The Primary Research Of KDM5C Function On Human Hepatoma Cells

Primary liver cancer is in the forefront of the mortality of malignant tumors.It has the characteristics of high incidence,uneasy discovery,rapid development and so on.It is a hot topic in modern molecular biology and medicine.KDM5C(JARID1C or SMCX)belongs to the histone demethylation family,is located on the X chromosome.It contains a ARID domain,a Jmj C domain,a JmjN domain and two PHD type zinc finger proteins.The main function of JmjC domain is to specifically remove the H3K4me3 methyl residue.H3K4me3 is an important histone decorating marker and KDM5C further affect the transcriptional activation of other genes via affecting H3K4me3.In this study,the expression of KDM5C in human liver cancer and adjacent tissues was detected by real-time quantitative PCR.The results showed that the expression of KDM5C in the liver cancer tissues was upregulated compared to the adjacent tissues.The results of hematoxylin eosin(HE)staining and immunohistochemical staining showed that the KDM5C gene is expressed in the nucleus and cytoplasm of liver cancer tissues while mainly in the nucleus.In order to study the function of KDM5C in hepatoma cells,CRISPR/Cas9 was used to knock out KDM5C and stable cell line was established.First of all,KDM5C knockout site was designed by CRISPR/Cas9 target site designing website.Then the target sites were connected to p X458 vector,and the vector was transfected to liver cancer cell HepG2.The KDM5C knockout cell line(KDM5C-KO)was established by the finite dilution,and was verified by sequencing and Western blot.Secondly,the effect of KDM5C on the biological behavior of hepatoma cells was studied.The results of MTT proliferation assay and clone formation assay showed that the proliferation and clonal formation of HepG2 cells was inhibited significantly when KDM5C had been knockout.Cell cycle PI staining showed that the proportion of S phase decreases,after the knockout on KDM5C,which showed a tendency of the decrease in cell proliferation.Further studies showed that the apoptosis of hepatoma cells was induced when KDM5C had been knockout.The results of wound healing assay and Transwell assay showed that the trend of cell migration and invasion was significantly inhibited when KDM5C was knockout.In summary,this study analyzed the expression and localization of KDM5C in hepatocellular carcinoma.Then KDM5C was knocked out by CRISPR/Cas9,and stable knockout cell line was established.Knocking out KDM5C significantly inhibits the proliferation,migration,invasion and cloning formation of hepatoma cells.These results indicate that KDM5C plays a role of promoting the occurrence of tumor in hepatoma cells.