白藜芦醇促进膀胱癌细胞对TRAIL凋亡诱导作用的研究
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摘要
目的探讨白藜芦醇对膀胱癌细胞的作用及其与TRAIL联合对膀胱癌细胞的凋亡诱导作用及其可能的机制。方法MTT法检测白藜芦醇或/和TRAIL对膀胱癌细胞5637和T24体外生长能力的影响。流式细胞术测定白藜芦醇和/或TRAIL对细胞凋亡和线粒体跨膜电位的影响。Western blot检测白藜芦醇对死亡受体(DR)4、5与细胞凋亡相关蛋白表达水平的变化。结果 MTT证实白藜芦醇可抑制膀胱癌细胞体外生长能力,且呈剂量与时间依赖性。单独应用TRAIL未能抑制膀胱癌细胞的生长,但与白藜芦醇联合应用时可显著抑制细胞的增殖速度。白藜芦醇能够诱导膀胱癌细胞发生凋亡,而TRAIL则不能诱导细胞凋亡,但联合应用白藜芦醇和TRAIL可诱导膀胱癌细胞发生显著的凋亡。此外,在白藜芦醇或联合应用白藜芦醇与TRAIL可诱导肿瘤细胞丢失线粒体膜电位(ΔΨm),而单独应用TRAIL则未能诱导线粒体膜电位的丢失。Western blot结果表明,白藜芦醇可上调DR4和DR5的表达,并下调Bcl-2与survivin的表达。结论白藜芦醇可抑制膀胱癌5637和T24细胞增殖并诱导细胞凋亡。此外,白藜芦醇可以提高膀胱癌细胞对TRAIL诱导的细胞凋亡作用。单独应用白藜芦醇或与TRAIL联合应用可能成为临床膀胱癌治疗的新策略。
Objective To investigate the effects of resveratrol on bladder cancer cells,the interactive role with TRAIL on the apoptosis of bladder cancer cells,and the possible mechanisms.Methods The effects of resveratrol or/and TRAIL on the growth of 5637 and T24 cells were detected with MTT.The apoptosis and mitochondrial transmembrane potential(ΔΨm)after treatment of resveratrol or/and TRAIL were determined with flow cytometry.Death receptor(DR)4 and5,and apoptosis associated proteins were detected with Western blot.Results Resveratrol inhibited cell viability in a dose and time-dependent manner.TRAIL alone failed to inhibit cell viability in5637 and T24 cells,whereas it could inhibit cell proliferation in the presence of resveratrol.Resveratrol alone was able to induce apoptosis in both cells,whereas TRAIL failed to induce apoptosis.Notably,combination of resveratrol and TRAIL resulted in significant apoptosis in tumor cells.Furthermore,a significant loss ofΔΨm was obtained in resveratrol alone and combination of resveratrol and TRAIL treated cells,and TRAIL alone failed to triggerΔΨm collapse.Resveratrol upregulated the expressions of DR4 and DR5 while downregulated the expressions of Bcl-2 and survivin.Conclusion Resveratrol inhibits the proliferation of 5637 and T24 cells in a time and dose-dependent manner and induces cell apoptosis.In addition,resveratrol can sensitize bladder cancer cells to TRAIL-induced apoptosis.The capability of resveratrol to inhibit tumor growth and enhance the therapeutic potential of TRAIL suggests that resveratrol alone or in combination with TRAIL can be used for the management of bladder cancer.
Objective To investigate the effects of resveratrol on bladder cancer cells,the interactive role with TRAIL on the apoptosis of bladder cancer cells,and the possible mechanisms.Methods The effects of resveratrol or/and TRAIL on the growth of 5637 and T24 cells were detected with MTT.The apoptosis and mitochondrial transmembrane potential(ΔΨm)after treatment of resveratrol or/and TRAIL were determined with flow cytometry.Death receptor(DR)4 and5,and apoptosis associated proteins were detected with Western blot.Results Resveratrol inhibited cell viability in a dose and time-dependent manner.TRAIL alone failed to inhibit cell viability in5637 and T24 cells,whereas it could inhibit cell proliferation in the presence of resveratrol.Resveratrol alone was able to induce apoptosis in both cells,whereas TRAIL failed to induce apoptosis.Notably,combination of resveratrol and TRAIL resulted in significant apoptosis in tumor cells.Furthermore,a significant loss ofΔΨm was obtained in resveratrol alone and combination of resveratrol and TRAIL treated cells,and TRAIL alone failed to triggerΔΨm collapse.Resveratrol upregulated the expressions of DR4 and DR5 while downregulated the expressions of Bcl-2 and survivin.Conclusion Resveratrol inhibits the proliferation of 5637 and T24 cells in a time and dose-dependent manner and induces cell apoptosis.In addition,resveratrol can sensitize bladder cancer cells to TRAIL-induced apoptosis.The capability of resveratrol to inhibit tumor growth and enhance the therapeutic potential of TRAIL suggests that resveratrol alone or in combination with TRAIL can be used for the management of bladder cancer.
