We previously showed that the benzoylphenylurea derivative BPU17 inhibits epithelial-mesenchymal transition and acts as an antifibrotic agent. This compound acts as a prohibitin (PHB) inhibitor by directly binding to PHB1. This binding disrupts the interaction between PHB1 and PHB2, leading to mild mitochondrial dysfunction. Here, we investigated the effect of BPU17 on angiogenesis using primary cultures of human vascular and microvascular endothelial cells, as well as a mouse model of choroidal neovascularization (CNV). A series of studies has shown that BPU17 inhibits angiogenesis both in vitro and in vivo. The molecular mechanism is that BPU17 inhibits serum response factor (SRF)/CArG box-mediated transcription by repressing the expression of SRF and its cofactor myocardin-related transcription factors (MRTF-A and -B [MRTF]). This defect causes the downregulation of adaptor and cell adhesion molecules such as vinculin and integrins, leading to the inhibition of angiogenesis. This inhibitory effect is closely associated with mild mitochondrial dysfunction, and siRNA-mediated knockdown of PHB1 similarly inhibits angiogenesis. Given that age-related inflammatory responses and subsequent choroidal neovascularization (CNV) contribute to the development of neovascular age-related macular degeneration (nAMD), this novel PHB inhibitor holds promise as a treatment for nAMD through its dual inhibitory effects on angiogenesis and fibrosis.
{"title":"A novel prohibitin inhibitor acts as a dual inhibitor of angiogenesis and fibrosis","authors":"Ken'ichiro Hayashi , Masaaki Kobayashi , Kotaro Mori , Yoshiaki Nakagawa , Bunta Watanabe , Takahiro Masuya , Atsushige Ashimori , Fumiaki Higashijima , Takuya Yoshimoto , Tsuyoshi Morita , Toshiyuki Murai , Saki Kirihara-Kojima , Kazuhiro Kimura","doi":"10.1016/j.bbrc.2026.153386","DOIUrl":"10.1016/j.bbrc.2026.153386","url":null,"abstract":"<div><div>We previously showed that the benzoylphenylurea derivative BPU17 inhibits epithelial-mesenchymal transition and acts as an antifibrotic agent. This compound acts as a prohibitin (PHB) inhibitor by directly binding to PHB1. This binding disrupts the interaction between PHB1 and PHB2, leading to mild mitochondrial dysfunction. Here, we investigated the effect of BPU17 on angiogenesis using primary cultures of human vascular and microvascular endothelial cells, as well as a mouse model of choroidal neovascularization (CNV). A series of studies has shown that BPU17 inhibits angiogenesis both in vitro and in vivo. The molecular mechanism is that BPU17 inhibits serum response factor (SRF)/CArG box-mediated transcription by repressing the expression of SRF and its cofactor myocardin-related transcription factors (MRTF-A and -B [MRTF]). This defect causes the downregulation of adaptor and cell adhesion molecules such as vinculin and integrins, leading to the inhibition of angiogenesis. This inhibitory effect is closely associated with mild mitochondrial dysfunction, and siRNA-mediated knockdown of PHB1 similarly inhibits angiogenesis. Given that age-related inflammatory responses and subsequent choroidal neovascularization (CNV) contribute to the development of neovascular age-related macular degeneration (nAMD), this novel PHB inhibitor holds promise as a treatment for nAMD through its dual inhibitory effects on angiogenesis and fibrosis.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153386"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-30DOI: 10.1016/j.bbrc.2026.153359
Hyun Sook Hwang , Mo Jong Kim , Sung Eun Kim , Eun-Kyoung Choi , Ji-Won Park
Senescence marker protein-30 (SMP-30; regucalcin) plays a crucial role in intracellular calcium homeostasis. This study investigated hepatic SMP-30 expression during non-alcoholic fatty liver disease (NAFLD) progression and evaluated the therapeutic potential of laminarin (LAM), a brown algae-derived polysaccharide, using high-fat diet (HFD)-fed mice and palmitic acid (PA)-treated Huh7 cells. Mice fed an HFD for 20 weeks developed NAFLD, characterized by elevated ALT/AST levels, hepatic steatosis, and significantly reduced SMP-30 expression. However, LAM treatment administered via drinking water (1%) or intraperitoneal injection (50 mg/kg) significantly attenuated lipid accumulation and restored hepatic SMP-30 expression. LAM reversed PA-induced lipid accumulation and SMP-30 downregulation in Huh7 cells. Mechanistically, LAM modulated the expression of SMP-30 and antioxidant proteins associated with activation of AKT/GSK3β/NRF2 signaling pathway, thereby mitigating the adverse effects of PA-induced toxicity. In conclusion, hepatic SMP-30 expression decreases during NAFLD progression, and LAM treatment restores these levels while alleviating lipid accumulation. These findings suggest that LAM may represent a promising therapeutic agent for NAFLD by improving lipid metabolism and reducing oxidative stress through the regulation of SMP-30.
