| Background:Primary liver cancer is one of the most common cancers in the world.At present,the main cure methods for early liver cancer are surgery and liver transplantation.However,hepatocellular carcinoma is often found in advanced stages,so chemotherapy is the most common treatment for advanced liver cancer.But,drug resistance of current chemotherapeutics is the main problem affecting the efficacy of chemotherapy.Therefore,it is particularly important to explore the mechanism of drug resistance and to find a new therapeutic targets.LRH-1 belongs to the fifth family of nuclear receptors.It has been reported that LRH-1 is expressed in multiple organs of the human body.Our group and other groups have confirmed that LRH-1 affects the occurrence and development of liver cancer through multiple pathways.Therefore,to further explore whether LRH-1 participates in the study of the drug resistance mechanism of oxaliplatin,a common chemotherapy drug in the treatment of liver cancer,is conducive to providing a theoretical basis for the actual clinical drug resistance problem.Objective:Previous studies have shown that LRH-1 plays an important role in the occurrence and development of multiple malignant tumors,and participates in multidrug resistance of multiple cancers.This study analyzed the growth of different cell in oxaliplatin and the expression of genes in two types of hepatocellular carcinoma cells which under-expressing and over-expressing LRH-1,in order to initially explored the mechanism of LRH-1 regulating liver cancer resistance to oxaliplatin.Methods1.Construct low-expressing hepatocellular carcinoma cell lines using TALENs technology and lentivirus-mediated methods to construct over-expressing cells,and select and obtain two stable cell line models,HepG2LRH-1-and Huh7LRH-1+,using PCR or Western Blot methods to verified.2.The expression of LRH-1 and MDR1 in primary liver cancer cell lines under different concentrations of oxaliplatin was detected by qPCR.3.The CCK-8 method was used to detect the proliferative ability of different cell lines at different or same concentrations of oxaliplatin and calculate their IC50.4.Detect the expression of MDR1 in over-expressing cells and original cell lines,low-expressing cells and original expressing cell lines under different oxaliplatin concentrations by qPCR.Results1.PCR and Western Blot results showed that the over-expressing cell line Huh7LRH-1+ and the low-expressing cell line HepG2LRH-1-were successfully constructed.2.The PCR results showed that with the increase of oxaliplatin concentration,LRH-1 and MDR1 in HepG2 and Huh7 cancer cell lines showed an upward tendency,and there was a correlation with the concentration.3.CCK8 results showed that if low expression of LRH-1 in HepG2 cells,their proliferation ability for four consecutive days showed a downward trend under normal culture conditions and oxaliplatin culture;if overexpression of LRH-1 in Huh7 cell lines,under normal medium culture and oxaliplatin-containing medium culture,the original expressing strain had significantly enhanced proliferation ability for four consecutive days.The results of CCK8 showed that the IC50 value of HepG2LRH-1-cell line was significantly lower than that of HepG2 cell;the IC50 value of Hu17LRH-1+ cell was significantly increased compared to Huh7 cell.4.PCR results showed that LRH-1 was low-expressed in HepG2 cells,and the expression level of resistance-related protein MDR1 was decreased.Over-expression of LRH-1 in Huh7,the expression level of resistance-related protein MDR1 was increased.Conclusion:LRH-1 is related to the proliferation and drug resistance of liver cancer.Silencing LRH-1 can inhibit the proliferation of liver cancer cells,and down-regulate the expression of drug-resistant proteins in liver cancer cells.Over-expression of LRH-1 can promotes the proliferation of liver cancer cells and up-regulates the expression of drug-resistant proteins.In summary,LRH-1 participates in and enhances the resistance of liver cancer to oxaliplatin. |
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