Objectives:Fractionated radiotherapy represents a standardized and widely adopted treatment modality for cancermanagement,with approximately 40% of non-small cell lung cancer(NSCLC)patients receiving it.However,repeat...Objectives:Fractionated radiotherapy represents a standardized and widely adopted treatment modality for cancermanagement,with approximately 40% of non-small cell lung cancer(NSCLC)patients receiving it.However,repeated irradiationmay induce radioresistance in cancer cells,reducing treatment effectiveness and raising recurrence risk.The long noncoding RNA CRYBG3(lncRNA CRYBG3),which is upregulated in lung cancer cells after Xray irradiation,contributes to the radioresistance of NSCLC cells by promoting wild-type p53 protein degradation.This study aims to elucidate the mechanism of fractionated irradiation-induced radioresistance,in which lncRNA CRYBG3 regulates radiation-induced mitochondrial damage and reactive oxygen species(ROS)generation through the p53 downstream signaling pathway.Methods:To investigate the critical roles of lncRNA CRYBG3 in mediating radioresistance induced by fractionated irradiation in NSCLC,we established radioresistant NSCLC cells by irradiating the A549 and H460 cell lines with 60 Gy of X-rays in 12 fractions,and named the radioresistant cells A549R and H460R,respectively.Lentiviral vectors were used to deliver short hairpin RNA(shRNA)into cells to knock down lncRNA CRYBG3,thereby investigating its contribution to adaptive radioresistance in A549R and H460R cells.All cells were irradiated with 4 Gy of X-rays,and subsequent analyses were conducted to evaluate mitochondrial damage,ROS generation,apoptosis,and the expression of oxidative stress-related proteins.Results:Increased expression levels of lncRNA CRYBG3 were positively associated with the acquisition of radioresistance in NSCLC cells.Additionally,suppressing lncRNA CRYBG3 increased mitochondrial damage and promoted radiation-induced apoptosis in radioresistant NSCLC cells.Mechanistically,the downregulation of lncRNA CRYBG3 led to increased p53 levels,resulting in decreased expression of heat shock factor 1(HSF1)and tumor necrosis factor receptor-associated protein 1(TRAP1),as well as enhanced radiation-induced mitochondrial oxidative damage and apoptosis.Conclusion:The results indicate that lncRNA CRYBG3 plays a regulatory role in adaptive radioresistance in NSCLC cells through the p53/HSF1/TRAP1 axis.Therefore,targeting lncRNA CRYBG3 could potentially improve the efficacy of fractionated radiotherapy in NSCLC.展开更多
背景:肿瘤坏死因子受体相关蛋白1(TRAP1)是线粒体特异性热休克蛋白90家族的同源基因。大量研究表明其过量表达与多种癌症的发生密切相关。但其在喉鳞癌发生中的作用及作用机制目前还不清楚。目的:探讨RNA干扰能否靶向抑制TRAP1过表达以...背景:肿瘤坏死因子受体相关蛋白1(TRAP1)是线粒体特异性热休克蛋白90家族的同源基因。大量研究表明其过量表达与多种癌症的发生密切相关。但其在喉鳞癌发生中的作用及作用机制目前还不清楚。目的:探讨RNA干扰能否靶向抑制TRAP1过表达以及对人喉癌干细胞增殖和凋亡的作用效果。方法:采用免疫磁珠技术从人喉癌Hep-2细胞系中分选出CD133^+CD44^+喉癌干细胞。设计及合成TRAP1的sh RNA序列,Lipofectamine^(TM) 2000转染CD133^+CD44^+喉癌干细胞。CCK-8增殖实验、集落形成实验、流式细胞术检测干扰TRAP1基因表达对CD133^+CD44^+喉癌干细胞增殖和凋亡的影响,采用分光光度法检测细胞内caspase-3,caspase-8,caspase-9活性。结果与结论:(1)TRAP1 sh RNA转染的CD133^+CD44^+喉癌干细胞内TRAP1基因和蛋白表达明显下调(P<0.01);(2)与空白对照组和转染空质粒阴性对照组比较,TRAP1表达下调抑制了CD133^+CD44^+喉癌干细胞增殖和集落形成能力(P<0.05);(3)与空白对照组和转染空质粒阴性对照组比较,TRAP1表达下调增加CD133^+CD44^+喉癌干细胞凋亡率(P<0.05);(4)TRAP1 sh RNA介导细胞凋亡与caspase-3,caspase-8和caspase-9激活有关;(5)结果提示RNA干扰TRAP1表达可抑制CD133^+CD44^+喉癌干细胞生长并促进细胞凋亡。TRAP1可能为喉鳞癌治疗的基因靶点。展开更多
