Biological effects of lentivirus-mediated silencing of minichromosome maintenance protein 7 with shRNA on the liver cancer MHCC-97H cells.

ABSTRACT: OBJECTIVE:This study was to specifically silence the minichromosome maintenance protein 7 (MCM7) expressions with lentivirus-mediated RNA interference technique in liver cancer MHCC-97H cells and its biological consequences were investigated. METHODS:Human MCM7 sequence was used for the design of shRNA targeting MCM7 which was then introduced to lentivirus, followed by transfection into MHCC-97H cells. Real time quantitative PCR and Western blot assay were performed to detect the mRNA and protein expression of MCM7 in these cells. MTT assay was performed to detect cell proliferation, flow cytometry to detect cell cycle and apoptosis, scratch-wound assay to detect cell migration ability, and transwell invasion assay to evaluate the invasion of these cells. RESULTS:We successfully constructed LV-mcm7-RNAi expressing MCM7 shRNA. PCR and Western blot assay showed the mRNA and protein expression of MCM7 reduced significantly when compared with negative control group (LV-NC-RNAi) and blank control group (P<0.05). As compared to blank control group and negative control group, the cell proliferation reduced dramatically (P<0.01), cells were mainly arrested in G0/G1 phase and apoptotic cells increased markedly in LV-mcm7-RNAi group. Moreover, cells transfected with LV-mcm7-RNAi showed significant reductions in the invasion and migration as compared to other groups (P<0.05). CONCLUSION:Lentivirus mediated silencing of MCM7 with shRNA in MHCC-97H cells may inhibit the malignant behaviors of MHCC-97H cells (suppressed proliferation and compromised invasiveness), which is related to the cell cycle arrest and increase in apoptosis.

SUBMITTER: Liu J 

PROVIDER: S-EPMC4538142 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature