Exosome Derived MiR-22-3p Promote HCC Progression Induced By Ethanol

[Backgrounds]Hepatocellular carcinoma(HCC)is the main primary malignant tumor of the liver.Liver cancer is a major disease that threatens people’s lives in our country.In the malignant transformation of liver cells and the development of liver cancer,the interaction between different factors is the main reason for this problem.Mi RNA regulates many cellular biological processes.In recent years,mi RNAs in exosomes have attracted extensive attention due to their potential role in understanding the molecular pathogenesis of tumors.Exosomes are extracellular vesicles between 30-200 nm and involved in intercellular communication between HCC cells.Exosomes are released into the extracellular matrix and are responsible for carrying biometric information(proteins,lipids,DNA and RNA,especially mi RNAs)to the target cells of surrounding and distant sites and organs,and act as paracrine signals to mediate cell-to-cell communication.The mi RNA in the exosome is protected by the bilateral membrane structure,thus reducing the degradation of mi RNA and promoting cell-to-cell communication.Exosomes could transfer nucleic acids,lipids and proteins.Exosomes take a significant part in the treatment,occurrence and diagnosis of HCC.More and more evidence shows that exosomes participate in cell-to-cell communication by delivering proteins and nucleic acids to receptor cells,thus affecting the biological process of receptor cells.There are many mi RNAs in different kinds of cells,and it has been reported that there are also mi RNAs in the exosome.The occurrence and progess of liver cancer is affected by many factors.Alcohol may be a very important factor.The annual incidence of alcohol-related cirrhosis accounts for about 15-30% of HCC cases.Therefore,we suppose whether alcohol stress can regulate the development of liver cancer cells through exosome.We did a series of experiments to probe into if alcohol-stimulated liver cancer cells can affect the malignant development of untreated liver cancer cells by secreting exosomes.[Methods](1)The chronic alcohol-induced liver cancer cell model was established by chronictreatment of liver cancer cells with 0.2% alcohol for a long time.(2)The exosome with high purity was extracted by ultracentrifugation for follow-up experiments.(3)The expession of miR-22-3p in cells and exosomes was analyzed by q-PCR.(4)The exosome extracted by ultracentrifugation was co-cultured with hepatoma cells to detect the expression level of miR-22-3p in hepatoma cells.(5)After the exosomes were co-cultured with HCC cells,the malignant transformation ability of hepatocellular carcinoma cells were detected by cell biological behavior experiment in vitro.[Results] In this study,exosomes with high purity were extracted.It was found that chronic alcohol stress increased the expression of miR-22-3p in hepatoma cells in vitro,and increased the expression level of miR-22-3p in normal hepatocytes and exosomes of hepatoma cells.We provide exosomes extracted from chronic alcohol-induced hepatoma cells and co-cultured with untreated hepatoma cells.It was found that chronic alcohol stimulation could transfer miR-22-3p through exosomes to hepatoma cells with low expression of miR-22-3p by increasing the expression of miR-22-3p in exosomes.At the same time,we took the co-cultured cells and ordinary untreated liver cancer cells for corresponding cell behavior experiments,and found that the exosomes of liver cancer cells stimulated by alcohol can promote the malignant transformation ability of liver cancer cells.[Conclusion](1)The exosomes with high purity were extracted successfully.(2)Chronic alcohol stress increased the expression of miR-22-3p in hepatoma cells.(3)Chronic alcohol stress increased the expression of miR-22-3p in exosomes of hepatoma cells.(4)Hepatocellular carcinoma cells can absorb exosomes with high expression of miR-22-3p.(5)Alcohol stress can promote the malignant biological behavior of HCC by increasing the expression of exosome miR-22-3p.