MMS21 Maintains the Stability of Complex MCM2-7 to Promote Hepatocellular Carcinoma Proliferation
10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.2022.0205
- VernacularTitle:MMS21通过维持MCM2-7复合物稳定性促进肝细胞癌增殖活性
- Author:
Wen-jia LI
1
;
Yuan-xin ZHU
1
;
Kai-shun HU
1
Author Information
1. Medical Research Center， Sun Yat-sen Memorial Hospital//Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation， Guangzhou 510080， China
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
MMS21;
proliferation;
CoIP-MS;
MCMs
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2022;43(2):203-211
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the expression level of MMS21 and its functions in hepatocellular carcinoma. MethodsThe GEPIA2 and CPATC databases were used to analyze the mRNA and protein levels of MMS21 in adjacent and carcinoma tissues； the endogenous expression levels of MMS21 were reduced individually by two siRNAs. Meanwhile， flow cytometry was used to detect the distribution ratio of cell cycle of hepatocellular carcinoma cells. BrdU staining and DNA fiber assay were employed to determine cells proliferation rate， and colony assay was used to evaluate the ability of cell proliferation； the MYC tagged MMS21 was cloned and transfected into SK-hep1 to generate MMS21 stably overexpressed cells. Co-IP combined with mass spectrometry were used to identify MMS21-interacting proteome and map interactive pathway； Western Blot （WB） was performed to determine the effect of MMS21 towards cell cycle-related proteins. ResultsCompared with adjacent tissues （ n = 160 ）， MMS21 was significantly higher expressed in cancerous tissues （ n = 369；P < 0.05） and its high expression was associated with poor prognosis （ P < 0.01 ） in hepatocellular carcinoma； Knockdown of endogenous MMS21 by two siRNAs led to the increase of the proportion of cells in G0/G1 phase （P < 0.001； P < 0.01）， and the decrease of the ratio of cells in S （P < 0.01； P < 0.05）， G2/M （P < 0.05； P < 0.05） phase. Moreover， depletion of endogenous MMS21 greatly attenuated the speed of DNA replication （P < 0.000 1； P < 0.000 1） and the rate of cell proliferation （P < 0.01； P < 0.001）. Mass spectrometry analysis identified the total of 641 high-confidence proteins for MMS21， and interactive pathway analysis showed that these binding proteins were highly correlated with cell cycle-related genes. The results of Western Blot further confirmed this conclusion， and depletion of endogenous MMS21 markedly reduced the protein levels of Microchromosome maintenance protein family （MCMs）. ConclusionsMMS21 is highly expressed in hepatocellular carcinoma and positively correlated with the poor prognosis. MMS21 is closely related to the proliferation of Hepatocellular Carcinoma and regulated the stability of cell cycle related proteins and MCM family.
