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目的研究survivin基因反义寡核苷酸转染对人肝癌细胞株SMMC-7721细胞周期、细胞凋亡的作用及其可能的作用机制。方法采用脂质体介导survivin基因反义寡核苷酸转染人肝癌SMMC-7721细胞,透射电镜观察细胞超微结构,流式细胞术检测细胞周期及细胞凋亡,RT-PCR方法检测cyclin B1 mRNA的表达。结果survivin反义寡核苷酸转染后细胞内cyclin B1表达由0.36±0.03增加至0.91±0.03,同时G2-M期细胞及凋亡细胞比例分别由5.81%及0.7%明显增加至42.11%及31.35%,同时细胞超微结构呈典型凋亡样改变。结论survivin基因反义寡核苷酸转染细胞后可以通过诱导cyclinB1的表达,导致G2-M期阻滞,从而诱导细胞凋亡的发生。survivin基因反义寡核苷酸转染可以作为治疗肝癌的重要新方法。
Objective To investigate the effect of survivin antisense oligonucleotide transfection on the cell cycle and apoptosis of human hepatocellular carcinoma cell line SMMC-7721 and its possible mechanism. Methods Liposome-mediated antisense oligonucleotide targeting survivin gene was transfected into human hepatocellular carcinoma SMMC-7721 cells, the ultrastructure was observed by transmission electron microscopy, cell cycle and apoptosis were detected by flow cytometry, and cyclin B1 mRNA expression. Results The expression of cyclin B1 increased from 0.36 ± 0.03 to 0.91 ± 0.03 after transfected with survivin antisense oligodeoxynucleotide and the percentage of G2-M phase and apoptotic cells increased from 5.81% and 0.7% to 42.11% and 31.35%, at the same time, the cell ultrastructure showed typical apoptosis-like changes. Conclusion The survivin gene antisense oligodeoxynucleotides can induce the apoptosis of cells by inducing the expression of cyclinB1, resulting in the arrest of G2-M phase. Survivin gene antisense oligonucleotide transfection can be used as an important new method for the treatment of liver cancer.