乙酰肝素酶反义寡核苷酸诱导胃癌细胞凋亡的实验研究

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HPA mRNA的表达：并通过下调bcl-2，上调p53的表达，诱导胃癌细胞凋亡。

Experimental study on apoptosis induced by heparanase antisense oligodeixynucleotides in gastric carcinoma cell line SGC-7901

Objective: To study the effect of heparanase(HPA) antisense oligodeoxynucleotides(AS-ODN) on apoptosis in SGC-7901 cell line and its mechanism. Methods: The AS-ODN of HPA-mRNA and its control, scrambled nonsense oligodeoxynucleotide(NS-ODN) were designed and synthesized and phosphorothioated. The ODNs were embedded in cationic liposome lipofectin and transfected into SGC-7901 cells. The semi-quantitative RT-PCR were performed to evaluate the heparanase, bcl-2 and p53 genes. TUNEL staining and FCM were used to detect the effects on cell apoptosis. Results: After transfected, HPA-mRNA and bcl-2 were decreased, p53 was increased. The apoptotic rates and index of SGC-7901 cells were increased. The typical change of cell apoptosis occurred. Conclusion: AS-ODN has a significant inhibitory effect on the expression of HPA-mRNA and cell apoptosis which is related with upregulate of p53 and downregulate of bcl-2 in human gastric carcinoma cell line SGC-7901.