织之间差异表达的circRNA。采用实时荧光定量聚合酶链反应(qRT-PCR)和Western blot检测CC组织和细胞中circ_0000228、microRNA-337-3p (miR-337-3p)和转化生长因子β受体1 (TGFBR1)的表达水平。在建立功能获得和功能丧失模型后,进行CCK-8和BrdU测试以检测细胞增殖。Transwell实验检测CC细胞的迁移和侵袭。利用肺转移模型确定circ_0000228对肺转移的影响。采用生物信息学分析、双荧光素酶报告基因实验和RNA免疫沉淀(RIP)实验验证miR-337-3p、circ_0000228和TGFBR1之间的靶向关系。结果Circ_0000228在CC组织和细胞中的表达上调。过表达Circ_0000228可显著增强细胞增殖、迁移和侵袭,而敲低Circ_0000228可显著抑制细胞增殖、迁移和侵袭。circ_0000228可以吸附MiR-337-3p。TGFBR1被鉴定为miR-337-3p的靶基因,在CC细胞中被circ_0000228间接正调控。结论Circ_0000228通过靶向miR-337-3p上调TGFBR1,促进CC细胞增殖、迁移和侵袭。此外,Circ_0000228是CC的一个有希望的治疗靶点。
{"title":"Circ_0000228 Promotes Cervical Cancer Progression via Regulating miR-337-3p/TGFBR1 Axis","authors":"Y. Xu, Xiaona Dong, Baoli Ma, Pingping Mu, Xiang-Heng Kong, Dongmei Li","doi":"10.22074/cellj.2022.7914","DOIUrl":"https://doi.org/10.22074/cellj.2022.7914","url":null,"abstract":"Objective This study aims to investigate the biological function of circular RNA (circRNA) circ_0000228 in the cervical cancer (CC). Materials and Methods In this experimental study, the GSE113696 dataset was downloaded from the Gene Expression Omnibus (GEO). GEO2R was employed to obtain differentially expressed circRNA between CC tissues and matched paracancerous tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were employed to detect circ_0000228, microRNA-337-3p (miR-337-3p) and transforming growth factor, beta receptor I (TGFBR1) expression levels in the CC tissues and cells. Following gain-of-function and loss-of-function models establishment, CCK-8 and BrdU tests were conducted to examine cell proliferation. Transwell experiment was executed to examine CC cells migration and invasion. A lung metastasis model was utilized to determine the ability of circ_0000228 on the lung metastasis. Bioinformatics analysis, dual-luciferase reporter experiment and RNA immunoprecipitation (RIP) assay were applied to verify the targeting relationship among miR-337-3p, circ_0000228, and TGFBR1. Results Circ_0000228 expression in the CC tissues and cells was up-modulated. Circ_0000228 overexpression markedly enhanced cell proliferation, migration, and invasion, while knocking down circ_0000228 remarkably repressed cell proliferation, migration, and invasion. MiR-337-3p could be adsorbed by circ_0000228. TGFBR1 was identified as a target gene of miR-337-3p that indirectly and positively modulated by circ_0000228 in the CC cells. Conclusion Circ_0000228 up-modulates TGFBR1 by targeting miR-337-3p to enhance CC cell proliferation, migration and invasion. Also, Circ_0000228 is a promising therapeutic target for the CC.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80360558","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Upregulation of hsa-miR-625-5p Inhibits Invasion of Acute Myeloid Leukemia Cancer Cells through ILK/AKT Pathway 上调hsa-miR-625-5p通过ILK/AKT通路抑制急性髓系白血病细胞的侵袭
Pub Date : 2022-02-01 DOI: 10.22074/cellj.2022.7658
Sahar Samieyan Dehkordi, S. Mousavi, Marzieh Ebrahimi, S. Alizadeh, Amir Abbas Hedayati Asl, Monireh Mohammad, Bahareh Aliabedi
