冬凌草甲素诱导胶质瘤SHG44细胞凋亡及其分子机制的研究

Mechanism of Oridonin-induced Apoptosis of Glioma SHG44 Cells

目的研究冬凌草甲素诱导胶质瘤SHG44细胞凋亡及其分子机制。方法采用CCK-8比色法绘制生长曲线,冬凌草甲素分别以0、1.25、2.5、5、10、20、40μmol/L观察对胶质瘤SHG44细胞生长活性的影响;Hoehst33258和TUNEL染色法观察细胞形态的变化,流式细胞仪检测细胞凋亡情况;通过Western blotting分析凋亡相关蛋白Caspase-3、cleaved Caspase-3、Bax和Bcl-2在胶质瘤SHG44细胞中蛋白的表达情况。结果冬凌草甲素作用胶质瘤SHG44细胞24 h和48 h后,细胞的增殖受到明显抑制,且24 h和48 h细胞半抑制率(IC50)的浓度分别是7.865和4.74μmol/L;Hoechst33258和TUNEL染色结果发现形态发生明显改变,出现了典型的细胞凋亡变化;Western blotting检测结果显示冬凌草甲素诱导后,下调凋亡抑制蛋白Bcl-2的表达,并激活了凋亡蛋白Caspase-3和Bax的表达。结论冬凌草甲素对胶质瘤SHG44细胞具有抑制增殖及诱导凋亡的作用,同时调控凋亡相关蛋白Caspase-3、Bcl-2和Bax的表达。

Objective To study tile molecular mechanism of oridonin-induced apoptosis of glioma SHG44 cells. Methods A growth curve was plotted using CCK-8 colorimetric method with different concentrations of oridonin （0, 1.25,2.5,5,10,20, and 40 μmol/L）to observe its effect on the growth of SHG44 cells. Hoechst33258 and terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） staining were used to examine the changes in cell morphology and flow cytometry was used to detect cell apoptosis. Western blotting was used to analyze the expression of apoptosis-related proteins （Caspase-3, cleaved Caspase-3, Bax, and Bcl-2）in SHG44 cells. Results SHG44 cell proliferation was significantly suppressed after 24 and 48 h Oridonin treatment, with a half-maximal inhibitory concentration of 7.865 and 4.74 μmol/L, respectively. Hoechst33258 and TUNEL staining showed changes in cell morphology such as shrinkage and nucleus fragmemation and morphogenesis, which are indicative of apoptosis. Western blotting analysis showed that oridonin inhibited the expression of Bel-2 and activated the expression of Caspase-3 and Bax. Conclusion Oridonin can inhibit the proliferation and induce the apoptosis of SHGd4 cells by regulating the expression of apoptosis-relatod proteins.