HLA-E基因表达沉默对人乳腺癌细胞生物学行为的影响

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。

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

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.