Suppression of mucin 2 enhances the proliferation and invasion of LS174T human colorectal cancer cells

Authors

  • Xiaodong Bu,

    1. Department of Pathology, School of Medicine, Southeast University, Nanjing 210009, Jiangsu Province, Peoples Republic of China
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  • Li Li,

    1. Department of Pathology, Affiliated Nanjing First Hospital, Nanjing Medical University, Nanjing 210006, Jiangsu Province, Peoples Republic of China
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  • Nan Li,

    1. Department of Pathology, School of Medicine, Southeast University, Nanjing 210009, Jiangsu Province, Peoples Republic of China
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  • Xiaoqiang Tian,

    1. Department of Pathology, School of Medicine, Southeast University, Nanjing 210009, Jiangsu Province, Peoples Republic of China
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  • Peilin Huang

    Corresponding author
    1. Department of Pathology, School of Medicine, Southeast University, Nanjing 210009, Jiangsu Province, Peoples Republic of China
      To whom correspondence should be addressed (email hplwpp@yahoo.cn).
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To whom correspondence should be addressed (email hplwpp@yahoo.cn).

Abstract

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-2H-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.

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