FANCF在三阴性乳腺癌细胞紫杉醇耐药中的作用及其机制  

The effects of FANCF on paclitaxel-resistant triple negative breast cancer cell

作　　者：马芸[1,2] 柴文英[2] 李姝墨 董坚[3] MA Yun1, 2, CHAI Wenying2, LI Shumo 2, DONG Jian3(1. Kunming Medical University, Kunming 650500, Yunnan, China; 2. Department of Breast Surgery, the First Affiliated Hospital of Kunming Medical University, Kunming 650032, Yunnan,China; 3. The Third Affiliated Hospital of Kunming Medical University, Kunming 650106, Yunnan, Chin)

机构地区：[1]昆明医科大学,云南昆明650500 [2]昆明医科大学第一附属医院乳腺外科,云南昆明650032 [3]昆明医科大学第三附属医院,云南昆明650106

出　　处：《中国肿瘤生物治疗杂志》2018年第3期240-245,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金项目(No.81660472);云南省卫生系统领军人才培养计划(No.L-201212);云南省细胞治疗技术转化医学重点实验室基金(No.2015DG034)~~

摘　　要：目的:建立三阴性乳腺癌紫杉醇耐药细胞株并考察FA/BRCA通路相关基因FANCF在该细胞株紫衫醇耐药中的作用。方法:以药物浓度递增法诱导三阴性乳腺癌细胞MDA-MB-231成为紫杉醇耐药细胞株,采用CCK8法检测细胞的耐药指数,流式细胞术检测细胞的生长周期,qRT-PCR检测FA/BRCA通路相关基因FANCF的表达;Western blotting法验证相关蛋白的表达。对MDA-MB-231敏感细胞和紫杉醇耐药细胞中FANCF的表达进行敲减,并在mRNA和蛋白水平进行敲减效果验证,以CCK8法检测紫杉醇对该两种细胞的IC_(50)。结果:MDA-MB-231细胞紫杉醇诱导3个月后的耐药指数为9.9,该细胞的G0/G1期细胞增多且S期细胞减少,细胞中FANCF mRNA和蛋白表达水平显著升高。FANCF敲低后无论MDA-MB-231还是MDA-MB-231/PTX细胞的凋亡增加,对紫杉醇敏感性显著升高(P<0.05或P<0.01)。结论:FANCF基因在乳腺癌紫杉醇耐药中具有重要作用,可能是乳腺癌治疗的一个潜在靶点。Objective： To establishpaclitaxel（PTX）-resistant human triple negative breast cancer cell line and to examine the expression profile of FA-related genes and FANCF, the correlation between the expression of FA-related genes, FANCF and PTX-resistance in breast cancer were further analyzed. Methods： PTX-resistant MDA-MB-231 cell line was established by means of long-term PTX-exposed culture. The sensitivity of the cells to paclitaxel was determined by the CCK8 assay. The cell cycle distribution was examined by flow cytometry after exposure to the paclitaxel. The expression of FA-related gene mRNA and FANCF protein were examined by using real time quantitative PCR and Western blotting. The expression of FANCF in the cells was reduced by RNAi interference technology and the effect of the RNAi was verified. Results： MDA-MB-231/PTX cell showed a 9.9-fold resistance to paclitaxel, indicating that the cell had acquired resistance to PTX. PTX treatment significantly induced G0/G1 arrest and the number of cells in phase S markedly decreased after exposure to PTX. The mRNA and protein expression of FANCF was significantly higher in PTX-resistant cell than that in PTX-sensitve parental cell,Knockdown of FANCF induced apoptosis in MDA-MB-231/PTX cell as well as in parental cell. FANCF knockdown increased the sensitivity of paclitaxel to both MDA-MB-231 and MDA-MB-231/PTX cells（P0.05 or P0.01）. Conclusion： FANCF played an important role in PTX resistance of the breast cancer cells and FANCF might be a target for therapy aimed at reversing chemoresistance.

关 键 词：乳腺癌 MDA-MB-231细胞 紫杉醇 耐药 FA/BRCA相关基因 FANCF基因 

分 类 号：R737.9[医药卫生—肿瘤] R730.5[医药卫生—临床医学]