Effect And Mechanism Of MTA1-CK19 Signaling Pathway On Colon Cancer Metastasis

Objectives To investigate the effect and mechanism of MTA1-CK19 signaling pathway on colon cancer metastasis.Methods HCT116 colon cancer cells were setup as control group and interference group.RNA interference technique combined with transcriptome sequencing(RNA-seq)was used to analyze the difference of gene expression between control group and interference group,and differentially expressed genes were screened.The mass spectrometry data of colorectal cancer tissues in TCGA database were extracted by c Bio Potal data analysis platform to analyze the correlation between MTA1 and CK19 expression in colorectal cancer tissues.164 cases of colorectal cancer tissues from August 2012 to June 2016 in the Department of Pathology,Cancer Hospital of Chinese Academy of Medical Sciences were selected.Immunohistochemistry was used to detect the expression of MTA1 and CK19 in tumor tissues and the relevance of expression between the two in colorectal cancer tissues was analyzed.The HCT116 cell line overexpressing MTA1 was constructed by using lentivirus infection in colon cancer HCT116 cells,named HCT116MTA1.The cell line knocked out of MTA1 was constructed by Crispr gene editing technology,named HCT116 Cr MTA1,and MTA1 and CK19 protein expression level were detected by Western Blot in HCT116MTA1 and HCT116 Cr MTA1 cells.HCT116MTA1 cell line knockdown of CK19 and HCT116CRMTA1 cell line overexpressing CK19 were constructed by using RNA interference technology and plasmid transfection in HCT116MTA1 and HCT116 Cr MTA1 cells,respectively.The effect of MTA1-CK19 signaling pathway on the invasion ability of HCT116 colon cancer cells was detected by transwell assay.Immunofluorescence staining was used to observe the colocalization of MTA1 and differentially expressed gene CK19 in HCT116 colon cancer cells.An immunoprecipitation assay was used to detect the presence of the physical interaction between MTA1 and CK19.Using the c Bio Portal data analysis platform,the genes coexpressed with MTA1 and CK19 were screened,and the obtained gene sets were taken and the intersection of the gene sets was used for functional enrichment analysis.The STRING database platform was used to analyze the interaction of genes enriched in cell adhesion process and to map the interaction network.Results The results of transcriptome sequencing showed that the expression of CK19 was down-regulated in colon cancer HCT116 cells with MTA1 knockdown,and MTA1 and CK19 were positively correlated with each other at m RNA levels and the difference was statistically significant(P<0.05).In the TCGA database,correlation analysis initially verified the positive correlation between MTA1 and CK19 expression in colorectal cancer tissues and the difference was statistically significant(pearson r=0.33,P<0.05).The expression of MTA1 and CK19 in colorectal cancer tissues was significantly higher than that in adjacent tissues.The difference was statistically significant(P<0.01);MTA1 and CK19 were positively correlated in colorectal cancer tissues(r=0.447,P<0.01);in HCT116 colon cancer cells,MTA1 was positively correlated with CK19 expression,overexpression MTA1 up-regulated CK19 expression,MTA1 silenced inhibited CK19 expression;Silencing CK19 in HCT116MTA1 colon cancer cells attenuated the invasive ability of cells(P<0.05);overexpressing CK19 in HCT116 Cr MTA1 colon cancer cells cell invasive ability was enhanced(P<0.05);MTA1 and CK19 co-localized in the cytoplasm of HCT116 colon cancer cells,and there was physical interaction between MTA1 and CK19.MTA1 and CK19 co-expressed genes are enriched in cell adhesion,m RNA splicing,cell metabolism,and protein synthesis.The interaction network analysis showed that GNB2L1,RPL23 A,EEF1D,EEF1 G,PAICS,HSP90AB1,CTTN,RPL34,RPL7 A,RPS2,RPL24 and SPTAN1 may play an important role in the regulation of colon cancer metastasis by MTA1-CK19 signaling pathway.Conclusions In colorectal cancer,MTA1 is positively correlated with CK19 expression;in colon cancer HCT116 cells,MTA1 physically interacts with CK19;MTA1 can promote colon cancer HCT116 cell invasion by up-regulating CK19.Figure 8;Table 2;Reference 88...