TNF-α重组慢病毒对胃癌细胞自分泌杀伤作用的实验研究

摘要

目的：探讨TNF-α重组慢病毒对胃癌细胞自分泌杀伤作用。方法 TNF-α重组慢病毒感染SGC-7901胃癌细胞为实验组，并设立阴性对照组以及空白对照组，采用RT-PCR检测3组细胞中TNF-αmRNA的表达情况，ELISA检测3组细胞上清液中TNF-α蛋白含量，流式细胞术检测3组细胞凋亡情况。结果 PCR实验观察到TNF-αmRNA荧光条带24h后稳定表达。测得三组培养液中TNF-α含量，实验组[(2.6926±0.7563) ng/ml]明显高于阴性对照组[(0.9326±0.3091) ng/ml]和空白对照组[(0.8552±0.2768) ng/ml]，且差异均有统计学意义（P均0.05）。在流式细胞实验中，实验组细胞凋亡率明显高于其他组，差异有统计学意义（P0.05）。结论 TNF-α重组慢病毒可以感染SGC-7901胃癌细胞，并在胃癌细胞中稳定表达并自分泌TNF-α，诱导杀伤胃癌细胞。%Abstact:Objective To explore the expression of TNF-α gene by Lentiviral vector-mediated in SGC-7901 gastric cancer cells and the apoptosis induced by TNF-α that autocrined in gastric cancer cell.Methods The SGC-7901 gastric cancer cells were cultured. The gastric cancer cells were devided into three groups:1. the experimental group, the gastric cancer cells were infected with lentiviral vector-mediated TNF-αgene in cells, 2,the negative control group ,the gastric cancer cells infected with lentiviral vector cell group and 3,the control group ,SGC-7901 gastric cancer cells only. The expression of TNF-α was observed by PCR and Elisa method. The apoptosis of the gastric cancer cells were observed by flow cytometry.Results The expression of green fluorescent protein(GFP) was found in SGC-7901 gastric cancer cells infected with lentivirus 48h, and the expression efficiency was more than 90% when stable expression of the fluorescent trap on the TNF-α mRNA in experimental group after 24 hour. The TNF-α in the cultured liquid in the experimental group(2.6926±0.7563 ng/ml) was higher than that of in the negative control group (0.9326±0.3091 ng/ml) and in control group (0.8552±0.2768 ng/ml).T he differences were signiifcant (P0.05).Conclusion TNF-α genes mediated by lentivirus successfully infected gastric cancer cells and expressed TNF-α .The trans-TNF-α genes gastric cancer cells could autocrine TNF-α to induce apoptosis.