p52在结肠癌细胞对奥沙利铂耐药中的作用  

P52 confers resistance to oxaliplatin in oxaliplatin resistant colon cancer

作　　者：庞晓辉[1] 王朝杰[1] 崔勇霞[1] 周云[1] 

机构地区：[1]河南省人民医院郑州大学人民医院肿瘤科,郑州450003

出　　处：《中华实验外科杂志》2017年第9期1510-1512,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（U1204818）

摘　　要：目的观察p52在结肠癌细胞对奥沙利铂耐药中的作用。方法复苏结肠癌SW480、LoVo细胞、奥沙利铂处理细胞24 h，Western blot法检测SW480细胞中p52蛋白表达以研究奥沙利铂对p52蛋白的作用。采用大剂量冲击法诱导结肠癌耐奥沙利铂细胞，细胞计数试剂盒（CCK-8）实验检测其耐药性。Western blot检测原代耐药细胞p52表达，染色质免疫共沉淀（CHIP）实验检测p52能否转录调控B细胞淋巴瘤/白血病-2（bcl-2）。沉默耐药细胞中p52后，Western blot检测细胞内p52和bcl-2表达，膜联蛋白V/碘化丙锭（Annexin V/PI）染色、流式细胞术检测Annexin V（＋）/PI（＋）细胞的比例。SPSS 23.0统计软件进行统计分析。结果奥沙利铂上调结肠癌细胞中p52表达（t＝20.730，P＝0.002），p52蛋白在结肠癌耐奥沙利铂细胞中明显升高（t＝4.029，P＝0.006），差异均有统计学意义。CHIP实验结果显示p52蛋白结合到bcl-2基因启动子区域，转录调控bcl-2基因。沉默耐药细胞中p52后，bcl-2蛋白表达下调22.6%（t＝18.183，P＝0.003），流式细胞术提示实验组凋亡细胞比例为23.5%，比对照组升高了2.96倍（t＝6.783，P＝0.021）。 结论 p52通过通过上调bcl-2基因抑制细胞凋亡促使结肠癌细胞对奥沙利铂耐药。Objective To assess the role of p52 in resistance to oxaliplation in colon cancer. Methods Colon cancer cell lines SW480, LoVo and resistant cells （SW480/L-OHP, LoVo/L-OHP） were cultured. The inhibiting rates of SW480, LoVo cells and resistant cells by oxaliplatin were determined by cell counting kit-8 （CCK-8） assay. Western blotting was applied to assess induced p52 expression after cultured with oxaliplatin contained medium for 24 h in SW480 cells. We adopted chromatin immunoprecipitation （CHIP） to validate translational regulation of B cell lymphoma/leukemia-2 （bcl-2） by p52. SW480/L-OHP cells were cultured with oxaliplatin contained medium for 24 h after tranfected with siP52, then stained with Annexin V/propidium iodide （PI） in the detection of apoptosis cells by flow cytometry. Total protein was extracted and Western blotting was used to detect the expression of bcl-2, p52 after transfected with small interfering RNA （siRNA） targeting p52.Results Induced p52 expression was seen in SW480 cells. Translational regulation of bcl-2 by p52 was validated by CHIP assays. Down-regulation of bcl-2 was induced by sip52. The proportion of Annexin V/PI positive cells elevated in sip52 silenced SW480/L-OHP cells after treated with oxaliplatin for 24 h. Conclusion p52 confers resistance to oxaliplatin in oxaliplatin resistant colorectal cancer by inhibiting apoptosis.

关 键 词：结直肠癌 奥沙利铂 p52 

分 类 号：R735.35[医药卫生—肿瘤]