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目的通过特异性抑制前列腺癌细胞株PC-3中趋化因子受体4(CXCR4)基因的表达,检测RNA干扰片段对PC-3细胞增殖及侵袭转移能力的影响。方法将CXCR4的特异性RNA干扰腺病毒载体转染至前列腺癌PC-3细胞72h后荧光显微镜观察转染情况;采用RT-PCR法检测转染后CXCR4基因mRNA表达情况;Western blot检测转染后CXCR4蛋白表达情况;MTT法检测PC-3细胞增殖情况;Transwell小室侵袭实验检测对PC-3细胞侵袭转移能力。结果 (1)成功将CXCR4-shRNA重组腺病毒载体转染至前列腺癌细胞PC-3;(2)重组腺病毒转染组CXCR4基因mRNA及蛋白均低于阴性对照组和空白组,差异有统计学意义(P﹤0.01),而阴性对照组与空白组之间差异无统计学意义(P﹥0.05);(3)阴性对照组与空白组细胞增殖迅速,两组之间差异无统计学意义(P﹥0.05),实验组经腺病毒载体转染后细胞增殖程度显著减少,与前两组相比差异有统计学意义(P﹤0.05);(4)CXCR4-shRNA腺病毒载体与空白组及对照组相比能显著抑制PC-3细胞的侵袭力(P﹤0.05),抑制率为57.01%,空白组与对照组相比差异无统计学意义(P﹥0.05)。结论重组腺病毒载体CXCR4-shRNA可有效抑制前列腺癌细胞PC-3中CXCR4基因的mRNA和蛋白的表达,并在一定程度上抑制癌细胞增殖及远处侵袭转移,可作为动物实验的理论基础和前期条件。
Objective To detect the effect of RNA interference fragment on the proliferation, invasion and metastasis of PC-3 cells by specifically inhibiting the expression of chemokine receptor 4 (CXCR4) gene in prostate cancer cell line PC-3. Methods Transfection of CXCR4 specific RNA interference adenovirus into PC-3 cells for 72 hours was performed by fluorescence microscopy. The expression of CXCR4 mRNA was detected by RT-PCR. The expression of CXCR4 mRNA was detected by Western blot. CXCR4 protein expression; PC-3 cell proliferation was detected by MTT assay; invasion and metastasis of PC-3 cells were detected by Transwell cell invasion assay. Results (1) CXCR4-shRNA recombinant adenovirus vector was successfully transfected into prostate cancer cells PC-3; (2) The mRNA and protein expression of CXCR4 in recombinant adenovirus transfected group were lower than those in negative control group and blank group (P <0.01), but there was no significant difference between the negative control group and the blank group (P> 0.05). (3) The proliferation of the negative control group and the blank group was rapid, and there was no significant difference between the two groups (P> 0.05). The proliferation of experimental group was significantly decreased after transfection with adenovirus vector (P <0.05). (4) CXCR4-shRNA adenovirus vector and blank group Compared with the control group, the invasiveness of PC-3 cells was significantly inhibited (P <0.05). The inhibition rate was 57.01%. There was no significant difference between the blank group and the control group (P> 0.05). Conclusion The recombinant adenovirus vector CXCR4-shRNA can effectively inhibit the expression of CXCR4 mRNA and protein in PC-3 cells and inhibit the proliferation and distant metastasis of cancer cells to a certain extent, which may serve as the theoretical basis for animal experiments Preconditions.