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目的探讨在低氧环境下将促血管生成素1(Ang-1)转染到人胃癌细胞BGC-823中,检测其对人胃癌细胞凋亡的影响。方法利用腺病毒作为载体将已构建成功的Ang-1基因的重组腺病毒瞬时转染人胃癌细胞株BGC-823。实验分为对照组(未转染Ang-1的正常胃癌细胞)、低氧组(仅以低氧诱导剂氯化钴进行低氧干预)、转染组(仅以Ang-1转染)和低氧转染组(低氧干预加Ang-1转染)。通过流式细胞术检测转染各组凋亡率的改变,再分别使用半定量RT-PCR和Western blot方法检测上述各组Bcl-2和Bax mRNA和蛋白表达水平。结果与对照组、低氧组和转染组比较,低氧转染组胃癌细胞凋亡率明显下降(P<0.01);Bcl-2 mRNA和蛋白表达水平明显增加,BaxmRNA和蛋白水平明显减少(P<0.05)。结论低氧环境下转染Ang-1能够明显上调人胃癌细胞Bcl-2的表达,下调Bax的表达。这可能是其抑制细胞凋亡的机制之一。
Objective To investigate the effect of angiopoietin-1 (Ang-1) on human gastric cancer cell line BGC-823 in hypoxic environment and its effect on apoptosis of human gastric cancer cell line. Methods Adenovirus was used as a vector to transiently infect the human gastric cancer cell line BGC-823 with the constructed recombinant adenovirus of Ang-1 gene. The experiment was divided into control group (normal gastric cancer cells without Ang-1 transfection), hypoxia group (hypoxia intervention with hypoxia-inducing agent only), transfection group (transfection with Ang-1 only) and Hypoxia transfection group (hypoxia intervention plus Ang-1 transfection). The changes of apoptotic rate in each group were detected by flow cytometry. The mRNA and protein expressions of Bcl-2 and Bax in each group were detected by semi-quantitative RT-PCR and Western blot respectively. Results Compared with the control group, hypoxia group and transfection group, the apoptosis rate of gastric cancer cells in hypoxia group was significantly decreased (P <0.01), the expression of Bcl-2 mRNA and protein was significantly increased, Bax mRNA and protein levels were significantly decreased P <0.05). Conclusion Transfection of Ang-1 under hypoxia can up-regulate the expression of Bcl-2 and down-regulate the expression of Bax in human gastric cancer cells. This may be one of its mechanisms of inhibiting apoptosis.