耐药人胃癌细胞UHRF1基因的表达变化及其与多药耐药的关系  

Expression of UHRF1 gene in gastric cancer resistant cells and its relationship with multidrug resistance

作　　者：周林[1] 李伟[1] 韩东升[1] 张伟[1] 

机构地区：[1]中国人民解放军第八十八医院,山东泰安271000

出　　处：《山东医药》2016年第39期1-4,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81402337)

摘　　要：目的观察耐药人胃癌细胞泛素样含PHD和环指域1(UHRF1)基因的表达变化,并探讨其与胃癌细胞多药耐药的关系。方法取对数生长期人胃癌细胞SGC7901、耐长春新碱(VCR)的SGC7901(SGC7901/VCR)、耐阿霉素(ADR)的SGC7901(SGC7901/ADR),采用实时定量PCR法检测各细胞UHRF1 mRNA,采用Western blotting法检测各细胞UHRF1蛋白。取对数生长期SGC7901/VCR、SGC7901/ADR细胞分别转染抑制UHRF1表达的小干扰RNA(si UHRF1)(SGC7901/VCR-si UHRF1、SGC7901/ADR-si UHRF1)、无意义空白质粒(SGC7901/VCR-UHRF1、SGC7901/ADR-UHRF1),另设空白对照,不做任何处理。取对数生长期SGC7901细胞,分别转染过表达的UHRF1质粒(SGC7901-UHRF1)、无意义空白质粒(SGC7901-NC),另设空白对照。转染48 h收集以上细胞备用。采用MTT法检测SGC7901/VCR、SGC7901/ADR细胞各干扰组和SGC7901细胞各干扰组对化疗药物[(VCR、ADR、5-氟尿嘧啶(5-FU)和顺氨氯铂(CDDP)]的药物敏感性,计算半数抑制浓度(IC_(50));取SGC7901/VCR、SGC7901/ADR细胞各干扰组和SGC7901空白对照,加入5-FU共培养,采用流式细胞仪观察细胞凋亡情况。结果 SGC7901细胞UHRF1 mRNA及蛋白的相对表达量均低于SGC7901/VCR、SGC7901/ADR细胞,P均<0.05。促进UHRF1表达可升高SGC7901细胞对VCR、ADR、5-FU、CDDP的IC_(50),降低细胞凋亡率,P均<0.05。抑制UHRF1表达可降低VCR、ADR、5-FU、CDDP对SGC7901/VCR和SGC7901/ADR细胞的IC_(50),升高细胞凋亡率,P均<0.05。结论耐药人胃癌细胞中存在UHRF1高表达。UHRF1高表达可降低胃癌细胞的药物敏感性、提高细胞多药耐药性,可能与UHRF1抑制胃癌细胞凋亡有关。Objective To observe the expression changes of ubiquitin-like with PHD and ring finger domain 1（ UHRF1） in gastric cancer drug-resistance cells,and to explore the relationship with gastric cancer multidrug resistance.Methods We examined UHRF1 mRNA and protein expression in GC multidrug-resistant cell lines SGC7901/VCR and SGC7901/ADR and their parental cell line SGC7901 by RT-PCR and Western blotting. Small interfering RNA（ si UHRF1）was transfected into SGC7901/VCR and SGC7901/ADR cells（ SGC7901/VCR-si UHRF1,SGC7901/ADR-si UHRF1）,respectively; and blank control group without any treatment was set. UHRF1 plasmid with overexpression（ SGC7901-UHRF1） and empty blank plasmid（ SGC7901-NC） were transfected into SGC7901 cells,and blank control group was also established. After 48-hour transfection,the cells were collected for further use. MTT assay was conducted to detect the GC cell line sensitivity to several chemotherapy drugs,including VCR,ADR,5-FU and CDDP,and IC_（50） was calculated. 5-FU was co-cultured with gastric cancer cell lines and the apoptosis was observed by flow cytometry（ FCM）. Results The relative mRNA and protein expression of UHRF1 in SGC7901 cell line was down-regulated as compared with that of SGC7901/VCR and SGC7901/ADR cells（ all P〈0. 05）. Promoting the expression of UHRF1 could increase the IC_（50） of SGC7901cells to VCR,ADR,5-FU and CDDP and the apoptosis was decreased（ all P〈0. 05）. Silencing UHRF1 expression could reduce the IC_（50） of SGC7901/VCR and SGC7901/ADR cells to VCR,ADR,5-FU and CDDP,and the apoptosis rate was increased（ all P〈0. 05）. Conclusions UHRF1 is highly expressed in gastric cancer drug-resistance cell lines,and the high expression of UHRF1 may decrease drug sensitivity and improve multidrug resistance of gastric cancer,which may be related to the inhibition of UHRF1 on the apoptosis of gastric cancer cells.

关 键 词：胃癌 泛素样含PHD和环指域1 多药耐药 药物敏感性 半数抑制浓度 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]