Objective Acute myeloid leukemia (AML) is characterized by abnormalities of differentiation and growth of primary hematopoietic stem cells (HSCs) in the blood and bone marrow. In many studies, miR-625-5p has been shown to inhibit downstream pathways from affecting the metastasis and invasion of the integrin-linked kinase (ILK) signaling pathway. It has been proved that the expression of miR-625-5p decreases in AML cell lines. This study aimed to investigate the effect of miR-625-5p upregulation on the invasion of KG1 ell line in vitro. Materials and Methods In this experimental study, we investigated the impact of upregulation of miR-625-5p on invasion via the ILK/AKT pathway in the KG1 cell line. After transfection using the viral method, the cellular invasion was assessed by invasion assay and the levels of miR-625-5p genes and protein were evaluated by quantitative polymerase chain reaction (qPCR) and western blotting. Moreover, CXCR4 level was assessed by flow cytometry. Results The invasion significantly reduced in MiR-625-5p-transfected KG1 cells (P<0.01) that was concomitant with remarkably decreasing in the expression levels of ILK, NF-κB, and COX2 genes compare with the control group (P<0.01). Incontrast, MMP9, AP1, and AKT significantly increased (P<0.01, P<0.001 and P<0.01, respectively) and GSK3β did not change significantly in MiR-625-5p-transfected KG1 cells. The protein level of NF-κB decreased (P<0.01) and MMP9 increased, however it was not significant. Moreoever, the expression of CXCR4 was significantly lower (P<0.01) in comparison with the control group. Conclusion miR-625-5p leads to a reduction in cell invasion in the AML cell line through ILK pathway. Therefore, it could be a breakthrough in future AML-related research. However, further studies are needed to support this argument.
目的急性髓性白血病(Acute myeloid leukemia, AML)以造血干细胞(primary hematopoietic stem cells, hsc)在血液和骨髓中的分化和生长异常为特征。在许多研究中,miR-625-5p已被证明可以抑制下游通路影响整合素连接激酶(ILK)信号通路的转移和侵袭。已经证明,miR-625-5p在AML细胞系中表达降低。本研究旨在探讨miR-625-5p上调对体外KG1细胞系侵袭的影响。材料和方法在本实验研究中,我们通过ILK/AKT通路研究了miR-625-5p上调对KG1细胞系侵袭的影响。采用病毒法转染后,采用侵袭试验评估细胞侵袭,采用定量聚合酶链反应(qPCR)和western blotting检测miR-625-5p基因和蛋白水平。流式细胞术检测CXCR4水平。结果mir -625-5p转染的KG1细胞侵袭性明显降低(P<0.01), ILK、NF-κB、COX2基因表达水平明显低于对照组(P<0.01)。相比之下,转染mir -625-5p的KG1细胞中MMP9、AP1和AKT显著升高(分别为P<0.01、P<0.001和P<0.01), GSK3β无显著变化。NF-κB蛋白水平降低(P<0.01), MMP9蛋白水平升高,但差异不显著。与对照组相比,CXCR4的表达显著降低(P<0.01)。结论miR-625-5p通过ILK途径导致AML细胞系细胞侵袭减少。因此,这可能是未来aml相关研究的一个突破。然而,需要进一步的研究来支持这一论点。
{"title":"Upregulation of hsa-miR-625-5p Inhibits Invasion of Acute Myeloid Leukemia Cancer Cells through ILK/AKT Pathway","authors":"Sahar Samieyan Dehkordi, S. Mousavi, Marzieh Ebrahimi, S. Alizadeh, Amir Abbas Hedayati Asl, Monireh Mohammad, Bahareh Aliabedi","doi":"10.22074/cellj.2022.7658","DOIUrl":"https://doi.org/10.22074/cellj.2022.7658","url":null,"abstract":"Objective Acute myeloid leukemia (AML) is characterized by abnormalities of differentiation and growth of primary hematopoietic stem cells (HSCs) in the blood and bone marrow. In many studies, miR-625-5p has been shown to inhibit downstream pathways from affecting the metastasis and invasion of the integrin-linked kinase (ILK) signaling pathway. It has been proved that the expression of miR-625-5p decreases in AML cell lines. This study aimed to investigate the effect of miR-625-5p upregulation on the invasion of KG1 ell line in vitro. Materials and Methods In this experimental study, we investigated the impact of upregulation