MicroRNA-205-5p Regulates The Chemotherapeutic Resistance Of Hepatocellular Carcinoma Cells By Targeting PTEN/JNK/ANXA3 Pathway

Objective:The expression of microRNA-205-5p(miR-205-5p)in hepatoma carcinoma cells which are treated by 5-fluorouracil(5-Fu)is detected through the real-time quantitative polymerase chain reaction(qRT-PCR).We will define that whether miR-205-5p regulates chemotherapeutic resistance of hepatocellular carcinoma and further explore the mechanism of miR-205-5p.We hope that our study can provide the experimental basis for improving effects of hepatocellular carcinoma chemotherapy and finding new anti-cancer targeted drugs.Methods: The first part,the expression of miR-205-5p in the tissues of hepatocellular carcinoma and peritumor,hepatocellular carcinoma cells and the normal liver cells are detected through the qRT-PCR to define whether miR-205-5p functions as oncomiR or tumor suppressor in the oncogenesis and development of hepatocellular carcinoma.The second part,the IC50 of Bel-7402(Bel)and Bel-7402/5-Fu(Bel/Fu)cells to 5-Fu were determined by CCK-8 kit.The expression of miR-205-5p in the Bel cells which are treated by different concentration of 5-Fu are detected through the qRT-PCR,then the expression of miR-205-5p in Bel and Bel/Fu cells,finally to define that whether miR-205-5p regulates the chemotherapeutic resistance of hepatocellular carcinoma cells.MiR-205-5p mimic and miR-205-5p inhibitor are transiently transfected into Bel and Bel/Fu cells,then the expression of miR-205-5p is detected and CCK-8 kit is used to detect the effect of Bel and Bel/Fu cells’ proliferation after the treatment of 5-Fu.Finally,bioinformatics analysis is performed to detect whether the PTEN is regulated by miR-205-5p.Western blot is used to verify whether miR-205-5p regulates the chemotherapeutic resistance of hepatocellular carcinoma through PTEN/JNK/ ANXA3 pathway.Result:(1)The expression of miR-205-5p in hepatocellular carcinoma tissues and cells are reduced compared with peritumoral tissues and normal liver cells.(2)The IC50 of Bel cells to 5-Fu is 5.498 μg/ml,the IC50 of Bel/Fu cells to 5-Fu is 1073 μg/ml.The expression of miR-205-5p in Bel cells is significantly induced by 5-Fu treatment.The expression level of miR-205-5p in Bel/Fu cells is upregulated compared with Bel cells.The expression of miR-205-5p is significantly upregulated in Bel cells transiently transfected by miR-205-5p mimic,and the expression of miR-205-5p is downregulated in Bel/Fu cells transiently transfected by miR-205-5p inhibitor.Transiently transfecting miR-205-5p mimic reduce the sensitivity of Bel cells to 5-Fu.On the contrary,transiently transfection miR-205-5p inhibitor improve the sensitivity of Bel/Fu cells to 5-Fu.The PTEN is a directed targeted gene of miR-205-5p and miR-205-5p regulates the chemotherapeutic resistance of hepatocellular carcinoma through PTEN/JNK/ ANXA3 pathway.Conclusion:(1)MiR-205-5p functions as a tumor suppression in the hepatocarcinogenesis.(2)The expression of miR-205-5p in Bel cells is significantly induced by 5-Fu treatment,andthe expression level of miR-205-5p in Bel/Fu cells is upregulated compared with Bel cells,which explain that miR-205-5p may regulate the chemotherapeutic resistance of hepatocellular carcinoma.MiR-205-5p inhibitor combined with 5-Fu can improve the sensitivity of hepatocellular carcinoma chemotherapeutic resistancecells to 5-Fu.The PTEN is a directed targeted gene of miR-205-5p.MiR-205-5p regulates the chemotherapeutic resistance of hepatocellular carcinoma through PTEN/JNK/ ANXA3 pathway.All results can provide the experimental basis for improving effects of hepatocellular carcinoma chemotherapy and finding new anti-cancer targeted drugs.