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目的研究壁虎醇提物(GEE)对胶质瘤C6细胞生长的抑制作用并探讨其分子机制。方法设置空白组、对照组(0.01 mg·m L-15-氟尿嘧啶)和实验组(0.1,0.15,0.2,0.3,0.4 mg·m L-1GEE)。噻唑蓝(MTT)法检测GEE对C6细胞增殖的影响;GEE作用于C6细胞24 h后,倒置显微镜观察C6细胞的形态学变化;Hoechst 33258染色法检测C6细胞核凋亡的变化;Western blot检测半胱氨酸天冬氨酸蛋白酶-9(caspase-9)、凋亡诱导因子(AIF)蛋白的表达。结果 GEE作用于C6细胞48 h后,MTT结果显示GEE可抑制C6细胞的增殖,5个剂量实验组的抑制率分别为19.9%,28.7%,63.1%,75.4%,76.3%,与空白组比较差异均有统计学意义(P<0.05)。显微镜下可见典型的凋亡形态学变化。GEE可诱导caspase-9、AIF蛋白表达增加。结论 GEE可抑制C6细胞增殖,可能是通过增加caspase-9、AIF蛋白表达而引起凋亡。
Objective To study the inhibitory effect of gecko alcohol extract (GEE) on the proliferation of glioma C6 cells and to explore its molecular mechanism. Methods The blank group, control group (0.01 mg · m L-15-fluorouracil) and experimental group (0.1,0.15,0.2,0.3,0.4 mg · m L-1GEE) were set up. The effect of GEE on the proliferation of C6 cells was detected by MTT assay. The morphological changes of C6 cells were observed under inverted microscope after GEE treatment for 24 h, the apoptosis of C6 cells was detected by Hoechst 33258 staining, Caspase-9, and apoptosis-inducing factor (AIF) protein expression. Results After GEE treatment for 48 h in C6 cells, MTT assay showed that GEE inhibited the proliferation of C6 cells. The inhibition rates of the 5 doses of GEE were 19.9%, 28.7%, 63.1%, 75.4% and 76.3%, respectively. Compared with the blank group The differences were statistically significant (P <0.05). A typical apoptosis morphological changes can be seen under the microscope. GEE can induce caspase-9, AIF protein expression increased. Conclusion GEE can inhibit the proliferation of C6 cells, which may be caused by increasing the expression of caspase-9 and AIF proteins.