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目的:探讨siRNA(small interference RNA)干扰趋化因子受体CXCR4和CXCR7对子宫内膜癌HEC-1-A细胞生物学行为的影响。方法:以LipofectamineTM2000脂质体为载体将siRNA转染至人子宫内膜癌HEC-1-A细胞株,以半定量RTPCR和Western blotting验证siRNA对CXCR4和CXCR7的干扰效果,以CCK-8法、流式细胞术和Transwell法分别检测CXCR4和CXCR7基因沉默对HEC-1-A细胞周期、增殖和侵袭能力的影响;结果:HEC-1-A细胞高表达CXCR4和CXCR7,合成的CXCR4-siRNA、CXCR7-siRNA、CXCR4/CXCR7-siRNA均可分别干扰HEC-1-A细胞内CXCR4、CXCR7、CXCR4/CXCR7基因或蛋白的表达;CXCR4和CXCR7基因的单独沉默或联合沉默均可导致HEC-1-A细胞增殖和侵袭能力的显著下降(P<0.05),同时引发HEC-1-A细胞周期出现S期阻滞。结论:趋化因子受体CXCR4和CXCR7在HEC-1-A细胞的增殖和侵袭行为中发挥重要作用,CXCR4和CXCR7途径的抑制有望成为子宫内膜癌治疗的新靶点。
AIM: To investigate the effect of siRNA on the biological behavior of endometrial carcinoma HEC-1-A cells by interfering with the chemokine receptor CXCR4 and CXCR7. METHODS: siRNA was transfected into human endometrial carcinoma cell line HEC-1-A by LipofectamineTM 2000 liposome. The interference effects of siRNA on CXCR4 and CXCR7 were verified by semi-quantitative RT-PCR and Western blotting. The CCK-8, Flow cytometry and Transwell method were used to detect the effect of CXCR4 and CXCR7 gene silencing on the cell cycle, proliferation and invasion of HEC-1-A cells. Results: CXCR4 and CXCR7 were highly expressed in HEC-1-A cells, The expression of CXCR4, CXCR7 and CXCR4 / CXCR7 genes or proteins in HEC-1-A cells can be interfered by both CXCR7-siRNA and CXCR4 / CXCR7-siRNA. A cell proliferation and invasive ability was significantly decreased (P <0.05), while triggering S phase HEC-1-A cell cycle arrest. CONCLUSION: The chemokine receptors CXCR4 and CXCR7 play an important role in the proliferation and invasion of HEC-1-A cells. The inhibition of CXCR4 and CXCR7 pathways is expected to become a new target for the treatment of endometrial cancer.