小干扰RNA技术沉默HLA-E基因表达对人乳腺癌细胞增殖与凋亡及裸鼠移植瘤生长的影响  

Effects of siRNA-mediated silencing of HLA-E gene on proliferation and apoptosis of human breast cancer cells and breast cancer transplanted tumors in nude mice

作　　者：吴振村 周艳[2] WU Zheng-cun;ZHOU Yan(不详;不详)

机构地区：[1]张家口市妇幼保健院检验科,河北张家口075000 [2]河北北方学院免疫教研室,河北张家口075000

出　　处：《广东医学》2022年第7期799-803,共5页Guangdong Medical Journal

基　　金：张家口市科技计划自筹经费项目(2021123D)。

摘　　要：目的探讨小干扰RNA(small interfering RNA,siRNA)技术沉默HLA-E基因表达对人乳腺癌细胞增殖与凋亡及对裸鼠移植瘤生长的影响。方法设计并合成靶向HLA-E基因的特异性siRNA及阴性对照siRNA序列,分别转染至人乳腺癌细胞MDA-MB-231,即为HLA-E siRNA组和阴性对照组,同时设立空白对照组(不加任何试剂)。Real-time PCR法和Western blot检测各组细胞HLA-E基因及蛋白表达;CCK-8法和流式细胞术检测各组乳腺癌细胞增殖和凋亡情况。将对数生长期MDA-MB-231细胞接种于裸鼠皮下,建立乳腺癌裸鼠移植瘤模型,随机分为3组:空白对照组、阴性对照组和HLA-E siRNA组,观察各组小鼠移植瘤生长情况。结果靶向HLA-E siRNA转染乳腺癌细胞MDA-MB-231后,HLA-E基因及蛋白表达明显降低;细胞增殖显著被抑制及细胞凋亡率显著增加(P<0.01);裸鼠体内实验结果显示,至接种30 d时,HLA-E siRNA组小鼠瘤组织体积显著低于空白对照组和阴性对照组(P<0.01)。结论siRNA技术沉默HLA-E基因表达能显著抑制乳腺癌细胞MDA-MB-231增殖,促进乳腺癌细胞凋亡,并抑制裸鼠移植瘤的生长,表明靶向沉默HLA-E基因表达可能为人乳腺癌基因治疗提供了新策略。Objective To analyze the effect of silencing HLA-E gene expression by small interfering RNA on the proliferation and apoptosis of breast cancer cells,and the growth of subcutaneous tumor of breast cancer cells in nude mice.Methods Specific and negative control siRNA sequences of HLA-E gene were designed and synthesized,then were specifically transfected into human breast cancer MDA-MB-231 cells;while the blank control group was established without any treatment.Real-time PCR and Western blot were used to detect HLA-E gene and protein expression.CCK-8 and flow cytometry were used to detect the proliferation and apoptosis of breast cancer cells.MDA-MB-231 cells were inoculated subcutaneously into nude mice to establish xenograft tumor model.The mice were randomly divided into three groups,blank control group,negative control group and HLA-E siRNA group.The growth of xenograft tumor in each group was observed.Results Compared with negative control group and blank control group,siRNA target against HLA-E could effectively down-regulate HLA-E gene and protein expression in MDA-MB-231 cells,and significantly inhibit the proliferation of cells,while significantly induce the apoptosis rate of the transfected breast cancer cells.In breast cancer bearing nude mice models,the average tumor volume in HLA-E siRNA group was significantly smaller than blank control group and negative control group(P<0.01).Conclusion Silencing HLA-E gene expression by siRNA could significantly inhibit the proliferation of MDA-MB-231 cells,promote the apoptosis of breast cancer cells and suppress the tumor growth in nude mice;indicating that targeted silencing HLA-E gene expression may provide a new strategy for gene therapy of human breast cancer.

关 键 词：乳腺癌 HLA-E 基因 小干扰RNA技术 增殖 凋亡 

分 类 号：R73.362[医药卫生—肿瘤] R392.11[医药卫生—临床医学]