of miR-625-5p on invasion via the ILK/AKT pathway in the KG1 cell line. After transfection using the viral method, the cellular invasion was assessed by invasion assay and the levels of miR-625-5p genes and protein were evaluated by quantitative polymerase chain reaction (qPCR) and western blotting. Moreover, CXCR4 level was assessed by flow cytometry. Results The invasion significantly reduced in MiR-625-5p-transfected KG1 cells (P<0.01) that was concomitant with remarkably decreasing in the expression levels of ILK, NF-κB, and COX2 genes compare with the control group (P<0.01). Incontrast, MMP9, AP1, and AKT significantly increased (P<0.01, P<0.001 and P<0.01, respectively) and GSK3β did not change significantly in MiR-625-5p-transfected KG1 cells. The protein level of NF-κB decreased (P<0.01) and MMP9 increased, however it was not significant. Moreoever, the expression of CXCR4 was significantly lower (P<0.01) in comparison with the control group. Conclusion miR-625-5p leads to a reduction in cell invasion in the AML cell line through ILK pathway. Therefore, it could be a breakthrough in future AML-related research. However, further studies are needed to support this argument.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80289932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Safety and Efficacy of Allogeneic Adipose Tissue Mesenchymal Stromal Cells in Amyotrophic Lateral Sclerosis Patients, Single-Center, Prospective, Open-Label, Single-Arm Clinical Trial, Long-Term Follow-up 同种异体脂肪组织间充质基质细胞治疗肌萎缩性侧索硬化症患者的安全性和有效性,单中心、前瞻性、开放标签、单臂临床试验,长期随访
Pub Date : 2021-12-01 DOI: 10.22074/cellj.2021.7984
S. Nabavi, S. Karimi, L. Arab, L. Sanjari, S. Mardpour, V. Azimian, Neda Jarughi, A. Ghaheri, S. Hosseini, N. Aghdami, Massoud Vosough
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder with very limited treatment options. Stem cells have been raised as a new treatment modality for these patients. We have designed a single-center, prospective, open-label, and single arm clinical trial to assess the safety, feasibility, and rather efficacy of administrating allogeneic adipose-derived mesenchymal stromal cells (Ad-MSCs) in ALS patients. We enrolled 17 patients with confirmed ALS diagnosis with ALS Functional Rating Scale-Revised (ALSFRS-R) (<24) and predicted forced vital capacity (FVC) (<40)%. Allogeneic Ad-MSCs were transplanted intravenously for all patients. Follow-ups were done at 24 hours, 2, 4, 6, and 12 months after cell infusion by checking adverse events, laboratory tests, and clinically by ALSFRS-R and FVC. Patients were also followed five years later and ALSFRS-R score was recorded in the survived individuals. There was no report of severe adverse events related to cell infusion. Two patients experienced dyspnea and chest pain 36 and 65 days after cell infusion due to pulmonary emboli. The progressive decrease in ALSFRS-R and FVC levels was recorded and three patients died in the first year. During five years follow up, despite a notable decrease in functional scores, 5 patients survived. Intravenous (IV) infusion of allogeneic Ad-MSCs in ALS patients is safe and feasible. The survival rate of the patients is more than IV autologous MSCs (Registration number: IRCT20080728001031N26).
