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Suppression of mucin 2 enhances the proliferation and invasion of LS174T human colorectal cancer cells
Author(s) -
Bu Xiaodong,
Li Li,
Li Nan,
Tian Xiaoqiang,
Huang Peilin
Publication year - 2011
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20100876
Subject(s) - rna interference , mucin 2 , mucin , transfection , cell culture , cancer research , cell growth , gene silencing , in vitro , small interfering rna , biology , colorectal cancer , microbiology and biotechnology , chemistry , cancer , gene expression , biochemistry , rna , gene , genetics
Altered expression of MUC2 (mucin 2) is related to tumour development in colorectal cancer. Colorectal mucinous carcinomas are positive for MUC2 expression, whereas MUC2 is down‐regulated in non‐mucinous adenocarcinomas. In the present study, we down‐regulated MUC2 expression by RNAi (RNA interference) and investigated the in vitro and in vivo effects on the proliferation and invasion/migration potential of the LS174T human colorectal cancer cells. The LS174T cell line is a goblet‐cell‐like colorectal cancer cell line that continuously produces high levels of MUC2. Inhibition of MUC2 expression in vitro by transfection of LS174T cells with the recombinant plasmid pcDNA6.2‐GW/EmGFP‐miR‐MUC2 led to the production of a stably transfected MUC2‐RNAi LS174T cell line. The proliferation and invasion/migration of MUC2‐RNAi cells in vitro were significantly higher than those in control cells, as assessed by MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2 H ‐tetrazolium bromide], colony formation and transwell assays. Subcutaneous injection of MUC2‐RNAi LS174T cells into nude mice resulted in the development of subcutaneous tumours visible to the naked eye after 1 week. The growth rate of tumours derived from MUC2‐RNAi LS174T cells was greater than that of tumours derived from control cells. Ki67 and matrix metalloproteinase‐9 proteins were detected by immunohistochemistry in the xenografts. The expression levels of these proteins were higher in the MUC2‐RNAi‐derived xenografts than in xenografts derived from control cells. Although the role of MUC2 in colorectal tumorigenesis is not fully understood, these results strongly suggest a relationship between the proliferation and invasion of LS174T cells and the expression of MUC2.