RNA干扰CXCR4和CXCR7对子宫内膜癌HEC-1-A细胞生物学行为的影响

Effects of CXCR4 and CXCR7 downregulation by RNA interference on the biological behaviors of endometrial carcinoma HEC-1-A cells

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。

Objective: To investigate the effects of chemokine receptors CXCR4 and CXCR7 downregulation by siRNA on the biological characteristics of endometrial carcinoma cell HEC-1-A. Methods: siRNAs targeting CXCR4 and CXCR7 were transfected into HEC-1-A by using LipofectamineTM2000. The downregulation of CXCR4 and CXCR7 was assessed by RT-PCR and immunoblotting. The effects of altered CXCR4 and CXCR7 expression on cell proliferation,invasion and apoptosis were measured by CCK-8,Transwell and FCM respectively. Results: Both CXCR4 and CXCR7 were highly expressed in HEC-1-A cell lines. After transfected with specific siRNA,the expression of CXCR4 and CXCR7 gene and protein were downregulated markedly. The proliferation and invasion ability of HEC-1-A cells were significantly decreased compared to those transfected with control siRNA,and the cells were arrested in S phase. Conclusion: Knocking down CXCR4 and CXCR7 inhibits the proliferation and invasion ability of human endometrial cancer cell line HEC-1-A,suggesting that they are potential targets for endometrial cancer therapy.