Tumor necrosis factor receptor-associated protein 1 （TRAP1）, an important member of mitochondrial heat shock protein 90 family, is involved in multiple biological processes in several types of tumors. However, its p...Tumor necrosis factor receptor-associated protein 1 （TRAP1）, an important member of mitochondrial heat shock protein 90 family, is involved in multiple biological processes in several types of tumors. However, its pathological role in esophageal squamous cell cancer （ESCC） remains unknown. Herein, we demonstrated the clinical value of TRAP1, and its role in apoptosis and motility in ESCC. The clinical potential of TRAP1 was investigated through immunohistochemical analysis in 328 ESCC samples, which revealed that strong TRAP1 expression was associated with increased risk of lymph node metastasis, while high TRAP1 expression correlated with poor prognosis. Expression of TRAP1 was found to be an independent prognostic factor for patients with ESCC. Additionally, the upregulation of TRAP1 antagonized cisplatin-induced apoptosis while its downregulation sensitized cells to cisplatin-induced apoptosis. As revealed by the transwell assay, TRAP1 overexpression promoted cellular migration and invasion as compared to the control groups. In contrast, silencing of endogenous TRAP1 expression attenuated the ability of migration and invasion. Finally, the molecular mechanism investigated in the present study demonstrated that TRAP1-mediated migration and invasion occurred through STAT3/MMP2 signaling pathway. In conclusion, TRAP1 may be considered as a molecular predictive marker for prognosis and a novel molecular candidate for therapeutic target in ESCC.展开更多
基金supported by the National Natural Science Foundation of China(No.12205215)Suzhou Fundamental Research Project(No.SJC2023001)+2 种基金Basic Research Program of Nantong Science and Technology Bureau(No.JC12022103)College Student Innovation Training Program Project of Soochow University(No.202410285101Z)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD).
文摘Objectives:Fractionated radiotherapy represents a standardized and widely adopted treatment modality for cancermanagement,with approximately 40% of non-small cell lung cancer(NSCLC)patients receiving it.However,repeated irradiationmay induce radioresistance in cancer cells,reducing treatment effectiveness and raising recurrence risk.The long noncoding RNA CRYBG3(lncRNA CRYBG3),which is upregulated in lung cancer cells after Xray irradiation,contributes to the radioresistance of NSCLC cells by promoting wild-type p53 protein degradation.This study aims to elucidate the mechanism of fractionated irradiation-induced radioresistance,in which lncRNA CRYBG3 regulates radiation-induced mitochondrial damage and reactive oxygen species(ROS)generation through the p53 downstream signaling pathway.Methods:To investigate the critical roles of lncRNA CRYBG3 in mediating radioresistance induced by fractionated irradiation in NSCLC,we established radioresistant NSCLC cells by irradiating the A549 and H460 cell lines with 60 Gy of X-rays in 12 fractions,and named the radioresistant cells A549R and H460R,respectively.Lentiviral vectors were used to deliver short hairpin RNA(shRNA)into cells to knock down lncRNA CRYBG3,thereby investigating its contribution to adaptive radioresistance in A549R and H460R cells.All cells were irradiated with 4 Gy of X-rays,and subsequent analyses were conducted to evaluate mitochondrial damage,ROS generation,apoptosis,and the expression of oxidative stress-related proteins.Results:Increased expression levels of lncRNA CRYBG3 were positively associated with the acquisition of radioresistance in NSCLC cells.Additionally,suppressing lncRNA CRYBG3 increased mitochondrial damage and promoted radiation-induced apoptosis in radioresistant NSCLC cells.Mechanistically,the downregulation of lncRNA CRYBG3 led to increased p53 levels,resulting in decreased expression of heat shock factor 1(HSF1)and tumor necrosis factor receptor-associated protein 1(TRAP1),as well as enhanced radiation-induced mitochondrial oxidative damage and apoptosis.Conclusion:The results indicate that lncRNA CRYBG3 plays a regulatory role in adaptive radioresistance in NSCLC cells through the p53/HSF1/TRAP1 axis.Therefore,targeting lncRNA CRYBG3 could potentially improve the efficacy of fractionated radiotherapy in NSCLC.
