Abstract
Previous studies showed that Math1 homologous to human Hath1 can cause mouse goblet cells to differentiate. In this context it is important that the majority of colon cancers have few goblet cells. In the present study, the potential role of Hath1 in colon carcinogenesis was investigated. Sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa, mucosa adjacent colon cancer and colon cancer samples from 48 patients. Hath1 and Muc2 expression in these samples were tested by immunohistochemistry, quantitative real-time reverse transcription -PCR and Western blotting. After the recombinant plasmid, pcDNA3.1(+)-Hath1 had been transfected into HT29 colon cancer cells, three clones were selected randomly to test the levels of Hath1 mRNA, Muc2 mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time reverse transcription-PCR and Western blotting. Moreover, the proliferative ability of HT29 cells introduced with Hath1 was assessed by means of colony formation assay and xenografting. Expression of Hath1, Muc2, cyclin D1 and p27 in the xenograft tumors was also detected by Western blotting. No goblet cells were to be found in colon cancer and levels of Hath1 mRNA and Hath1, Muc2 mRNA and Muc2 were significantly down-regulated. Hath1 could decrease cyclin D1, increase p27 and Muc2 in HT29 cells and inhibit their proliferation. Hath1 may be an anti-oncogene in colon carcinogenesis.
Highlights
Accompanying changes of foods, the morbidity of colon cancer becomes higher and higher in China (Shin et al, 2006)
PcDNA3.1(+)-Hath1 had been transfected into HT29 colon cancer cells, three clones were selected randomly to test the levels of Hath1 mRNA, mucin 2 (Muc2) mRNA, Hath1, Muc2, cyclin D1 and p27 by quantitative real-time reverse transcription-PCR and Western blotting
Sections obtained from colon adenocarcinoma display cells that appeared crowded without goblet-like features (Figure 1C)
Summary
Accompanying changes of foods, the morbidity of colon cancer becomes higher and higher in China (Shin et al, 2006). Targeted deletion of Math resulted mice intestinal secretory cells, including goblet cells, in failure of differentiate (Yang et al, 2001) and Hath can induce Muc expression (van de Wetering et al, 2002). Some studies have shown that Muc was concerned with colon tumors. Studies have shown that cyclin D1 is concerned with tumorigenesis and development in some tumor, so cyclin D1 is considered as a proto-oncogene (Blok et a1., 2000) Another cell cycle regulation factor, p27 can inhibit various kinds of CDK-cyclin complexes so prevents cell from G1 stage into S stage (Bryja et al, 2004). To research the potential role of Hath in colon carcinogenesis, in the present study, we investigated its expression and Muc level in colon cancer. We observed whether introduction of Hath would alter the proliferative ability of colon cancer cells by in vitro and in vivo studies
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