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目的:运用MoMLV载体转导TNFα基因至SHG-44人胶质瘤细胞,检测生长抑制和凋亡。结果:转基因细胞培养上清液中TNFα蛋白分别为(5198.7±3757.4)pg·ml~(-1)和(3217.4±1180.6)pg·ml~(-1)(P＜0.05),其活性达320.0u·ml~(-1),表达特异的mRNA。转基因细胞G_2~M期缩短而G_0~G_1期延长。凋亡率8%~9%。流式细胞检测到亚G_1期峰。电镜中出现染色质固缩成块状和新月小体。DNA电泳现“梯状”条带。结论:转染TNFα基因造成的细胞生长抑制可能与凋亡有关。
OBJECTIVE: To use MoMLV vector to transduce TNFα gene into SHG-44 human glioma cells and detect growth inhibition and apoptosis. Results: The protein levels of TNFα in culture supernatants were (5198.7±3757.4) pg·ml -1 and (3217.4±1180.6) pg·ml -1 respectively (P<0.05), and the activity was 320.0. u·ml~(-1) expresses specific mRNA. The G2~M phase of transgenic cells was shortened and the G_0~G1 phase was prolonged. The rate of apoptosis is 8% to 9%. The sub-G_1 phase peaks were detected by flow cytometry. Electron microscopy showed chromatin condensation and crescent bodies. DNA electrophoresis is now a “step-like” band. Conclusion: The inhibition of cell growth induced by transfection of TNFα gene may be related to apoptosis.