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目的运用外源性RNA干扰技术,敲低MDA-MB-231乳腺癌细胞中软骨同源蛋白1(CART1)基因的表达,研究CART1基因沉默后对MDA-MB-231细胞生物学行为的影响。方法采用人工合成的针对CART1基因的siRNA,用转染试剂RNAi MAX将CART1-siRNA转染MDA-MB-231细胞。实时定量PCR检测转染后CART1 mRNA水平,Western blot法检测转染后CART1蛋白水平,TranswellTM侵袭实验检测MDA-MB-231细胞侵袭、CCK-8法检测细胞增殖、流式细胞术检测细胞周期的变化。结果 CART1-siRNA能有效沉默CART1在MDA-MB-231细胞中的表达,CART1沉默后MDA-MB-231细胞侵袭和增殖能力明显下降,S期细胞的比率增高而G2/M细胞比率减少。结论下调MDA-MB-231细胞CART1的表达,可降低其侵袭和增殖能力、诱导S期阻滞。
Objective To investigate the effect of CART1 gene silencing on the biological behavior of MDA-MB-231 cells by knocking down the expression of CART1 gene in MDA-MB-231 breast cancer cells using exogenous RNA interference technology. Methods CART1 siRNA was transfected into human breast cancer cell line MDA-MB-231 by RNAi MAX, a synthetic siRNA against CART1 gene. Real-time quantitative PCR was used to detect the level of CART1 mRNA after transfection. Western blot was used to detect the level of CART1 protein. TranswellTM invasion assay was used to detect the invasion of MDA-MB-231 cells. Cell proliferation was detected by CCK-8 assay. Cell cycle Variety. Results CART1-siRNA could effectively silence the expression of CART1 in MDA-MB-231 cells. The silencing of CART1 significantly decreased the invasion and proliferation of MDA-MB-231 cells, but increased the percentage of S phase cells and decreased the ratio of G2 / M cells. Conclusion The down-regulation of CART1 expression in MDA-MB-231 cells can reduce the invasion and proliferation and induce S phase arrest.