{"title":"Laminarin ameliorates hepatic steatosis and upregulates hepatic SMP-30 expression in high-fat-diet-induced NAFLD mice","authors":"Hyun Sook Hwang , Mo Jong Kim , Sung Eun Kim , Eun-Kyoung Choi , Ji-Won Park","doi":"10.1016/j.bbrc.2026.153359","DOIUrl":"10.1016/j.bbrc.2026.153359","url":null,"abstract":"<div><div>Senescence marker protein-30 (SMP-30; regucalcin) plays a crucial role in intracellular calcium homeostasis. This study investigated hepatic SMP-30 expression during non-alcoholic fatty liver disease (NAFLD) progression and evaluated the therapeutic potential of laminarin (LAM), a brown algae-derived polysaccharide, using high-fat diet (HFD)-fed mice and palmitic acid (PA)-treated Huh7 cells. Mice fed an HFD for 20 weeks developed NAFLD, characterized by elevated ALT/AST levels, hepatic steatosis, and significantly reduced SMP-30 expression. However, LAM treatment administered via drinking water (1%) or intraperitoneal injection (50 mg/kg) significantly attenuated lipid accumulation and restored hepatic SMP-30 expression. LAM reversed PA-induced lipid accumulation and SMP-30 downregulation in Huh7 cells. Mechanistically, LAM modulated the expression of SMP-30 and antioxidant proteins associated with activation of AKT/GSK3β/NRF2 signaling pathway, thereby mitigating the adverse effects of PA-induced toxicity. In conclusion, hepatic SMP-30 expression decreases during NAFLD progression, and LAM treatment restores these levels while alleviating lipid accumulation. These findings suggest that LAM may represent a promising therapeutic agent for NAFLD by improving lipid metabolism and reducing oxidative stress through the regulation of SMP-30.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153359"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117607","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-28DOI: 10.1016/j.bbrc.2026.153374
Yury B. Slonimskiy , Daria A. Lunegova , Andrey S. Nikolaev , Ilya O. Matyuta , Konstantin E. Klementiev , Vladimir O. Popov , Konstantin M. Boyko , Ivan Yu. Gushchin , Nikolai N. Sluchanko
The recently discovered family of microalgal water-soluble astaxanthin-binding proteins (AstaPs) functions in carotenoid sequestration and transfer. We present the crystal structure and characterization of AstaP-pink1, a homolog from Scenedesmus sp. We show that despite low sequence identity with the known AstaP-orange1, AstaP-pink1 also binds various xanthophylls, but in contrast to AstaP-orange1, induces a pronounced bathochromic shift and a near-UV spectral bump upon binding ketocarotenoids, which is reminiscent of photoactivated Orange Carotenoid Protein (OCP). Mutagenesis and domain-swap experiments indicate that its unique spectral tuning likely results from carotenoid isomerization, forced by the interplay between its N-terminal jaw and body subdomains connected by the hinge loop of different sequence and length. This study attempts to rationalize the spectral and functional diversification of AstaPs and highlights their potential as engineered modules for carotenoid delivery.
{"title":"Crystal structure and functional characterization of the microalgal carotenoprotein AstaP-pink1","authors":"Yury B. Slonimskiy , Daria A. Lunegova , Andrey S. Nikolaev , Ilya O. Matyuta , Konstantin E. Klementiev , Vladimir O. Popov , Konstantin M. Boyko , Ivan Yu. Gushchin , Nikolai N. Sluchanko","doi":"10.1016/j.bbrc.2026.153374","DOIUrl":"10.1016/j.bbrc.2026.153374","url":null,"abstract":"<div><div>The recently discovered family of microalgal water-soluble astaxanthin-binding proteins (AstaPs) functions in carotenoid sequestration and transfer. We present the crystal structure and characterization of AstaP-pink1, a homolog from <em>Scenedesmus</em> sp. We show that despite low sequence identity with the known AstaP-orange1, AstaP-pink1 also binds various xanthophylls, but in contrast to AstaP-orange1, induces a pronounced bathochromic shift and a near-UV spectral bump upon binding ketocarotenoids, which is reminiscent of photoactivated Orange Carotenoid Protein (OCP). Mutagenesis and domain-swap experiments indicate that its unique spectral tuning likely results from carotenoid isomerization, forced by the interplay between its N-terminal jaw and body subdomains connected by the hinge loop of different sequence and length. This study attempts to rationalize the spectral and functional diversification of AstaPs and highlights their potential as engineered modules for carotenoid delivery.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153374"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117460","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-29DOI: 10.1016/j.bbrc.2026.153377
Ariunbold Chuluun-Erdene , Takahiro Kuchimaru , Takayuki Isagawa , Tatsuyuki Sato , Hiroki Sugimoto , Kazutoshi Ono , Daigo Sawaki , Shigeru Sato , Masamichi Yamamoto , Norihiko Takeda
Cardiac fibroblasts (CFs) are the predominant non-myocyte cell type in the heart and play central roles in extracellular matrix remodeling and intercellular signaling during cardiac physiology and pathology. However, the bioenergetic basis underlying CF functions remains poorly understood, mainly due to the lack of tools for visualizing intracellular adenosine triphosphate (ATP) dynamics with high spatiotemporal resolution. Here, we established immortalized murine cardiac fibroblasts stably expressing the genetically encoded ATP indicator GO-ATeam2 based on Förster Resonance Energy Transfer (FRET). The resulting CF7/GO-ATeam2 cell line allows real-time and quantitative monitoring of cytosolic ATP levels in living cells. CF7/GO-ATeam2 cells exhibited robust proliferation and quick responses to change of cytosolic ATP level. We demonstrated dynamic cytosolic ATP imaging upon pharmacological perturbations of oxidative phosphorylation and glycolysis, as well as under growth factor stimulation. Our work provides the CF7/GO-ATeam2 platform, a versatile cellular resource for dissecting the metabolic regulation of cardiac fibroblasts, offering new opportunities to explore energy dynamics in cardiac physiology and disease.
{"title":"Establishment of immortalized murine cardiac fibroblasts for visualizing cytosolic ATP dynamics with a genetically encoded optical indicator","authors":"Ariunbold Chuluun-Erdene , Takahiro Kuchimaru , Takayuki Isagawa , Tatsuyuki Sato , Hiroki Sugimoto , Kazutoshi Ono , Daigo Sawaki , Shigeru Sato , Masamichi Yamamoto , Norihiko Takeda","doi":"10.1016/j.bbrc.2026.153377","DOIUrl":"10.1016/j.bbrc.2026.153377","url":null,"abstract":"<div><div>Cardiac fibroblasts (CFs) are the predominant non-myocyte cell type in the heart and play central roles in extracellular matrix remodeling and intercellular signaling during cardiac physiology and pathology. However, the bioenergetic basis underlying CF functions remains poorly understood, mainly due to the lack of tools for visualizing intracellular adenosine triphosphate (ATP) dynamics with high spatiotemporal resolution. Here, we established immortalized murine cardiac fibroblasts stably expressing the genetically encoded ATP indicator GO-ATeam2 based on Förster Resonance Energy Transfer (FRET). The resulting CF7/GO-ATeam2 cell line allows real-time and quantitative monitoring of cytosolic ATP levels in living cells. CF7/GO-ATeam2 cells exhibited robust proliferation and quick responses to change of cytosolic ATP level. We demonstrated dynamic cytosolic ATP imaging upon pharmacological perturbations of oxidative phosphorylation and glycolysis, as well as under growth factor stimulation. Our work provides the CF7/GO-ATeam2 platform, a versatile cellular resource for dissecting the metabolic regulation of cardiac fibroblasts, offering new opportunities to explore energy dynamics in cardiac physiology and disease.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153377"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-29DOI: 10.1016/j.bbrc.2026.153352
Reham Barghash , Piyawan Chailapakul , Amber R. Prebble , Del Leary , Takamitsu A. Kato
This study evaluates the radiobiological effectiveness comparing superficial X-rays to Cs-137 in V79 and U2OS cells cultured in T25 flasks using colony formation assays. Additional dosimetric care was given to maintaining equal absolute dose for all beam qualities, correcting for absorption and scattering in the flask, and quantifiable interpretation due to dose of the spatial differences in cell survival with significantly higher survival near the flask's edges. Ion chamber measurements quantified flask lid attenuation (6.9 %, 2.4 %, and 1.9 % for 50, 70, and 100 kVp), enabling dose correction for accurate survival analysis. CT-based Monte Carlo simulations revealed thicker flask walls and the meniscus effect created a 35 % dose reduction from flask center to edges, directly explaining the observed survival patterns. Monte Carlo simulation also predicted an increased secondary electron production at higher energies. These results emphasize the necessity of precise dosimetry in low-energy X-ray studies. Combined experimental and computational approaches enhanced reliability of radiobiological assessments. Survival curves yielded Relative Biological Effectiveness (RBE) values for 50 kVp X-ray of 1.45 (V79) and 1.64 (U2OS); 1.41 (V79) and 1.55 (U2OS) at 70 kVp; and 1.23 (V79) and 1.36 (U2OS) at 100 kVp, relative to Cs-137.
{"title":"Biological effectiveness of superficial X-ray in mammalian cells through precise dosimetry and Monte Carlo simulations","authors":"Reham Barghash , Piyawan Chailapakul , Amber R. Prebble , Del Leary , Takamitsu A. Kato","doi":"10.1016/j.bbrc.2026.153352","DOIUrl":"10.1016/j.bbrc.2026.153352","url":null,"abstract":"<div><div>This study evaluates the radiobiological effectiveness comparing superficial X-rays to Cs-137 in V79 and U2OS cells cultured in T25 flasks using colony formation assays. Additional dosimetric care was given to maintaining equal absolute dose for all beam qualities, correcting for absorption and scattering in the flask, and quantifiable interpretation due to dose of the spatial differences in cell survival with significantly higher survival near the flask's edges. Ion chamber measurements quantified flask lid attenuation (6.9 %, 2.4 %, and 1.9 % for 50, 70, and 100 kVp), enabling dose correction for accurate survival analysis. CT-based Monte Carlo simulations revealed thicker flask walls and the meniscus effect created a 35 % dose reduction from flask center to edges, directly explaining the observed survival patterns. Monte Carlo simulation also predicted an increased secondary electron production at higher energies. These results emphasize the necessity of precise dosimetry in low-energy X-ray studies. Combined experimental and computational approaches enhanced reliability of radiobiological assessments. Survival curves yielded Relative Biological Effectiveness (RBE) values for 50 kVp X-ray of 1.45 (V79) and 1.64 (U2OS); 1.41 (V79) and 1.55 (U2OS) at 70 kVp; and 1.23 (V79) and 1.36 (U2OS) at 100 kVp, relative to Cs-137.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153352"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-02-02DOI: 10.1016/j.bbrc.2026.153389
Gaoyue Jiang , Chunxia Li , Baihui Jia , Meng Lu , Yuqing Lei , Kefeng Lu , Tao cui , Huihui Li
Objectives
Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer. Due to its radiotherapy resistance, high metastasis rate and recurrence rate, the prognosis is poor. So far, effective therapeutic drugs for it remain elusive. Cetylpyridinium chloride (CPC) has initially demonstrated anti-tumor properties in various tumors. Therefore, the aim of this study is to explore the intervention effect of CPC on macrophages in the tumor microenvironment (TME) and its therapeutic effect on TNBC.
Methods
The effects of CPC on macrophage polarization and expression changes were evaluated by WB and RT-qPCR. Multiple TNBC cell lines and non-cancer cells were exposed to CPC under different conditions (concentration and duration), and cell survival and proliferation were assessed by CCK8 and colony formation. The effects of CPC on apoptosis, migration and invasion of TNBC cells were further evaluated. The mitochondrial status was assessed by Mito-Tracker Red CMXRos staining, mitochondrial protein expression detection and ATP content detection to explain the cause of apoptosis. The effect of CPC on TNBC growth was further confirmed in animal models.
Results
Our results demonstrated that CPC (2 μM) inhibited the M2 polarization of macrophages induced by interleukin-4 (IL-4) and tumor secretions, and reduced their characteristic secretions that promote tumor growth. Meanwhile, CPC significantly suppressed the proliferation of multiple TNBC cell lines, induced mitochondrial damage, significantly decreased cellular ATP levels, and ultimately led to tumor cell apoptosis. Moreover, TNBC cells are more sensitive to CPC than non-tumor cells. Additionally, CPC significantly inhibited the migration and invasion of TNBC cells. Finally, we confirmed the growth inhibitory effect of CPC on TNBC in vivo.
Conclusion
CPC has dual effects: it inhibits the M2 polarization of tumor-associated macrophages (TAMs) and directly suppresses the proliferation and invasion of TNBC cells, ultimately effectively inhibiting the growth of TNBC in vivo. Considering that CPC has been administered in humans as a safe drug-disinfectant for decades, our study here provides a molecular basis for the application of CPC as a potential option to clinical treatments of TNBC.
目的：三阴性乳腺癌（TNBC）是最具侵袭性的乳腺癌亚型。由于其耐放疗、转移率和复发率高，预后较差。到目前为止，有效的治疗药物仍然难以捉摸。十六烷基氯化吡啶（CPC）在多种肿瘤中已初步显示出抗肿瘤特性。因此，本研究旨在探讨CPC对肿瘤微环境（tumor microenvironment， TME）中巨噬细胞的干预作用及其对TNBC的治疗作用。方法：采用WB法和RT-qPCR法观察CPC对巨噬细胞极化和表达变化的影响。将多种TNBC细胞系和非癌细胞在不同条件（浓度和持续时间）下暴露于CPC，通过CCK8和集落形成评估细胞存活和增殖。进一步观察CPC对TNBC细胞凋亡、迁移和侵袭的影响。通过Mito-Tracker Red CMXRos染色、线粒体蛋白表达检测和ATP含量检测评估线粒体状态，解释细胞凋亡的原因。在动物模型中进一步证实了CPC对TNBC生长的影响。结果：CPC（2 μM）可抑制白细胞介素-4 （IL-4）和肿瘤分泌物诱导的巨噬细胞M2极化，减少巨噬细胞促进肿瘤生长的特征性分泌物。同时，CPC显著抑制多种TNBC细胞株的增殖，诱导线粒体损伤，显著降低细胞ATP水平，最终导致肿瘤细胞凋亡。此外，TNBC细胞比非肿瘤细胞对CPC更敏感。此外，CPC显著抑制TNBC细胞的迁移和侵袭。最后，我们在体内证实了CPC对TNBC的生长抑制作用。结论：CPC具有双重作用：一方面抑制肿瘤相关巨噬细胞（tumor-associated macrophages, tam）的M2极化，另一方面直接抑制TNBC细胞的增殖和侵袭，最终在体内有效抑制TNBC的生长。考虑到CPC作为一种安全的药物消毒剂已经在人体中使用了几十年，我们的研究为CPC作为TNBC临床治疗的潜在选择提供了分子基础。
