HLA-E基因表达沉默对人乳腺癌细胞生物学行为的影响  

Effect of HLA-E gene silencing on biological behavior of breast cancer cells

作　　者：吴振村 周艳[2] WU Zhencun;ZHOU Yan(Maternal and Child Health Care Hospital of Zhangjiakou City,Zhangjiakou 075000;Hebei North University,Zhangjiakou 075000,Hebei,China)

机构地区：[1]张家口市妇幼保健院,河北张家口075000 [2]河北北方学院,河北张家口075000

出　　处：《癌变．畸变．突变》2023年第1期15-20,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：张家口市科技局重点研发计划项目资助(2021123D)。

摘　　要：目的:探讨HLA-E基因表达对人乳腺癌细胞增殖、凋亡、侵袭、迁移等生物学行为的影响,分析靶向HLA-E基因的小分子RNA干扰(siRNA)技术应用于乳腺癌治疗的可行性。方法:设计并合成靶向HLA-E基因的特异性siRNA序列,转染至人乳腺癌细胞MDA-MB-231,同时设立空白对照组、阴性对照组及脂质体组。实时荧光定量PCR(qPCR)和Western blot法分别检测各组乳腺癌细胞HLA-E mRNA及蛋白的表达;CCK-8法和流式细胞术检测各组乳腺癌细胞增殖率和凋亡率;Transwell实验检测各组乳腺癌细胞侵袭和迁移能力。结果:乳腺癌细胞转染HLA-E siRNA后,HLA-E mRNA及蛋白表达水平明显降低。与阴性对照组相比,HLA-E siRNA转染组乳腺癌细胞增殖率降低(P<0.01);凋亡率增加(P<0.01);侵袭迁移能力降低(P<0.01)。结论:靶向人乳腺癌细胞HLA-E基因的siRNA能有效抑制其基因表达,进而抑制细胞增殖,诱导细胞凋亡,降低乳腺癌细胞侵袭和迁移能力。OBJECTIVE:To investigate the expression of HLA-E gene in human breast cancer cells and its effect on rates of proliferation,apoptosis,invasion and migration of breast cancer cells,and analyze the feasibility of siRNA(small interfering RNA) silencing of HLA-E as gene therapy. METHODS:Specific siRNA sequences of HLA-E gene were designed and synthesized, and the sequences were transfected into human breast cancer cells MDA-MB-231 with liposome Vigofect. For comparisons,blank control,negative control and liposome groups were established. Quantitative real-time PCR and Western blot were used to detect HLA-E mRNA and protein expression. CCK-8 and flow cytometry were used to detect proliferation and apoptosis rates of the cancer cells. Transwell assay was used to detect the invasion and migration. RESULTS: After transfection with HLA-E siRNA, gene and protein expressions of HLA-E in the cancer cells were reduced significantly compared with the control groups. In addition, proliferation rates of the transfected cancer cells were significantly decreased(P<0.01),apoptosis rates were significantly increased(P<0.01) while invasion and migration were significantly decreased(P<0.01). CONCLUSION: siRNA targeting of HLA-E effectively suppressed its expression in the breast cancer cells together with inhibition of their proliferation, induction of apoptosis, and reduction of migration and invasion.

关 键 词：HLA-E基因 乳腺癌 小分子RNA干扰技术 基因治疗 

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