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目的:探讨乙酰肝素酶(HPA)反义寡脱氧核苷酸(AS-ODN)对人胃癌细胞株SGC-7901细胞凋亡的影响及其作用机制。方法:设计合成HPAAS-ODN,用脂质体介导法转染SGC-7901细胞,用逆转录聚合酶链反应(RT-PCR)检测转染前后细胞HPA-mRNA及p53、bcl-2基因的表达;用倒置相差显微镜及扫描电镜观察转染前后细胞的形态学变化;用DNA末端原位标记染色法(TUNEL)检测转染前后细胞的凋亡指数;用流式细胞仪检测转染前后细胞凋亡率的变化。结果:HPAAS-ODN转染后,SGC-7901细胞内HPA-mRNA表达下降,p53的表达量升高,bcl-2的表达量降低;细胞凋亡指数和凋亡率升高;出现了典型的凋亡细胞的形态学变化。结论:HPAAS-ODN可有效地抑制胃癌细胞HPAmRNA的表达;并通过下调bcl-2,上调p53的表达,诱导胃癌细胞凋亡。
Objective: To investigate the effect of heparanase (HPA) antisense oligodeoxynucleotide (AS-ODN) on the apoptosis of human gastric cancer cell line SGC-7901 and its mechanism. Methods: HPAAS-ODN was designed and synthesized, and transfected into SGC-7901 cells by liposome-mediated method. The expression of HPA-mRNA, p53 and bcl-2 genes were detected by reverse transcriptase-polymerase chain reaction The morphological changes of the cells before and after transfection were observed by inverted phase contrast microscope and scanning electron microscope. The apoptosis index of the cells before and after transfection was detected by TUNEL staining. The cells before and after transfection were detected by flow cytometry Apoptosis rate changes. Results: After HPAAS-ODN transfection, the expression of HPA-mRNA in SGC-7901 cells decreased, the expression of p53 increased and the expression of bcl-2 decreased. The apoptotic index and apoptosis rate of SGC- Morphological changes of apoptotic cells. Conclusion: HPAAS-ODN can effectively inhibit the expression of HPA mRNA in gastric cancer cells, and induce the apoptosis of gastric cancer cells by down-regulating bcl-2 and up-regulating the expression of p53.