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目的观察蓝萼甲素对人肝癌细胞HepG2的抑制和诱导细胞凋亡作用。方法不同浓度蓝萼甲素处理HepG2细胞后,MTT检测24h,48h细胞生长抑制率,LDH试剂盒检测细胞LDH释放量,倒置显微镜观察细胞形态,RT-PCR法对凋亡相关基因Bc-l 2,bax和c-myc的mRNA表达进行检测,流式细胞仪检测细胞凋亡率。结果在1.25~20μmol.L-1剂量范围内,蓝萼甲素对HepG2的生长有显著抑制作用,并呈剂量依赖性。蓝萼甲素5,10,20μmol.L-1能够使HepG2培养液中的LDH释放量显著增加,倒置相差显微镜观察,可见细胞明显皱缩。同时蓝萼甲素可使细胞Bc-l2表达减少,Bax表达增加,c-myc表达减少。结论蓝萼甲素可抑制HepG2细胞增殖并诱导其凋亡,其机制可能与调控Bc-l2,Bax,c-myc的mRNA的表达有关。
Objective To observe the inhibitory effect of Copepodon A on HepG2 cells and the induction of apoptosis. Methods HepG2 cells were treated with different concentrations of calpain A for 24 h and 48 h, MTT assay was used to detect the cell growth inhibition rate. LDH kit was used to detect the cell LDH release. The morphological changes of cells were observed with inverted microscope. The apoptosis-related genes Bc-l 2 , bax and c-myc mRNA expression were detected by flow cytometry apoptosis rate. Results In the dosage range of 1.25 ~ 20μmol.L-1, the blue calmodulin had a significant inhibitory effect on the growth of HepG2 in a dose-dependent manner. Calcein 5,10,20 μmol.L-1 can significantly increase the release of LDH in HepG2 culture medium, the inverted phase contrast microscope, we can see the cells were significantly shrinkage. At the same time, blue callactone can decrease the expression of Bcl-2, increase the expression of Bax and decrease the expression of c-myc. Conclusions Colagenin can inhibit the proliferation and induce the apoptosis of HepG2 cells. The mechanism may be related to the regulation of mRNA expression of Bcl-2, Bax and c-myc.