{"title":"Cetylpyridinium chloride inhibits the polarization of tumor-associated macrophages and the proliferation of triple-negative breast cancer cells","authors":"Gaoyue Jiang , Chunxia Li , Baihui Jia , Meng Lu , Yuqing Lei , Kefeng Lu , Tao cui , Huihui Li","doi":"10.1016/j.bbrc.2026.153389","DOIUrl":"10.1016/j.bbrc.2026.153389","url":null,"abstract":"<div><h3>Objectives</h3><div>Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer. Due to its radiotherapy resistance, high metastasis rate and recurrence rate, the prognosis is poor. So far, effective therapeutic drugs for it remain elusive. Cetylpyridinium chloride (CPC) has initially demonstrated anti-tumor properties in various tumors. Therefore, the aim of this study is to explore the intervention effect of CPC on macrophages in the tumor microenvironment (TME) and its therapeutic effect on TNBC.</div></div><div><h3>Methods</h3><div>The effects of CPC on macrophage polarization and expression changes were evaluated by WB and RT-qPCR. Multiple TNBC cell lines and non-cancer cells were exposed to CPC under different conditions (concentration and duration), and cell survival and proliferation were assessed by CCK8 and colony formation. The effects of CPC on apoptosis, migration and invasion of TNBC cells were further evaluated. The mitochondrial status was assessed by Mito-Tracker Red CMXRos staining, mitochondrial protein expression detection and ATP content detection to explain the cause of apoptosis. The effect of CPC on TNBC growth was further confirmed in animal models.</div></div><div><h3>Results</h3><div>Our results demonstrated that CPC (2 μM) inhibited the M2 polarization of macrophages induced by interleukin-4 (IL-4) and tumor secretions, and reduced their characteristic secretions that promote tumor growth. Meanwhile, CPC significantly suppressed the proliferation of multiple TNBC cell lines, induced mitochondrial damage, significantly decreased cellular ATP levels, and ultimately led to tumor cell apoptosis. Moreover, TNBC cells are more sensitive to CPC than non-tumor cells. Additionally, CPC significantly inhibited the migration and invasion of TNBC cells. Finally, we confirmed the growth inhibitory effect of CPC on TNBC in vivo.</div></div><div><h3>Conclusion</h3><div>CPC has dual effects: it inhibits the M2 polarization of tumor-associated macrophages (TAMs) and directly suppresses the proliferation and invasion of TNBC cells, ultimately effectively inhibiting the growth of TNBC in vivo. Considering that CPC has been administered in humans as a safe drug-disinfectant for decades, our study here provides a molecular basis for the application of CPC as a potential option to clinical treatments of TNBC.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153389"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146137069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-25DOI: 10.1016/j.bbrc.2026.153345
Yi Lei , Chenlin Gao , Xin Zhao , Betty Yuen Kwan Law , Yong Xu
Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease, driven by hyperglycemia-induced mitochondrial apoptosis in renal tubular epithelial cells. Death-associated protein kinase 1 (DAPK1) is a key mediator of cell death, but its regulation in DKD remains unclear. Here, we investigated the mechanisms underlying DAPK1 upregulation and its role in mitochondrial apoptosis under high glucose (HG) conditions in HK-2 cells and db/db mouse models. In db/db mice, renal DAPK1 protein levels were elevated, while KLHL20 levels were reduced, correlating with glomerular and tubular injury. In HK-2 cells, HG (33 mM, 48 h) significantly increased DAPK1 mRNA and protein levels while prolonging its half-life. Mechanistically, HG transcriptionally suppressed KLHL20, an E3 ubiquitin ligase adaptor that targets DAPK1 for proteasomal degradation. Co-immunoprecipitation confirmed KLHL20-DAPK1 interaction and showed reduced DAPK1 ubiquitination under HG. Overexpression of KLHL20 restored DAPK1 ubiquitination and reduced its protein levels without affecting mRNA, confirming post-translational regulation. Functionally, DAPK1 knockdown attenuated HG-induced mitochondrial apoptosis. KLHL20 overexpression similarly protected against HG-induced apoptosis, but this effect was abrogated by DAPK1 co-overexpression, establishing DAPK1 as a critical downstream effector. These findings reveal a novel KLHL20-DAPK1 axis where HG stabilizes DAPK1 by downregulating KLHL20, promoting mitochondrial apoptosis in renal tubular cells. Targeting this pathway may offer therapeutic strategies for DKD.