文摘背景:肿瘤坏死因子受体相关蛋白1(TRAP1)是线粒体特异性热休克蛋白90家族的同源基因。大量研究表明其过量表达与多种癌症的发生密切相关。但其在喉鳞癌发生中的作用及作用机制目前还不清楚。目的:探讨RNA干扰能否靶向抑制TRAP1过表达以及对人喉癌干细胞增殖和凋亡的作用效果。方法:采用免疫磁珠技术从人喉癌Hep-2细胞系中分选出CD133^+CD44^+喉癌干细胞。设计及合成TRAP1的sh RNA序列,Lipofectamine^(TM) 2000转染CD133^+CD44^+喉癌干细胞。CCK-8增殖实验、集落形成实验、流式细胞术检测干扰TRAP1基因表达对CD133^+CD44^+喉癌干细胞增殖和凋亡的影响,采用分光光度法检测细胞内caspase-3,caspase-8,caspase-9活性。结果与结论:(1)TRAP1 sh RNA转染的CD133^+CD44^+喉癌干细胞内TRAP1基因和蛋白表达明显下调(P<0.01);(2)与空白对照组和转染空质粒阴性对照组比较,TRAP1表达下调抑制了CD133^+CD44^+喉癌干细胞增殖和集落形成能力(P<0.05);(3)与空白对照组和转染空质粒阴性对照组比较,TRAP1表达下调增加CD133^+CD44^+喉癌干细胞凋亡率(P<0.05);(4)TRAP1 sh RNA介导细胞凋亡与caspase-3,caspase-8和caspase-9激活有关;(5)结果提示RNA干扰TRAP1表达可抑制CD133^+CD44^+喉癌干细胞生长并促进细胞凋亡。TRAP1可能为喉鳞癌治疗的基因靶点。
基金supported by fundings from the National Basic Research Program of China (Nos. 2011CB910801 and 2012CB967003)the National Natural Science Foundation of China (No. 81071633)
文摘Tumor necrosis factor receptor-associated protein 1 （TRAP1）, an important member of mitochondrial heat shock protein 90 family, is involved in multiple biological processes in several types of tumors. However, its pathological role in esophageal squamous cell cancer （ESCC） remains unknown. Herein, we demonstrated the clinical value of TRAP1, and its role in apoptosis and motility in ESCC. The clinical potential of TRAP1 was investigated through immunohistochemical analysis in 328 ESCC samples, which revealed that strong TRAP1 expression was associated with increased risk of lymph node metastasis, while high TRAP1 expression correlated with poor prognosis. Expression of TRAP1 was found to be an independent prognostic factor for patients with ESCC. Additionally, the upregulation of TRAP1 antagonized cisplatin-induced apoptosis while its downregulation sensitized cells to cisplatin-induced apoptosis. As revealed by the transwell assay, TRAP1 overexpression promoted cellular migration and invasion as compared to the control groups. In contrast, silencing of endogenous TRAP1 expression attenuated the ability of migration and invasion. Finally, the molecular mechanism investigated in the present study demonstrated that TRAP1-mediated migration and invasion occurred through STAT3/MMP2 signaling pathway. In conclusion, TRAP1 may be considered as a molecular predictive marker for prognosis and a novel molecular candidate for therapeutic target in ESCC.
