To Explore The Mechanisms Of MCM7 Affect Human Hepatoma Cell Cycle

Objective:To construct the models aboult silencing MCM7 in human hepatoma SMMC-7721, MHCC-97H, HepG2 cell lines by RNA interference lentiviral vector.Approach:By building capable of silencing MCM7 gene lentiviral RNA interference vector transfected human hepatoma SMMC-7721, MHCC-97H, HepG2 cell lines, thereby silencing the MCM7 gene expression. And verify the efficiency by quantitative real-time PCR and Western blotting.Results:The stable MCM7 gene silencing hepatoma SMMC-7721, MHCC-97H, HepG2 cells were constructed, the MCM7 gene expression at the mRNA and protein levels were reduced.Conclusion:This study utilized a lentiviral vector and RNA interference technology to build the RNA interference lentivirus particles transfected human hepatoma cells, after screening, can successfully build a stable MCM7 gene silencing human hepatoma cell lines.Objective:To explore the mechanisms of MCM7 for regulating the growth of liver cancer cells.Method:Human genomewide expression profile chip was employed to screen the differentially expressed genes in SMMC-7721 cells after silencing MCM7 gene expression, and bioinformatics analysis of the differently expressed genes was performed. Then using the real-time quantitative PCR to verify the expression of the cell cycle-related genes in the SMMC-7721, MHCC-97H, HepG2 cells after silencing MCM7.Results:In total there were 1010 genes that differentially expressed in SMMC-7721 cells after the expression of gene MCM7 was silenced, including 391 up-regulated genes and 619 down-regulated genes. Bioinformatics analysis showed that these differentially expressed genes were involved in many cellular biological processes such as macromolecules metabolism, cell cycle regulation, cell proliferation regulation, apoptosis, endocytosis, P53 and mTOR signaling pathways, etc. The down-regulations of MCM7、SKP2、CCND1、TP53、 MAD2L1、CDC45、CDK6、DBF4 were confirmed by qRT-PCR in the SMMC-7721, MHCC-97H, HwpG2 liver cells after silencing MCM7, the CDC25A were down-regulated in SMMC-7721, HwpG2 cells after silicing MCM7,but no change in MHCC-97H cells; TTK gene were upregulated in SMMC-7721, MHCC-97H cells after silicing MCM7, but upregulated in HwpG2 cell after silencing MCM7. Bioinformatics analysis predicted MCM7 genes by human ubiquitin promoter factor UBB thereby regulating the expression of SKP2 and other cell cycle related genes, and by real-time quantitative PCR confirmed that the gene were downregulated in SMMC-7721, MHCC-97H, HepG2 cells after the silence of MCM7.Conclusions:The differentially expressed genes after silencing the gene MCM7 in liver cancer cells SMMC-7721 might provide some clues for understanding the mechanism by which MCM7 affects the growth of liver cancer cells.