化疗联合自噬抑制剂对结肠癌细胞钙网蛋白膜表达的影响_彭瑞清.pdf

Med J Chin PLA, Vol. 41, No. 4, April 1, 2016 　273 论　著 化疗联合自噬抑制剂对结肠癌细胞钙网蛋白膜表达的影响 彭瑞清，李丹丹，丁娅，张星，张晓实，伍小军 [摘要]　目的　研究化疗药物联合自噬抑制剂对结肠癌细胞凋亡以及钙网蛋白(CRT)膜表达的影响。方法　以结 肠癌细胞HCT116作为研究对象，采用MTT法检测化疗药物奥沙利铂、5-Fu和SN38对细胞的IC50值，流式细胞术检测 化疗药物处理前后的细胞凋亡率和CRT膜表达比例的变化，免疫荧光法观察药物处理前后结肠癌细胞CRT表达部位的 变化，Western blotting检测药物处理对HCT116细胞自噬的影响，并用流式细胞术检测药物联合自噬抑制剂氯喹(CQ) 处理前及处理12h后细胞凋亡率和CRT膜表达比例的变化。结果　化疗药物奥沙利铂、5-Fu和SN38处理HCT116细胞 12h后，细胞凋亡率和CRT膜表达比例均较对照组有所上升，但差异均无统计学意义。免疫荧光实验提示奥沙利铂处 理后，CRT由细胞质转位至细胞膜。Western blotting结果提示3种化疗药物均可诱导HCT116细胞产生自噬，联合CQ后 其诱导自噬的作用受到抑制。化疗药物联合CQ处理HCT116细胞后，细胞凋亡率和CRT膜表达比例均较未用药物处 理和单纯化疗药物诱导组增高；奥沙利铂联合CQ处理后CRT膜表达比例升高，与药物处理前比较差异有统计学意义 (P=0.027)。结论　化疗药物奥沙利铂联合自噬抑制剂CQ能使结肠癌HCT116细胞凋亡增多，CRT膜表达比例上升。 [关键词]　钙网蛋白；免疫原性；化疗；自噬 [中图分类号]　R733.712　　　　[文献标志码]　A　　　　[文章编号]　0577-7402(2016)04-0273-06 [DOI]　10.11855/j.issn.0577-7402.2016.04.03 Influences of combination of chemotherapy and autophagy inhibitor on the calreticulin expression in colon cancer cells PENG Rui-qing1, LI Dan-dan1, DING Ya1, ZHANG Xing1, ZHANG Xiao-shi1, WU Xiao-jun2* 1Biotherapy Center, 2Department of Colorectal Surgery, Cancer Center, Sun Yat-sen University, Guangzhou 510060, China *Corresponding author, E-mail: wuxj@sysucc.org.cn This work was supported by the National Natural Science Foundation of China , and the Medical Research Foundation of Guangdong Province (B2011112) [Abstract]　Objective　To investigate the influence of chemotherapy combined with autophagy inhibitor on apoptosis and calreticulin (CRT) expression on colonic cancer cells. Methods　The colon cancer cells HCT116 were taken as the target in the present study. The inhibition rates (IC50) of chemotherapeutics oxaliplatin, 5-Fu and SN-38 were assessed by MTT assay. The changes in CRT expression on the membrane of HCT116 and apoptosis were determined with flow cytometry before and after treatment with chemotherapeutics. CRT locatio