关键词：胃肿瘤；细胞凋亡；bcl-2基因；反义寡核苷酸
【摘要 】 目的 探讨反义寡核苷酸(ASODN)对bcl-2基因的表达调控及诱导胃癌细胞凋亡的作用。方法 应用MTT方法比较两条人工合成的ASODN直接作用及其以脂质体(DOTAP)为载体对胃癌细胞的抑制效果。应用流式细胞术和RT-PCR方法检测ASODN对bcl-2蛋白和mRNA表达量的调控。应用相差显微镜、电子显微镜、流式细胞术等方法，观察bcl-2ASODN诱导BGC-823胃癌细胞凋亡的效果。结果 MTT实验显示两条ASODN抑制胃癌细胞增殖的作用呈浓度和时间依赖性，且靶位点于mRNA蛋白编码区(ASODN2)和以DOTAP为载体的ASODN对胃癌细胞的增殖抑制效果为佳。ASODN作用于胃癌细胞，降低bcl-2蛋白和mRNA表达。bcl-2ASODN处理胃癌细胞，观察到肿瘤细胞缩小、凋亡小体出现、染色质浓缩、凋亡峰等凋亡特征性改变。结论 bcl-2ASODN可降低bcl-2 mRNA和蛋白表达，诱导胃癌细胞凋亡。
Apoptosis of human gastric cancercells induced by bcl-2 antisense oligodeoxynucleotides
LU Ping, CHEN Junqing. The FirstAffiliated Hospital, China Medical University, Shenyang 110001
【Abstract 】Objective Tostudy the regulation of bcl-2 gene expression and induction of apoptosis by bcl-2antisense oligodeoxynucleotides (AS-ODN) on human gastric cancer cell line BGC-823 invitro.Methods Two bcl-2 AS-ODNs were synthesized, one covering theinitiation sequence of translation of bcl-2 mRNA (AS-ODN1) and the other covering theprotein coding region (AS-ODN2). BGC-823 cells in logarithmic phase of growth werecultured in the presence of free or liposome (DOTAP)-encapsulated AS-ODN. Cell growth wasassessed by MTT method. The expression of bcl-2 at mRNA and protein levels was examined byRT-PCR and flow cytometry, respectively. Electron microscopy and flow cytometry were usedto demonstrate apoptotic changes in AS-ODN-treated cells.Results Both AS-ODNs inhibited proliferation of BGC-823 cells. The inhibitory activity of AS-ODN2 wasstronger than that of AS-ODN1. AS-ODNs encapsulated in liposome led to more markedinhibition of cell growth than free AS-ODNs. Both AS-ODNs reduced bcl-2 expression ofBGC-823 cells at mRNA and protein levels. Apoptosis of BGC-823 cells were demonstrated bythe appearance of apoptotic bodies, chromatin condensation and pre-G1 peak onflow cytometric analysis.Conclusion Antisense oligodeoxynucleotide ofbcl-2 decreases bcl-2 gene expression and induces apoptosis of human gastric cancer cellsin vitro.
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