引文
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[8]SINGH CK,NDIAYE MA,AHMAD N.Resveratrol and cancer:Challenges for clinical translation[J].Biochim Biophys Acta,2015,1852(6):1178-1185.
[9]XU Q,ZONG L,CHEN X,et al.Resveratrol in the treatment of pancreatic cancer[J].Ann N Y Acad Sci,2015,1348(1):10-19.
[10]SPROUSE AA,HERBERT BS.Resveratrol augments paclitaxel treatment in MDA-MB-231and paclitaxel-resistant MDA-MB-231breast cancer cells[J].Anticancer Res,2014,34(10):5363-5374.
[11]LIU YZ,WU K,HUANG J,et al.The PTEN/PI3K/Akt and Wnt/β-catenin signaling pathways are involved in the inhibitory effect of resveratrol on human colon cancer cell proliferation[J].Int J Oncol,2014,45(1):104-112.
[12]WANG F,LIN J,XU R.The molecular mechanisms of TRAILresistance in cancer cells:help in designing new drugs[J].Curr Pharm Des,2014,20(42):6714-6722.
[13]LI Y,JIN X,LI J,et al.Expression of TRAIL,DR4,and DR5in bladder cancer:correlation with response to adjuvant therapy and implications of prognosis[J].Urology,2012,79(4):968.e7-e15.
[2]STUCKEY DW,SHAH K.TRAIL on trial:preclinical advances in cancer therapy[J].Trends Mol Med,2013,19(11):685-694.
[3]LEAHOMSCHI S,MOLINSKY J,KLANOVA M,et al.Multilevel disruption of the extrinsic apoptotic pathway mediates resistance of leukemia cells to TNF-related apoptosis-inducing ligand(TRAIL)[J].Neoplasma,2013,60(2):223-231.
[4]GUIHO R,BITEAU K,HEYMANN D,et al.TRAIL-based therapy in pediatric bone tumors:how to overcome resistance[J].Future Oncol,2015,11(3):535-542.
[5]HSIEH TC,WU JM.Resveratrol:Biological and pharmaceutical properties as anticancer molecule[J].Biofactors,2010,36(5):360-369.
[6]OIKONOMOU E,PINTZAS A.The TRAIL of oncogenes to apoptosis[J].Biofactors,2013,39(4):343-354.
[7]ZHUANG H,JIANG W,ZHANG X,et al.Suppression of HSP70expression sensitizes NSCLC cell lines to TRAIL-induced apoptosis by upregulating DR4and DR5and downregulating cFLIP-L expressions[J].J Mol Med(Berl).2013,91(2):219-235.
[8]SINGH CK,NDIAYE MA,AHMAD N.Resveratrol and cancer:Challenges for clinical translation[J].Biochim Biophys Acta,2015,1852(6):1178-1185.
[9]XU Q,ZONG L,CHEN X,et al.Resveratrol in the treatment of pancreatic cancer[J].Ann N Y Acad Sci,2015,1348(1):10-19.
[10]SPROUSE AA,HERBERT BS.Resveratrol augments paclitaxel treatment in MDA-MB-231and paclitaxel-resistant MDA-MB-231breast cancer cells[J].Anticancer Res,2014,34(10):5363-5374.
[11]LIU YZ,WU K,HUANG J,et al.The PTEN/PI3K/Akt and Wnt/β-catenin signaling pathways are involved in the inhibitory effect of resveratrol on human colon cancer cell proliferation[J].Int J Oncol,2014,45(1):104-112.
[12]WANG F,LIN J,XU R.The molecular mechanisms of TRAILresistance in cancer cells:help in designing new drugs[J].Curr Pharm Des,2014,20(42):6714-6722.
[13]LI Y,JIN X,LI J,et al.Expression of TRAIL,DR4,and DR5in bladder cancer:correlation with response to adjuvant therapy and implications of prognosis[J].Urology,2012,79(4):968.e7-e15.