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目的探讨人类重组肝细胞生长因子(rhHGF)在丝裂霉素C(MMC)诱导人胶质瘤U251细胞凋亡中的作用。方法采用倒置相差显微镜和透射电镜观察细胞凋亡变化,MTT法检测细胞生长抑制率,TUNEL法和流式细胞术(FCM)研究U251细胞凋亡及细胞周期变化,分析不同浓度的rhHGF对低剂量MMC诱导U251细胞凋亡的影响。结果MMC组U251细胞出现细胞凋亡特有的形态学特征,其抑制率和凋亡指数(AI)分别为(43.7±3.2)%和(63.2±3.4)%,较不同浓度的rhHGF加MMC组和空白对照组显著增高(P<0.05)。随着rhHGF浓度的增加,细胞生长抑制率和凋亡指数呈递减趋势。FCM检测结果表明,MMC主要诱导G0/G1期细胞凋亡而出现凋亡峰,凋亡率25.86%;10和20μg/LrhHGF能特异性抑制MMC诱导U251细胞凋亡,凋亡率分别为15.14%和11.12%。结论rhHGF能抑制MMC诱导的U251细胞凋亡,可能与胶质瘤的复发、耐药等临床特性有关。
Objective To investigate the role of recombinant human hepatocyte growth factor (rhHGF) in the apoptosis of U251 cells induced by mitomycin C (MMC). Methods The changes of cell apoptosis were observed by inverted phase contrast microscope and transmission electron microscope. The cell growth inhibition rate was detected by MTT assay. The apoptosis and cell cycle of U251 cells were detected by TUNEL and flow cytometry (FCM). The effects of different concentrations of rhHGF on low- Effect of MMC on Apoptosis of U251 Cells. Results The morphological characteristics of U251 cells were observed in MMC group. The rates of inhibition and apoptosis were (43.7 ± 3.2)% and (63.2 ± 3.4)%, respectively. Compared with MMC group and The blank control group was significantly higher (P <0.05). With the increase of rhHGF concentration, cell growth inhibition rate and apoptosis index showed a decreasing trend. The result of FCM showed that MMC mainly induced apoptosis in G0 / G1 phase, and the apoptotic rate was 25.86%. 10 and 20 μg / L rhHGF could specifically inhibit the apoptosis of U251 cells induced by MMC with the apoptosis rates of 15.14% And 11.12%. Conclusion rhHGF can inhibit MMC-induced apoptosis of U251 cells, which may be related to the clinical characteristics of glioma recurrence and drug resistance.