糖尿病肾病（DKD）是终末期肾脏疾病的主要原因，由高血糖诱导的肾小管上皮细胞线粒体凋亡驱动。死亡相关蛋白激酶1 （DAPK1）是细胞死亡的关键介质，但其在DKD中的调控尚不清楚。在此，我们在HK-2 细胞和db/db小鼠模型中研究了高糖（HG）条件下DAPK1上调的机制及其在线粒体凋亡中的作用。在db/db小鼠中，肾脏DAPK1蛋白水平升高，而KLHL20水平降低，与肾小球和小管损伤相关。在HK-2 细胞中，HG（33 mM, 48 h）显著提高了DAPK1 mRNA和蛋白水平，延长了其半衰期。在机制上，HG转录抑制了KLHL20，这是一种E3泛素连接酶适配器，靶向DAPK1进行蛋白酶体降解。共免疫沉淀证实了KLHL20-DAPK1相互作用，并显示HG下DAPK1泛素化降低。过表达KLHL20恢复了DAPK1泛素化，降低了其蛋白水平，但不影响mRNA，证实了翻译后调控。功能上，DAPK1敲低可减弱hg诱导的线粒体凋亡。KLHL20过表达同样可以防止hg诱导的细胞凋亡，但这种作用被DAPK1共过表达所消除，这表明DAPK1是一个关键的下游效应物。这些发现揭示了一个新的KLHL20-DAPK1轴，HG通过下调KLHL20来稳定DAPK1，促进肾小管细胞线粒体凋亡。靶向这一途径可能为DKD提供治疗策略。
{"title":"KLHL20 alleviates high glucose-induced mitochondrial apoptosis in renal tubular cells by targeting DAPK1 for ubiquitination and degradation","authors":"Yi Lei , Chenlin Gao , Xin Zhao , Betty Yuen Kwan Law , Yong Xu","doi":"10.1016/j.bbrc.2026.153345","DOIUrl":"10.1016/j.bbrc.2026.153345","url":null,"abstract":"<div><div>Diabetic kidney disease (DKD) is a leading cause of end-stage renal disease, driven by hyperglycemia-induced mitochondrial apoptosis in renal tubular epithelial cells. Death-associated protein kinase 1 (DAPK1) is a key mediator of cell death, but its regulation in DKD remains unclear. Here, we investigated the mechanisms underlying DAPK1 upregulation and its role in mitochondrial apoptosis under high glucose (HG) conditions in HK-2 cells and <em>db/db</em> mouse models. In <em>db/db</em> mice, renal DAPK1 protein levels were elevated, while KLHL20 levels were reduced, correlating with glomerular and tubular injury. In HK-2 cells, HG (33 mM, 48 h) significantly increased DAPK1 mRNA and protein levels while prolonging its half-life. Mechanistically, HG transcriptionally suppressed KLHL20, an E3 ubiquitin ligase adaptor that targets DAPK1 for proteasomal degradation. Co-immunoprecipitation confirmed KLHL20-DAPK1 interaction and showed reduced DAPK1 ubiquitination under HG. Overexpression of <em>KLHL20</em> restored DAPK1 ubiquitination and reduced its protein levels without affecting mRNA, confirming post-translational regulation. Functionally, <em>DAPK1</em> knockdown attenuated HG-induced mitochondrial apoptosis. <em>KLHL20</em> overexpression similarly protected against HG-induced apoptosis, but this effect was abrogated by <em>DAPK1</em> co-overexpression, establishing DAPK1 as a critical downstream effector. These findings reveal a novel KLHL20-DAPK1 axis where HG stabilizes DAPK1 by downregulating KLHL20, promoting mitochondrial apoptosis in renal tubular cells. Targeting this pathway may offer therapeutic strategies for DKD.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153345"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146117575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-01-29DOI: 10.1016/j.bbrc.2026.153331
Sama Torkashvand , Sohrab Kazemi , Ali Akbar Moghadamnia
Chemotherapy is a significant treatment for cancer; however, it is frequently associated with deleterious side effects. Thymoquinone (TQ), a naturally occurring compound, has been reported to exhibit notable anti-cancer potential. The present study focuses on the in vitro effects of TQ-loaded nanoemulsion (TQ-NE) as an independent treatment and alongside the prevalent chemotherapeutic agents, 5-fluorouracil (5-FU) and capecitabine (CAP), on a colorectal cancer (HT-29) cell line. A TQ-NE formulation was synthesized and characterized, revealing an average particle size of 129.5 nm. The IC50 values for CAP, 5-FU, TQ, NE (carrier), and TQ-NE were 39.57, 61.01, 21.81, 218.4, and 15.12 μM, respectively. Cytotoxic effects were examined in both HT-29 cells and fibroblasts, revealing that TQ-NE enhanced the sensitivity of cancer cells to chemotherapeutic agents. When combined with 5-FU or CAP, TQ-NE produced a greater reduction in cell viability than TQ alone, indicating synergistic interaction. Flow cytometry analysis further revealed that TQ-NE induced distinct apoptotic responses compared to the control, whereas its combination with chemotherapy drugs showed a different pattern.