肌萎缩性侧索硬化症(ALS)是一种致命的神经退行性疾病,治疗方案非常有限。干细胞已被提出作为一种新的治疗方式。我们设计了一项单中心、前瞻性、开放标签、单臂临床试验,以评估同种异体脂肪源性间充质间质细胞(Ad-MSCs)治疗ALS患者的安全性、可行性和有效性。我们用ALS功能评定量表-修订版(ALSFRS-R)(<24)和预测用力肺活量(FVC)(<40)%纳入17例确诊为ALS的患者。所有患者均静脉移植同种异体Ad-MSCs。分别于细胞输注后24小时、2、4、6、12个月进行随访,检查不良事件、实验室检查和临床ALSFRS-R和FVC。5年后对患者进行随访,记录存活个体的ALSFRS-R评分。没有与细胞输注相关的严重不良事件的报道。2例患者因肺栓塞在细胞输注后36天和65天出现呼吸困难和胸痛。ALSFRS-R和FVC水平逐渐下降,3例患者在第一年死亡。在5年的随访中,尽管功能评分明显下降,但仍有5例患者存活。静脉(IV)输注同种异体Ad-MSCs在ALS患者中是安全可行的。患者生存率高于IV自体MSCs(注册号:IRCT20080728001031N26)。
{"title":"Safety and Efficacy of Allogeneic Adipose Tissue Mesenchymal Stromal Cells in Amyotrophic Lateral Sclerosis Patients, Single-Center, Prospective, Open-Label, Single-Arm Clinical Trial, Long-Term Follow-up","authors":"S. Nabavi, S. Karimi, L. Arab, L. Sanjari, S. Mardpour, V. Azimian, Neda Jarughi, A. Ghaheri, S. Hosseini, N. Aghdami, Massoud Vosough","doi":"10.22074/cellj.2021.7984","DOIUrl":"https://doi.org/10.22074/cellj.2021.7984","url":null,"abstract":"Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder with very limited treatment options. Stem cells have been raised as a new treatment modality for these patients. We have designed a single-center, prospective, open-label, and single arm clinical trial to assess the safety, feasibility, and rather efficacy of administrating allogeneic adipose-derived mesenchymal stromal cells (Ad-MSCs) in ALS patients. We enrolled 17 patients with confirmed ALS diagnosis with ALS Functional Rating Scale-Revised (ALSFRS-R) (<24) and predicted forced vital capacity (FVC) (<40)%. Allogeneic Ad-MSCs were transplanted intravenously for all patients. Follow-ups were done at 24 hours, 2, 4, 6, and 12 months after cell infusion by checking adverse events, laboratory tests, and clinically by ALSFRS-R and FVC. Patients were also followed five years later and ALSFRS-R score was recorded in the survived individuals. There was no report of severe adverse events related to cell infusion. Two patients experienced dyspnea and chest pain 36 and 65 days after cell infusion due to pulmonary emboli. The progressive decrease in ALSFRS-R and FVC levels was recorded and three patients died in the first year. During five years follow up, despite a notable decrease in functional scores, 5 patients survived. Intravenous (IV) infusion of allogeneic Ad-MSCs in ALS patients is safe and feasible. The survival rate of the patients is more than IV autologous MSCs (Registration number: IRCT20080728001031N26).","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78292908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 2
Restoration of MiR-34a Expression by 5-Azacytidine Augments Alimta -Induced Cell Death in Non-Small Lung Cancer Cells by Downregulation of HMG B1, A2 and Bcl-2 Pathway 5-氮胞苷恢复MiR-34a表达通过下调HMG B1、A2和Bcl-2通路增加Alimta诱导的非小肺癌细胞死亡
Pub Date : 2021-11-01 DOI: 10.22074/cellj.2021.7332.
Somayeh Amiri, Azra Rabbani-Chadegani, J. Davoodi, Hoda Gol Fakhrabadi
Objective Alimta (Pemetrexed) as an antifolate drug has been approved for the treatment of lung cancer. The aim of the present study was to investigate the combination effect of 5-Azacytidine (5-aza) and Alimta on the miR-34a and its target genes expression and induction of apoptotic cell death in non-small lung cancer A549 cells. Materials and Methods In this experimental study, lung cancer A549 cells were treated with various concentrations of Alimta alone and combined with 5-Aza. Then, viability was assessed by trypan blue and MTT assays. mRNA expressions were performed by real time-polymerase chain reaction (PCR) and western blot. Flow cytometry used to detect apoptotic/ necrotic cells and cell cycle arrest. Results Alimta alone reduced viability of the cells in a dose dependent manner with the half-maximal inhibitory concentration (IC50) value of 12 µM. Pretreatment of the cells with 5-aza (5 µM) induced a synergistic cytotoxic effect with IC50of 3 µM. Sequential exposure of the cells to 5-aza and Alimta enhanced miR-34a expression and significantly downregulated HMGB1, HMGA2 and BCL-2 expressions. Also, it was associated with reduction of nuclear HMGB1 and HMGA2 content. Caspase-3 activation, HMGB1 release into extracellular space and staining of the cells with annexine V/PI suggested that 5-aza reduced late apoptotic/necrotic cell death induced by Alimta. In addition, combination of 5-aza and Alimta arrested the cells at S and sub-G1 phases and inhibited colony formation. Conclusion 5-aza synergistically enhances Alimta induced apoptotic cell death through HMG proteins regulation, MIR34A gene expression and intrinsic apoptosis mechanism, providing a promising combination therapy in clinical lung cancer therapy.
