| BackgroundHepatocellular carcinoma（liver cancer）is one of common malignant tumor in our country,the former member of a malignant tumor in our country,which cause serious damage to the health of citizens in our country,the morbidity and mortality is still in a rising trend at present,our country at present still for surgery and radiation and chemotherapy treatments,but along with the extensive use of chemotherapy drugs,there is an obvious phenomenon of tolerance,most patients so looking for new colon cancer therapeutic targets is becoming a hot spot of research.The endoplasmic reticulum is an important part of secreting and transmembrane protein biosynthesis,folding,assembly and transport,so eukaryotic cells have specialized mechanisms to ensure that proteins are sufficiently folded and mature to maintain protein balance.However,a large number of physiological and pathological abnormalities can lead to the accumulation of misfolded proteins in ER（ER）,which is known as ER stress.ER stress is closely related to tumor development,invasiveness,and response to cancer treatment.Bortezomib is a cellular osmotic,reversible,and selective proteasome inhibitor for anticancer drugs that disrupt the cell cycle and induce apoptosis.ObjectiveThe effects of Bortezomib on the survival,proliferation and apoptosis of HCC cells were determined,and the effects of inhibition of endoplasmic reticulum stress on the tolerance of Bortezomib cell lines to HepG2 cells treated with Bortezomib were investigated,so as to provide a new molecular target and a combined drug strategy for the clinical treatment of HCC.MethodsDifferent hepatocellular carcinoma cells（HepG2 and HuH7）were cultured in vitro and treated with bortezomib.CCK8 assay was used to detect the effect of bortezomib on the survival rate and proliferation rate of hepatocellular carcinoma cells.Western blotting and kinase activity detection kit were used to analyze the expression level and activity change of apoptosis-related protein caspase-3.To establish a bortezomib cell line resistant to HepG2 cells（HepG2/Bort）.Western blotting was used to detect differences in the expression of endoplasmic reticulum stress related proteins in HepG2 and HepG2/Bort cell lines.Effect of bortezomib combined with er stress inhibitors 4-pba and TUDCA on survival rate of HepG2/Bort cells;Western blotting was used to detect the effect of bortezomib combined with endoplasmic reticulum inhibitor on cleaved caspase-3expression in HepG2/Bort cells.Results1.Different concentrations of bortezomib reduced the survival rate of HCC cells HepG2 and HuH7 in a dose-dependent manner,with statistically significant differences（P<0.05）.2.Bortezomib inhibited the proliferation rate of HCC cells HepG2 and HuH7,and the difference was statistically significant（P<0.05）.3.Bortezomib at different concentrations significantly increased the expression levels of cleaved caspase-3 in HCC cells HepG2 and HuH7,with statistically significant differences（P<0.05）.4.The IC500 of HepG2/Bortezomib cells was significantly higher than that of HepG2cells（P<0.05）.5.Survival rate of HepG2/Bort cells was improved and caspase-3 activity of HepG2/Bort cells was decreased after treatment with different concentrations of bortezomib（P<0.05）.6.The expression of endoplasmic reticulum stress-related proteins in the Bortezomib cell lines of HepG2 cells was enhanced,with statistically significant differences（P<0.05）.7.Bortezomib combined with endoplasmic reticulum inhibitor 4-pba（or TUDCA）reduced the survival rate of HepG2/Bort cells,with statistically significant differences（P<0.05）.8.Bortezomib combined with endoplasmic reticulum inhibitor 4-pba（or TUDCA）enhanced the expression of cleaved caspase-3 in HepG2/Bort cells,with statistically significant differences（P<0.05）.ConclusionBortezomib can decrease the survival rate,proliferation rate and apoptosis of hepatocellular carcinoma cells.Combination of ER stress inhibitors can improve the sensitivity of HepG2/Bort to bortezomib. |
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