These findings suggest that TQ-NE can enhance the cytotoxic activities of 5-FU and CAP in vitro. Furthermore, using lower concentrations of TQ-NE could potentiate the inhibitory effects of 5-FU and CAP, presenting a promising strategy for improving colorectal cancer (CRC) treatment outcomes while potentially reducing the adverse effects related to higher doses of conventional chemotherapy.
{"title":"Nanoemulsion in cancer therapy: Enhancing the therapeutic efficacy of capecitabine and 5-fluorouracil in colorectal cancer with thymoquinone nanoemulsion","authors":"Sama Torkashvand , Sohrab Kazemi , Ali Akbar Moghadamnia","doi":"10.1016/j.bbrc.2026.153331","DOIUrl":"10.1016/j.bbrc.2026.153331","url":null,"abstract":"<div><div>Chemotherapy is a significant treatment for cancer; however, it is frequently associated with deleterious side effects. Thymoquinone (TQ), a naturally occurring compound, has been reported to exhibit notable anti-cancer potential. The present study focuses on the <em>in vitro</em> effects of TQ-loaded nanoemulsion (TQ-NE) as an independent treatment and alongside the prevalent chemotherapeutic agents, 5-fluorouracil (5-FU) and capecitabine (CAP), on a colorectal cancer (HT-29) cell line. A TQ-NE formulation was synthesized and characterized, revealing an average particle size of 129.5 nm. The IC<sub>50</sub> values for CAP, 5-FU, TQ, NE (carrier), and TQ-NE were 39.57, 61.01, 21.81, 218.4, and 15.12 μM, respectively. Cytotoxic effects were examined in both HT-29 cells and fibroblasts, revealing that TQ-NE enhanced the sensitivity of cancer cells to chemotherapeutic agents. When combined with 5-FU or CAP, TQ-NE produced a greater reduction in cell viability than TQ alone, indicating synergistic interaction. Flow cytometry analysis further revealed that TQ-NE induced distinct apoptotic responses compared to the control, whereas its combination with chemotherapy drugs showed a different pattern.</div><div>These findings suggest that TQ-NE can enhance the cytotoxic activities of 5-FU and CAP <em>in vitro</em>. Furthermore, using lower concentrations of TQ-NE could potentiate the inhibitory effects of 5-FU and CAP, presenting a promising strategy for improving colorectal cancer (CRC) treatment outcomes while potentially reducing the adverse effects related to higher doses of conventional chemotherapy.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153331"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146123538","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-02-04DOI: 10.1016/j.bbrc.2026.153406
Shoukang Du , Yuhan Wang , Ting Gang Chew
Cells maintain proteostasis by sequestering misfolded proteins into deposition sites. Aggregation-prone endoplasmic reticulum (ER) proteins form membrane-bound nuclear compartments that are cleared during cell division, yet the mechanisms underlying their spatial organization remain unclear. Here, using transcriptomic and proteomic analyses, we identified the ER-localized Hsp70 chaperone BiP as a key player. Genetic depletion or chemical inhibition of BiP prevented nuclear aggregate formation, while manipulating BiP regulators perturbed the aggregate formation. BiP-driven aggregation precedes the inner nuclear membrane synthesis that encapsulated the aggregates. Under proteostatic stress, nuclear aggregates localized adjacent to ER-derived aggregates. Our findings demonstrate that BiP is essential for organizing ER-derived aggregates in the nucleus, which further regulate nuclear proteostasis through spatial interactions with nuclear aggregates.
{"title":"Spatial organization of ER-derived protein aggregates in the nucleus requires the Hsp70-family member BiP","authors":"Shoukang Du , Yuhan Wang , Ting Gang Chew","doi":"10.1016/j.bbrc.2026.153406","DOIUrl":"10.1016/j.bbrc.2026.153406","url":null,"abstract":"<div><div>Cells maintain proteostasis by sequestering misfolded proteins into deposition sites. Aggregation-prone endoplasmic reticulum (ER) proteins form membrane-bound nuclear compartments that are cleared during cell division, yet the mechanisms underlying their spatial organization remain unclear. Here, using transcriptomic and proteomic analyses, we identified the ER-localized Hsp70 chaperone BiP as a key player. Genetic depletion or chemical inhibition of BiP prevented nuclear aggregate formation, while manipulating BiP regulators perturbed the aggregate formation. BiP-driven aggregation precedes the inner nuclear membrane synthesis that encapsulated the aggregates. Under proteostatic stress, nuclear aggregates localized adjacent to ER-derived aggregates. Our findings demonstrate that BiP is essential for organizing ER-derived aggregates in the nucleus, which further regulate nuclear proteostasis through spatial interactions with nuclear aggregates.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153406"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-03-19Epub Date: 2026-02-02DOI: 10.1016/j.bbrc.2026.153392
Jinglian Mo , Ziming Liu , Jialin Fu , Qichao Ge , Yiying Qu , Nan Ge , Yang Liu
Background
Proline supplementation promotes proliferation and invasion in cancers under acute nutrient deprivation. However, its molecular basis in hepatocellular carcinoma is not fully understood yet. This study investigates how proline metabolism promotes hepatocellular carcinoma (HCC) cell proliferation and survival during nutrient starvation.