目的阿利姆塔(培美曲塞)作为抗叶酸药物已被批准用于肺癌的治疗。本研究旨在探讨5-氮杂胞苷(5-aza)与Alimta联合作用对非小细胞肺癌A549细胞中miR-34a及其靶基因表达及诱导凋亡细胞死亡的影响。材料与方法本实验采用不同浓度的Alimta单独或联合5-Aza治疗肺癌A549细胞。然后用台盼蓝法和MTT法测定细胞活力。实时聚合酶链反应(real - time polymerase chain reaction, PCR)和western blot检测mRNA的表达。流式细胞术用于检测凋亡/坏死细胞和细胞周期阻滞。结果Alimta单用能明显降低细胞活力,IC50值为12µM,呈剂量依赖性。5-aza(5µM)预处理细胞具有协同细胞毒作用,ic50为3µM。细胞连续暴露于5-aza和Alimta后,miR-34a表达增强,HMGB1、HMGA2和BCL-2表达显著下调。与核HMGB1和HMGA2含量的减少有关。Caspase-3激活、HMGB1释放到细胞外空间以及annexine V/PI染色表明,5-aza减少了Alimta诱导的晚期凋亡/坏死细胞死亡。此外,5-aza和Alimta的联合抑制了S期和亚g1期的细胞,并抑制了集落的形成。结论5-aza通过HMG蛋白调控、MIR34A基因表达及内在凋亡机制协同增强Alimta诱导的凋亡细胞死亡,为临床肺癌治疗提供了一种有前景的联合治疗方案。
{"title":"Restoration of MiR-34a Expression by 5-Azacytidine Augments Alimta -Induced Cell Death in Non-Small Lung Cancer Cells by Downregulation of HMG B1, A2 and Bcl-2 Pathway","authors":"Somayeh Amiri, Azra Rabbani-Chadegani, J. Davoodi, Hoda Gol Fakhrabadi","doi":"10.22074/cellj.2021.7332.","DOIUrl":"https://doi.org/10.22074/cellj.2021.7332.","url":null,"abstract":"Objective Alimta (Pemetrexed) as an antifolate drug has been approved for the treatment of lung cancer. The aim of the present study was to investigate the combination effect of 5-Azacytidine (5-aza) and Alimta on the miR-34a and its target genes expression and induction of apoptotic cell death in non-small lung cancer A549 cells. Materials and Methods In this experimental study, lung cancer A549 cells were treated with various concentrations of Alimta alone and combined with 5-Aza. Then, viability was assessed by trypan blue and MTT assays. mRNA expressions were performed by real time-polymerase chain reaction (PCR) and western blot. Flow cytometry used to detect apoptotic/ necrotic cells and cell cycle arrest. Results Alimta alone reduced viability of the cells in a dose dependent manner with the half-maximal inhibitory concentration (IC50) value of 12 µM. Pretreatment of the cells with 5-aza (5 µM) induced a synergistic cytotoxic effect with IC50of 3 µM. Sequential exposure of the cells to 5-aza and Alimta enhanced miR-34a expression and significantly downregulated HMGB1, HMGA2 and BCL-2 expressions. Also, it was associated with reduction of nuclear HMGB1 and HMGA2 content. Caspase-3 activation, HMGB1 release into extracellular space and staining of the cells with annexine V/PI suggested that 5-aza reduced late apoptotic/necrotic cell death induced by Alimta. In addition, combination of 5-aza and Alimta arrested the cells at S and sub-G1 phases and inhibited colony formation. Conclusion 5-aza synergistically enhances Alimta induced apoptotic cell death through HMG proteins regulation, MIR34A gene expression and intrinsic apoptosis mechanism, providing a promising combination therapy in clinical lung cancer therapy.","PeriodicalId":9692,"journal":{"name":"Cell Journal (Yakhteh)","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78024391","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Decellularized Lung Extracellular Matrix Scaffold Promotes Human Embryonic Stem Cell Differentiation towards Alveolar Progenitors 脱细胞肺细胞外基质支架促进人胚胎干细胞向肺泡祖细胞分化
Pub Date : 2021-06-21 DOI: 10.21203/rs.3.rs-618083/v1
