UbcH10基因沉默联合多柔比星抑制耐药乳腺癌细胞体内成瘤实验  

UbcH10 gene silencing combined with doxorubicin inhibits in vivo tumor formation of drug-resistant breast cancer cells

作　　者：汪成[1] 余燕民[1] 陆永伟[1] 单鸣[1] 徐明[2] 包家林 

机构地区：[1]上海市黄浦区中心医院乳腺外科,上海200002 [2]上海市黄浦区中心医院病理科,上海200002

出　　处：《第二军医大学学报》2015年第4期418-422,共5页Academic Journal of Second Military Medical University

基　　金：上海市医学重点专科建设项目(ZK2012A13)~~

摘　　要：目的观察UbcH10基因沉默对多柔比星抑制耐药乳腺癌细胞株MCF-7/ADR体内成瘤作用的影响。方法采用慢病毒系统在MCF-7/ADR细胞中进行UbcH10基因沉默。将基因沉默后的肿瘤细胞及对照肿瘤细胞接种至裸鼠皮下,通过裸鼠尾静脉连续给予多柔比星或生理盐水2周并停药1周,测量肿瘤体积,分析UbcH10沉默对多柔比星抑制MCF-7/ADR体内成瘤作用的影响。采用蛋白质印迹分析检测瘤体中的UbcH10及BCL-2蛋白含量,分析UbcH10与肿瘤细胞化疗敏感性之间的调控关系。结果通过慢病毒实验系统成功建立UbcH10基因沉默细胞株MCF-7/ADR UbcH10-RNAi。经过3周给药处理,多柔比星组肿瘤体积与对照组比较差异无统计学意义,肿瘤抑制率为4.16%;基因沉默+多柔比星组肿瘤体积与对照组差异有统计学意义(P<0.05),肿瘤抑制率为41.8%。瘤体内蛋白含量检测结果显示,对照组、多柔比星组、基因沉默+多柔比星组UbcH10蛋白表达量分别为0.81±0.16、0.78±0.12、0.18±0.04,基因沉默+多柔比星组其余两组比较差异均有统计学意义(P<0.05);BCL-2蛋白表达量组间差异与UbcH10一致,二者之间具有正相关性。结论UbcH10基因沉默可以显著增强多柔比星抑制耐药乳腺癌细胞MCF-7/ADR的体内成瘤作用。Objective To observe the effect of the UbcH10 gene silencing on inhibition effect of doxorubicin against in vivo tumor formation of drug-resistant breast cancer MCF-7/ADR cells.Methods An MCF-7/ADR-UbcH10-RNAi cell line with UbcH10 gene silenced was established.Then MCF-7/ADR-UbcH10-RNAi cells and the control cells at logarithmic phase were inoculated into nude mice to establish the subcutaneous tumor model.Doxorubicin or normal saline was administered for a consecutive of two weeks,and then one week later the tumor volumes were determined to analyze the effects of UbcH10 gene silencing on inhibition of tumor formation by doxorubicin.Meanwhile,Western blotting analysis was used to examine the protein expression of UbcH10 and BCL-2;the relationship between UbcH10 and chemosensitivity of tumor cells was analyzed.Results The MCF-7/ADR-UbcH10-RNAi cell line with UbcH10 gene silenced was successfully established with the lentiviral experimental system.The nude mice tumor models were established three weeks after subcutaneous inoculation of tumor cells.The tumor inhibitory rate was 4.16% in the doxorubicin group,which was not significantly different from the control group（P〉0.05）;while the tumor inhibitory rate was 41.8%in UbcH10-RNAi＋doxorubicin group,which was significantly higher than that in the control group（P〈0.05）.The results of Western blotting analysis showed that the expression levels of UbcH10 in MCF-7/ADR group,MCF-7/ADR＋doxorubicin group,and MCF-7/ADR-UbcH10-RNAi＋doxorubicin group were0.81±0.16,0.78±0.12,and 0.18±0.04,respectively,with the latter being significantly lower than the former two（P〈0.05）;BCL-2protein levels in tumors were consistent with those of UbcH10.Conclusion UbcH10 gene silencing can markedly enhance the in vivo sensitivity of drug resistant breast cancer cells to Doxorubicin.

关 键 词：乳腺肿瘤 MCF-7/ADR 基因沉默 UBCH10 肿瘤抗药性 体内成瘤 

分 类 号：R737.9[医药卫生—肿瘤]