RÉSUMÉ
Objective To investigate the synergistic effects of tannic acid(TA) and cis-dichlorodiamine platinum(CDDP) on hepatocellular carcinoma HepG2 cells and its activation situation of endoplasmic reticulum stress(ERS) pathway.Methods Human hepatocellular carcinoma HepG2 cells were divided into the control group,TA group,CDDP group and TA+CDDP group,and were cultured in vitro for 24 h.The growth inhibitory effect of medication on HepG2 cells was detected by MTT assay.The pharmacodynamics synergistic effect between the two drugs was analyzed by the drug interaction index,drug dose reduction index and equivalent graphical method.The nucleus changes were observed by DAPI staining.Real time fluorescent quantitative PCR(q-RT-PCR) and Western blot were used to detect the levels of ERS markers glucose regulated protein (GRP)78 and GRP94.Results TA and CDDP had dose-dependent growth inhibition effect on HepG2 cells,their median effective concentrations(IC50) were 360.00 μmol/L and 1.80 μg/mL respectively.The combination treatment of 180.00 μmol/L TA and 0.90 μg/mL CDDP on HepG2 cells could enhance the inhibitory effect on cell growth.Ta and CDDP had synergistic effect for inhibiting hepatocellular carcinoma cells growth.Compared with the TA group and CDDP group,cell shrinkage and rounding were accelerated in the combined group,apoptotic cells were increased,nuclear had pyknosis,irregular edge and dense staining,nuclear fragmentations were increased and the expressions of GRP78 and GRP94 were up-regulated.Conclusion TA can enhance the effect of CDDP on anti-hepatic carcinoma HepG2 cells,and the synergy mechanism may be related to the activation of ERS pathway.
RÉSUMÉ
Objective To detect the activation levels of endoplasmic reticulum stress ATF6-CHOP pathway,and to investigate the molecular mechanism of tannic acid (TA) combined with cisplatin (CDDP) against hepatocellular carcinoma.Methods HepG2 cells were cultured with 180 μM TA or/and 0.9 μg/ml CDDP for 24 h or 48 h.The levels of ATF6 (ATF6α),ATF6B (ATF6β) and CHOP were analyzed by real-time fluorescence quantitative PCR (q-RT-PCR) and western blot.Results We found that after 24 h or 48 h,compared with the control group,ATF6 mRNA and protein levels,ATF6B protein level,CHOP mRNA and protein levels were significantly increased in the TA group,CDDP group or combination group (P ＜ 0.01 or P ＜ 0.05).Conclusion TA can combine with CDDP to increase the levels of endoplasmic reticulum stress ATF6-CHOP pathway,and the ATF6-CHOP pathway may be one of the molecular mechanisms synergistic anti-hepatocellular carcinoma effect of TA and CDDP on HepG2 cells.
RÉSUMÉ
AIM:To investigate the effect of cis-dichlorodiamine platinun ( cDDP) resistance on proliferation , apoptosis, migration and angiogenesis of esophageal cancer cell line KYSE 150.METHODS:Using the method of increa-sing concentration of cDDP in culture for 10 months, the human esophageal carcinoma cDDP-resistant cell line named KYSE150/cDDP was established successfully .The drug sensitivity was measured by MTT assay .The changes of the biolog-ical behaviors between the parental cell line and resistant cell line were determined by morphological observation assay , MTT assay, colony formation assay , DAPI staining, wound healing assay and tube formation experiment .RESULTS: No significant morphological difference between KYSE 150 cells and KYSE150/cDDP cells was observed .Compared with KYSE150 cells, the drug resistance index of KYSE150/cDDP cells was 6.35, and the viability of KYSE150/cDDP cells was decreased.The colony formation rate of KYSE150/cDDP cells was (15.00 ±3.05)%, while the colony formation rate of KYSE150 cells was (86.70 ±6.57)%.The apoptotic rate of KYSE150/cDDP cells was (0.63 ±0.09)%, and that of KYSE150 cells was (8.46 ±1.33)%.Compared with KYSE150 cells, KYSE150/cDDP cells showed a stronger healing ability of scratch, and the migration rate was higher than that of KYSE 150 cells.The results of tube formation experiment showed that the vessel number in KYSE150/cDDP group was 76.20 ±3.18, while the vessel number in KYSE150 group was 50.60 ±1.33.The protein expression of MMP-2 and VEGFR2 in KYSE150/cDDP cells was higher than that in KYSE150 cells.CONCLUSION: KYSE150/cDDP cells present drug-resistant phenotype and show a slow growth rate . The ability of apoptosis is decreased , and the abilities of cell migration and angiogenesis are increased .This may be an im-portant reason for the failure of clinical chemotherapy for esophageal cancer .