Afshin Noori, M. M. Mokhber Dezfouli, Sareh Rajabi, F. Ganji, Z. Ghezelayagh, E. El Agha, H. Baharvand, S. Sadeghian Chaleshtori, Yaser Tahamtani
Objective: Efficient production of functional and mature alveolar epithelial is a major challenge for developing any cell replacement therapy for lung degenerative diseases. The extracellular matrix (ECM) pro-vides a dynamic environment and mediates cellular responses during development and maintenance of tissue functions. The decellularized ECM (dECM) which retains its native-like structure and bio-chemical composition can provide the induction of embryonic stem cell (ESC) differentiation toward the tissue-specific lineages during in vitro culture. Therefore, the aim of this study was to evaluate the effect of sheep lung dECM-derived scaffold on differentiation and further maturation of ESC-derived lung progenitor cells. Materials and Methods: This study was an experimental study. In the first step, a sheep lung was decellularized to achieve dECM scaffolds and hydrogels. Afterwards, the obtained dECM scaffold was evaluated for collagen and glycosaminoglycan contents, DNA quantification, and its ultrastructure. Next, the three experimental groups: i. Sheep lung dECM-derived scaffold, ii. Sheep lung dECM-derived hydrogel, and iii. Fibronectin-coated plates were compared in their abilities to induce further differentiation of human embryonic stem cells (hESCs)-derived definitive endoderm (DE) into lung progenitor cells. The comparison was evaluated by immuno-staining and real-time polymerase chain reaction (PCR) assessments. Results: We found that the dECM-derived scaffold preserved its composition and native porous structures while lacking nuclei and intact cells. All experimental groups displayed lung progenitor cell differen-tiation as revealed by the RNA and protein expression of NKX2.1, P63 and CK5. DE cells differenti-ated on dECM-derived scaffold and dECMderived hydrogel showed significant upregulation of SOX9 gene expression, a marker of the distal airway epithelium. DE cells differentiated on the dECM-derived scaffold compared to the two other groups, showed enhanced expression of SFTPC (type 2 alveolar epithelial [AT2] cell marker), FOXJ1 (ciliated cell marker), and MUC5A (secretory cell marker) genes. Conclusion: Overall, our results suggest that dECM-derived scaffold improves the differentiation of DE cells towards lung alveolar progenitor cells in comparison with dECM-derived hydrogel and fibronectin-coated plates.
目的:高效生产功能成熟的肺泡上皮细胞是开发肺退行性疾病细胞替代疗法的主要挑战。细胞外基质(ECM)提供了一个动态环境,并在组织功能的发育和维持过程中介导细胞反应。在体外培养过程中,脱细胞ECM (dECM)保留了其原生结构和生化成分,可诱导胚胎干细胞(ESC)向组织特异性谱系分化。因此,本研究的目的是评估羊肺decm衍生支架对esc衍生肺祖细胞分化和进一步成熟的影响。材料与方法:本研究为实验研究。在第一步中,羊肺被去细胞化以获得dECM支架和水凝胶。然后,对所获得的dECM支架进行胶原蛋白和糖胺聚糖含量、DNA定量和超微结构评价。接下来,三个实验组:1 .羊肺decm衍生支架;羊肺decm衍生水凝胶;比较了纤维连接蛋白包被板诱导人胚胎干细胞(hESCs)来源的最终内胚层(DE)进一步分化为肺祖细胞的能力。通过免疫染色和实时聚合酶链反应(PCR)评估比较。结果:我们发现decm衍生的支架保留了其组成和天然多孔结构,但缺乏细胞核和完整的细胞。NKX2.1、P63和CK5的RNA和蛋白表达表明,各实验组均出现肺祖细胞分化。在decm来源的支架和decm来源的水凝胶上分化的DE细胞显示SOX9基因表达显著上调,SOX9基因是远端气道上皮的标志。与其他两组相比,在decm衍生支架上分化的DE细胞表现出SFTPC(2型肺泡上皮[AT2]细胞标志物)、FOXJ1(纤毛细胞标志物)和MUC5A(分泌细胞标志物)基因的表达增强。结论:总的来说,我们的研究结果表明,与decm来源的水凝胶和纤维连接蛋白包被板相比,decm来源的支架可以促进DE细胞向肺泡祖细胞的分化。
