下调ClC-3通过抑制细胞自噬增强结直肠癌细胞对奥沙利铂化疗敏感性  

Down-regulation of ClC-3 enhances chemosensitivity of colorectal cancer cells to oxaliplatin by inhibiting autophagy

作　　者：段琼 陈斌辉 林义 Qiong Duan;Bin-Hui Chen;Yi Lin(School of Pharmacy,Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang Province,China;Department of Pharmacy,Enze Hospital,Taizhou Grace Medical Center(Group),Taizhou 317000,Zhejiang Province,China;Department of Pathology,First People’s Hospital of Wenling,Wenling 317500,Zhejiang Province,China)

机构地区：[1]浙江中医药大学药学院,浙江省杭州市310053 [2]台州恩泽医疗中心(集团)恩泽医院药剂科,浙江省台州市317000 [3]温岭市第一人民医院病理科,浙江省温岭市317500

出　　处：《世界华人消化杂志》2020年第24期1235-1242,共8页World Chinese Journal of Digestology

摘　　要：背景电压门控氯通道3(voltage-gated chloride channel 3,ClC-3)可参与调节卵巢癌、宫颈癌和肺癌的化疗敏感性的调节,而其在结直肠癌(colorectal cancer,CRC)的奥沙利铂化疗中的作用和机制尚不清楚.目的探究CRC细胞中ClC-3对奥沙利铂化疗敏感性的影响,以及细胞自噬在其中发挥的作用.方法用免疫组化染色观察ClC-3和微管相关蛋白1Ⅱ轻链3(microtubule-associated protein 1Ⅱlight chain 3,LC3-Ⅱ)在CRC耐药和敏感组织中的表达.用Western blot检测ClC-3和LC3-Ⅱ在CRC HT-29细胞以及奥沙利铂耐药细胞(HT-29/L-OHP)中的表达.用ClC-3 siRNA分别转染HT-29细胞和HT-29/L-OHP细胞后,CCK-8法检测细胞对奥沙利铂的敏感性变化;用Cyto-ID染色观察自噬点形成;用Western blot检测自噬相关蛋白Beclin1、LC3-Ⅰ、LC3-Ⅱ和p62的表达.用自噬激动剂雷帕霉素处理已转染ClC-3 siRNA的HT-29细胞和HT-29/L-OHP细胞后,用CCK-8法和Western blot分别检测细胞对奥沙利铂的敏感性变化以及Beclin1、LC3-Ⅰ、LC3-Ⅱ、p62和ClC-3蛋白的表达.结果CRC耐药组织以及HT-29/L-OHP细胞中LC3Ⅱ和ClC-3均呈现高表达.抑制ClC-3表达后,HT-29细胞和HT-29/L-OHP细胞对奥沙利铂的敏感性均增加,细胞的自噬水平降低.雷帕霉素能逆转抑制ClC-3对CRC细胞对奥沙利铂的增敏效应,但其对ClC-3的表达无影响.结论下调ClC-3可通过抑制细胞自噬来增强CRC细胞对奥沙利铂化疗的敏感性.BACKGROUND Voltage-gated chloride channel 3(ClC-3)can regulate the chemotherapy sensitivity of ovarian cancer,cervical cancer,and lung cancer,while its effect and mechanism in oxaliplatin therapy for colorectal cancer(CRC)are still unknown.AIM To investigate the effect of ClC-3 on oxaliplatin sensitivity and the role of autophagy in CRC cells.METHODS The expression of ClC-3 and microtubule-associated protein 1 light chain 3Ⅱ(LC3-Ⅱ)in the tissues of chemotherapy resistant and sensitive CRC was detected by immunohistochemical staining.The expression levels of ClC-3 and LC3-Ⅱin HT-29 and HT-29/L-OHP cells were detected by Western blot.After transfection of HT-29 and HT-29/L-OHP cells with ClC-3 siRNA,the sensitivity of the cells to oxaliplatin was detected by CCK-8 assay,the autophagy was detected by Cyto-ID staining,and the expression levels of autophagy-related proteins Beclin1,LC3-Ⅰ,LC3-Ⅱ,and p62 were detected by Western blot.The HT-29 and HT-29/L-OHP cells transfected with ClC-3 siRNA were subsequently treated with the autophagy agonist rapamycin,and then the sensitivity of the cells to oxaliplatin and the expression levels of Beclin1,LC3-Ⅰ,LC3-Ⅱ,p62,and ClC-3 were detected by CCK-8 assay and Western blot,respectively.RESULTS LC3-Ⅱand ClC-3 were highly expressed in tissues of chemo-resistant colorectal cancer and HT-29/L-OHP cells.After inhibition of ClC-3,the sensitivity of HT-29 cells and HT-29/L-OHP cells to oxaliplatin was both increased,and the autophagy level of the cells was decreased.Rapamycin reversed the sensitization of ClC-3 inhibition to oxaliplatin in colorectal cancer cells,but had no effect on the expression of ClC-3.CONCLUSION Down-regulation of ClC-3 can enhance the sensitivity of colorectal cancer cells to oxaliplatin chemotherapy by inhibiting autophagy.

关 键 词：CLC-3 细胞自噬 结直肠癌 奥沙利铂 化疗敏感性 

分 类 号：R735.34[医药卫生—肿瘤]