Effect of Genistein inducing liver cancer MHCC97-L cells apoptosis and activity of caspase-3 and caspase-9 through inhibiting JNK pathway
LIU Shengnan1 QIAN Tiantian1 LANG Huiling1 YU Jiaqi1 XIONG Mengqi1 ZHAO Zhongxin2 MEI Qingbu1 LIU Dan1
1.Basic Medical College, Qiqihar Medical University, Heilongjiang Province, Qiqihar 161006, China;
2.Department of General Surgery, the First Affiliated Hospital of Qiqihar Medical University, Heilongjiang Province, Qiqihar 161041, China
Abstract：Objective To explore the effect of Genistein inducing liver cancer MHCC97-L cells apoptosis and activity of caspase-3 and caspase-9 through inhibiting JNK pathway. Methods The experiment was divided into control group and SP600125 (SP) 10, 20, 30 μmol/L groups. Western blotting was carried out to search the optimum working concentration of SP. The experiment was divided into control, Gen, SP and combined groups. Apoptosis morphology, apoptosis index, activity of caspase-3 and caspase-9 were detected by inverted microscope, fluorescence microscope and spectrophotometry respectively. Results SP obviously blocked the expression of P-JNK protein at a minimum working concentration of 20 μmol/L (P ＜ 0.01). Under inverted microscope, chromatin became darker or condensate in each drug group, and cell changes most obvious in combined group. Early apoptotic cells and late apoptotic cells were observed under fluorescence microscope. Compared with control group, apoptotic index of all drug groups was significantly increased (P ＜ 0.01), among which apoptotic index of combined group was highest and pro-apoptotic effect was stronger than that of Gen group and SP group (P ＜ 0.01). Compared with control group, the activity of caspase-3 and caspase-9 in all drug groups were significantly increased (P ＜ 0.01). Compared with Gen group and SP group, the activity of caspase-3 in combination group was the highest (P ＜ 0.01), and the activity of caspase-3 in SP group was higher than that in Gen group (P ＜ 0.01). The activity of caspase-9 in the combination group was higher than that in the Gen group (P ＜ 0.01), while the activity of caspase-9 in the SP group and Gen group and the combined group and the SP group have no significant differences (P ＞ 0.05). Conclusions These results suggest that inhibition of JNK pathway can promote the apoptosis of Gen induced MHCC97-L cells by activating the caspase-dependent mitochondrial pathway.
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2019, Vol. 16 
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