The Research Of Molecular Mechanism For The Role Of Mig In Anoikis In HBV-related HCC

Objective Primary hepatocellular carcinoma(HCC)is one of the common malignancies.It is prone to early metastasis and relapse after surgery,leading to the poor prognosis.HBV infection is the main cause of primary liver cancer.In China,most of HCC patients are associated with HBV infection.The process of metastasis mainly includes detachment from the primary tumor,invasion into surrounding tissues,infiltration into the vasculature and proliferation in the target organs to form metastases.Only a small number of tumor cells that are detached from the primary tumor survive against anoikis and evade immune surveillance.Therefore,the study on the resistance of tumor cells to anoikis may provide a new approach to intervene in the early metastasis of liver cancer.Methods In this study,we collected serum and surgical specimens from patients with hepatitis B-related liver cancer,recorded patient medical records and followed up regularly.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression of serum Mig in normal and hepatitis B-related liver cancer patients.The expression of Mig in hepatoma cell lines with different metastatic potentials was detected;the model of anoikis in vitro was established and the difference in anoikis rate of HCC cell lines with different metastatic potentials under different concentrations of human recombinant cytokine Mig was detected;HCC cell lines were infected with lentivirus to overexpress or underexpress Mig.Under the anoikis model,the effect of Mig on anoikis of different HCC cell lines was detected by flow cytometry.RNA from treated HCC cell lines was extracted and subjected to transcriptome sequencing to explore related signal pathways and perform subsequent verification.Results(1)In this experiment,we reconfirmed the high expression of Mig in the serum of patients with hepatitis B-related liver cancer with a larger clinical sample size.(2)By measuring the level of Mig expression in hepatoma cell lines with different metastatic potentials,we screened Hep3 B cell lines with high expression of Mig and PLC/PRF/5 cell lines with low expression of Mig for subsequent experiments.(3)In the in vitro anoikis model,we found that the apoptosis rate of PLC/PRF/5 cells overexpressing Mig by lentivirus was higher than that of control cells;the apoptosis rate of Hep3 B cells with lower expression of Mig by lentivirus was lower compared with that of control cells.(4)The RNA of Mig-overexpressing PLC/PRF/5 cells and control cells were subjected to transcriptome sequencing.We screened many downstream signaling molecules on the signaling pathway involved in Mig at the transcriptional level.Conclusions Mig was confirmed to be highly expressed in the serum of patients with hepatitis B-related liver cancer by a larger clinical sample size.In the in vitro anoikis model,lentivirus overexpression of Mig can increase the anoikis rate of PLC/PRF/5 cell line;low expression of Mig by lentivirus can reduce the anoikis rate of Hep3 B cells.At the transcriptome sequencing level,many downstream signaling molecules involved in Mig related anoikis were screened.