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引用次数: 0
Hsa-miR-587 Regulates TGFβ/SMAD Signaling and Promotes Cell Cycle Progression. Hsa-miR-587调控TGFβ/SMAD信号传导并促进细胞周期进展
Pub Date : 2020-07-01 Epub Date: 2019-10-14 DOI: 10.22074/cellj.2020.6483
Mahnaz Jahangirimoez, Abdallah Medlej, Mahmoud Tavallaie, Bahram Mohammad Soltani

Objective: Transforming growth factor beta/single mothers against decapentaplegic (TGFβ/SMAD) signaling pathway plays important roles in various biological processes. It acts as a tumor suppressor during the early stages of cancer progression. Discovering the regulators of this pathway provides important options for therapeutic strategies. Here, we searched for candidate microRNAs (miRNAs) that potentially target the critical components of the TGFβ signaling pathway.

Materials and methods: In the current experimental study, we first predicted miRNAs that target TGFβ components using a bioinformatics software. After that, quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expression of miR-587, TGFBR2, SMAD4, p21, CCND1 and c-MYC genes in transfected HEK293T and HCT116 cells. Dual Luciferase assay was performed to analyze the interactions between miRNAs and the target genes. Propidium iodide flow cytometry was used to determine cell cycle progression in HEK293T and HCT116 cells under hsa-miR-587 (miR-587) overexpression circumstances.

Results: Multiple miRNA responsive elements (MREs) were predicted for miR-587 within the 3'UTRs of the TGFBR2 and SMAD4 genes. Overexpression of miR-587 in HEK293T and HCT116 cells resulted in downregulation of TGFBR2 and SMAD4 genes. In addition, a downstream target gene of TGFβ/SMAD signaling, P21, was significantly downregulated in the HCT116 cells overexpressing miR-587. Dual luciferase assay analysis provided evidence that there is a direct interaction between miR-587 and the 3'UTR sequences of TGFBR2 and SMAD4 genes. Moreover, miR-587 overexpression in HEK293T and HCT116 cells resulted in reducing the SubG1 cell populations in both cell lines, as detected by flow cytometry.

Conclusion: Altogether, our data revealed an important role for miR-587 in regulating TGFβ/SMAD signaling and promoting cell cycle progression. These characteristics suggest that miR-587 is an important candidate for cancer therapy research.

目的:转化生长因子β/单体抗瘫痪母亲(TGFβ/SMAD)信号通路在各种生物过程中发挥着重要作用。在癌症进展的早期阶段,它是一种肿瘤抑制因子。发现该通路的调控因子为治疗策略提供了重要选择。在此,我们寻找了可能靶向 TGFβ 信号通路关键组分的候选 microRNA(miRNA):在目前的实验研究中,我们首先使用生物信息学软件预测了靶向 TGFβ 成分的 miRNAs。然后,使用实时定量聚合酶链反应(RT-qPCR)检测转染 HEK293T 和 HCT116 细胞中 miR-587、TGFBR2、SMAD4、p21、CCND1 和 c-MYC 基因的表达。采用双荧光素酶检测法分析 miRNA 与靶基因之间的相互作用。碘化丙啶流式细胞术用于确定在 hsa-miR-587 (miR-587)过表达情况下 HEK293T 和 HCT116 细胞的细胞周期进展:结果:在TGFBR2和SMAD4基因的3'UTR中预测了miR-587的多个miRNA应答元件(MREs)。在 HEK293T 和 HCT116 细胞中过表达 miR-587 会导致 TGFBR2 和 SMAD4 基因下调。此外,在过表达 miR-587 的 HCT116 细胞中,TGFβ/SMAD 信号转导的下游靶基因 P21 被显著下调。双荧光素酶分析证明,miR-587 与 TGFBR2 和 SMAD4 基因的 3'UTR 序列之间存在直接相互作用。此外,通过流式细胞术检测,在 HEK293T 和 HCT116 细胞中过表达 miR-587 能减少这两种细胞系中 SubG1 细胞的数量:总之,我们的数据揭示了 miR-587 在调节 TGFβ/SMAD 信号传导和促进细胞周期进展中的重要作用。这些特点表明,miR-587 是癌症治疗研究的重要候选对象。