Methods
Proline metabolism-related protein expression in HCC versus paracarcinoma tissues was analyzed using The Cancer Genome Atlas database. HCC cell viability was quantified via Cell Counting Kit 8 assays, and colony formation capacity was evaluated. Intracellular ROS levels were measured by flow cytometry. Autophagic flux was assessed by GFP/mCherry fluorescence ratio and autophagy-related proteins by immunoblotting. Lipid droplet deposition was visualized using Hoechst 33342 and BODIPY 493/503 staining.
Results
Proline metabolism-related proteins were significantly upregulated in HCC tissues compared to paracarcinoma controls. Under acute nutrient stress, exogenous proline reduced cellular ROS levels, attenuated lipid droplet accumulation, and suppressed excessive autophagy in HCC cells. Proline rescued nutrient deprivation-induced tumor growth inhibition, which could be reversed by H2O2 and rapamycin.
Conclusion
Proline metabolism sustains HCC cell survival during nutrient restriction by reducing ROS accumulation, thereby inhibiting lipid droplet formation and autophagy.
背景：补充脯氨酸促进急性营养剥夺下癌症的增殖和侵袭。然而，其在肝细胞癌中的分子基础尚不完全清楚。本研究旨在探讨脯氨酸代谢如何促进肝细胞癌（HCC）细胞在营养缺乏状态下的增殖和存活。方法：使用The Cancer Genome Atlas数据库分析脯氨酸代谢相关蛋白在HCC和癌旁组织中的表达。通过细胞计数试剂盒8测定HCC细胞活力，并评估集落形成能力。流式细胞术检测细胞内ROS水平。采用GFP/mCherry荧光比测定自噬通量，免疫印迹法测定自噬相关蛋白。采用Hoechst 33342和BODIPY 493/503染色观察脂滴沉积。结果：与癌旁对照相比，脯氨酸代谢相关蛋白在HCC组织中显著上调。在急性营养应激下，外源性脯氨酸降低细胞ROS水平，减轻脂滴积聚，抑制HCC细胞过度自噬。脯氨酸恢复了营养剥夺诱导的肿瘤生长抑制，H2O2和雷帕霉素可以逆转这一抑制。结论：脯氨酸代谢通过减少ROS积累，从而抑制脂滴形成和自噬，维持营养限制下HCC细胞的存活。
{"title":"Proline metabolism supports the hepatocellular carcinoma cell survival by mitigating ROS generation under nutrient starvation","authors":"Jinglian Mo , Ziming Liu , Jialin Fu , Qichao Ge , Yiying Qu , Nan Ge , Yang Liu","doi":"10.1016/j.bbrc.2026.153392","DOIUrl":"10.1016/j.bbrc.2026.153392","url":null,"abstract":"<div><h3>Background</h3><div>Proline supplementation promotes proliferation and invasion in cancers under acute nutrient deprivation. However, its molecular basis in hepatocellular carcinoma is not fully understood yet. This study investigates how proline metabolism promotes hepatocellular carcinoma (HCC) cell proliferation and survival during nutrient starvation.</div></div><div><h3>Methods</h3><div>Proline metabolism-related protein expression in HCC versus paracarcinoma tissues was analyzed using The Cancer Genome Atlas database. HCC cell viability was quantified via Cell Counting Kit 8 assays, and colony formation capacity was evaluated. Intracellular ROS levels were measured by flow cytometry. Autophagic flux was assessed by GFP/mCherry fluorescence ratio and autophagy-related proteins by immunoblotting. Lipid droplet deposition was visualized using Hoechst 33342 and BODIPY 493/503 staining.</div></div><div><h3>Results</h3><div>Proline metabolism-related proteins were significantly upregulated in HCC tissues compared to paracarcinoma controls. Under acute nutrient stress, exogenous proline reduced cellular ROS levels, attenuated lipid droplet accumulation, and suppressed excessive autophagy in HCC cells. Proline rescued nutrient deprivation-induced tumor growth inhibition, which could be reversed by H<sub>2</sub>O<sub>2</sub> and rapamycin.</div></div><div><h3>Conclusion</h3><div>Proline metabolism sustains HCC cell survival during nutrient restriction by reducing ROS accumulation, thereby inhibiting lipid droplet formation and autophagy.</div></div>","PeriodicalId":8779,"journal":{"name":"Biochemical and biophysical research communications","volume":"805 ","pages":"Article 153392"},"PeriodicalIF":2.2,"publicationDate":"2026-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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