Study On The Apoptosis Induced By JR6-1Through Endoplasmic Reticulum Pathway In Human Hepatocellular Carcinoma BEL-7402Cells

Cancer has become a common disease that does serious harm to human. Traditional radiotherapy and chemotherapy treatment always produce a lot of detrimental side effects, although they can kill the tumor cells. Therefore, the development of an innovative anticancer drug with high selectivity, high specificity and low toxicity has become the current focus.First, we screened the inhibitions of10μg/mL JR6-1、JR6、XLJG1、XLJG2、 XLJG3. XLJG5、XLJG6、XLJG7、XLJG9、XLJG10、XLJG11、XLJG13on human hepatoma cells HepG2、human hepatoma cells BEL-7402、human lung cancer cells A549、 human colon cancer cells HCT-8、breast cancer cells MCF-7、human gastric cancer cells BGC、human cervical cancer cells Hela、human colon cancer cells、human bladder cancer cells in vitro for48h by MTT method. JR6-1was determined as the most activated compound and BEL-7402was the most sensitive tumor cell. Then we further tested the proliferation inhibition effect of JR6-1on BEL-7402cell lines and compared the effect with that of positive drug DDP, and found that the IC50of JR6-1was similar with that of the positive drug group. The IC50of JR6-1was4.29μg/mL and DDP’s was3.03μg/mLWe detected the apoptosis and the cell cycle of JR6-1on BEL-7402cell lines by flow cytometry. The results showed that apoptosis induced by JR6-1on BEL-7402was late apoptosis, and the apoptosis increased accompany with the increase of JR6-1concentration. Compared with control group, it has statistically significantly meaning, P<0.01. When the concentration of JR6-1was1、3、9μg/mL, the apoptosis rate of BEL-7402cell was16.3%、30%、55.5%. The result of cell cycle detected by flow cytometry showed that low concentration JR6-1may arrested BEL-7402cell in G1phase, but middle and high concentration may arrested in S phase.The results of expression levels of Bip/GRP78、CHOP、p-JNK、p-c-Jun、caspase-3what detected by western blot method showed that, BEL-7402cell lines with JR6-1for48h, the Bip/GRP78、CHOP、p-JNK、p-c-Jun、caspase-3all expressed. And the express quantity increased with the increase of JR6-1concentration. It also has statistically significant meaning, P<0.01.It implied that apoptotic mechanism of BEL-7402cell lines induced by JR6-1might be associated with caspase-dependent pathway. The expressed of Bip/GRP78indicated that occurrence of endoplasmic reticulum stress and the possible apoptotic mechanism was the JNK pathway and the CHOP pathway, which were endoplasmic reticulum stress induced apoptotic pathway.