摘要
目的 探讨盐酸小檗碱对人胃癌细胞凋亡与P^53，Bcl-2凋亡基因mRNA及蛋白表达水平的影响。方法 以人胃腺癌细胞株SGC-7901为实验模型，用盐酸小檗碱大（8μg／ml）、小（4gg／ml）两个剂量进行干预，运用光镜、扫描电镜（SEM）、琼脂糖凝胶电泳，流式细胞术（FAC）、末端DNA标记法（TUNEL）对凋亡细胞进行定性与定量检测，免疫组化技术、逆转录多聚酶链式反应（RT-PCR）检测细胞中P^53和Bcl-2mRNA及蛋白表达水平。结果 盐酸小檗碱大，小剂量组作用72h，胃癌细胞DNA出现典型梯状改变，流式细胞仪DNA直方图出现典型的亚二倍体峰（凋亡峰），且随干预药物浓度增加，凋亡率增高，G0/G1期细胞百分比上升，S期细胞百分比下降，细胞阻滞于G0／Gl期。小檗碱作用72h，Bcl-2蛋白及mRNA、突变型p^53蛋白表达水平下调，野生型p^53 mRNA表达水平上调。结论 盐酸小檗碱能诱导人胃腺癌细胞凋亡，且随药物浓度增高，凋亡率亦增高，其机制可能与上调野生型P^53基因，下调Bcl-2基因、突变型P^53蛋白表达有关。
Objective To study the effect of berberine on apoptosis and mRNA transcription and protein levels of p^53, Bcl-2 in human gastric cancer cell SGC-7901. Methods The apoptosis of gastric cancer cells was detected by morphology investigation, scanning electron microscope（SEM）, DNA Ladder, flow cytometry and TUNEL. The express of p^53 and Bcl-2 were detected by the method of immunohistochemistry and RT-PCR. Results There were DNA Ladder and apoptotic peaks in DNA histogram. The Apoptosis Index （AI）. the percentage of G0/G1 of the experimental groups was significantly enhanced and the percentage of S, which were associated with dosage. After treated with berberine for 72h, the Bcl-2 mRNA, protein and mtp^53 protein decreased while the wtp^53 mRNA transcription elevated. Conclusion Berberine can induce apoptosis in human gastric cancer cell. The AI is enhanced which is associated with dosage. The mechanism is to reduce the transcription of Bcl-2, mtp^53 and increase the wtp^53.
出处
《中华临床医学杂志》
2006年第1期1-4,共4页
Chinese Journal of Clinical Practical Medicine
基金
本课题为山东省自然科学基金资助项目[Y2001C19]