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引用次数: 0
The Dilemma of TP53 Codon 72 Polymorphism (rs1042522) and Breast Cancer Risk: A Case-Control Study and Meta-Analysis in The Iranian Population TP53密码子72多态性(rs1042522)与乳腺癌风险的两难境地:伊朗人群的病例对照研究和荟萃分析
Pub Date : 2019-10-14 DOI: 10.22074/cellj.2020.6458
F. Afzaljavan, Negin Chaeichi Tehrani, M. Rivandi, Saeed Zarif Ghasemian, Elham Vahednia, Reza Khayami, Mohammadsajad Abavisani, Alireza Pasdar
Objective Mutations of TP53 as a tumor suppressor gene are frequently observed in different types of cancer. A codon 72 polymorphism located on exon 4 with two alleles encoding either Proline (CCC) or Arginine (CGC) has been indicated as a common variation in association with cancers. Controversial results have been reported regarding the association of allelic polymorphism of codon 72 of TP53 gene and breast cancer risk in Iranian patients. Therefore, a case-control study was designed. A meta-analysis was also carried out to provide evidence of association between this variation and breast cancer in Iran, based on all available published data. Materials and Methods In this case-control study, blood sample of 622 participants, including 308 breast cancer cases and 314 controls were collected. Genotyping for rs1042522 was conducted by Allele Specific polymerase chain reaction (AS-PCR). In order to set a meta-analysis study, PubMed, Scopus and ISI Web of Knowledge and Persian databases were searched to explore relevant studies, published up to September 2018, containing information on TP53 polymorphism and the risk of breast cancer in Iran. Statistical analysis was performed using SPSS 16.0 and MetaGenyo. Results All retrieved available data as well as the results of our current study were consisted of 1965 breast cancer cases and 1999 healthy controls. No significant difference was observed in allele frequencies between groups (P=0.90) in our study. The cumulative results did not also show any association between rs1042522 and breast cancer risk on the dominant (P=0.61) and recessive (P=0.89) models. Conclusion These findings cannot support contribution of rs1042522 polymorphism to breast cancer risk in an Iranian population. Future larger studies may help confirm this finding with a greater power. .
目的TP53作为肿瘤抑制基因,在不同类型的肿瘤中经常发生突变。位于外显子4上的密码子72多态性与两个编码脯氨酸(CCC)或精氨酸(CGC)的等位基因已被证明是与癌症相关的常见变异。关于TP53基因密码子72等位基因多态性与伊朗患者乳腺癌风险的关联,有争议的结果被报道。因此,设计了病例对照研究。基于所有可获得的已发表数据,还进行了一项荟萃分析,以提供这种变异与伊朗乳腺癌之间关联的证据。材料与方法在本病例对照研究中,收集了622名参与者的血液样本,其中包括308例乳腺癌患者和314例对照组。采用等位基因特异性聚合酶链反应(AS-PCR)对rs1042522进行基因分型。为了建立一项荟萃分析研究,检索了PubMed、Scopus、ISI Web of Knowledge和波斯语数据库,以探索截至2018年9月发表的相关研究,其中包含伊朗TP53多态性和乳腺癌风险的信息。采用SPSS 16.0和MetaGenyo进行统计学分析。结果所有检索到的数据以及本研究的结果包括1965例乳腺癌病例和1999例健康对照。本研究各组间等位基因频率差异无统计学意义(P=0.90)。累积结果也没有显示rs1042522与显性模型(P=0.61)和隐性模型(P=0.89)之间的任何关联。结论这些发现不能支持rs1042522多态性对伊朗人群乳腺癌风险的贡献。未来更大规模的研究可能有助于更有力地证实这一发现。
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引用次数: 5
期刊
Cell Journal (Yakhteh)
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