Hath1在非黏液性结肠腺癌发病中的抑癌基因作用

Anti-cancer effect of Hath1 in pathogenesis of non-mucinous colon adenocarcinoma

目的:探讨Hath1基因在结肠腺癌发病中的潜在作用。方法:从48例非黏液性结肠腺癌患者的标本中取材,石蜡切片染色后观察正常结肠黏膜、癌旁黏膜和结肠腺癌组织中杯状细胞数量的变化;用定量实时逆转录PCR、免疫组织化学和Western blotting分别检测这3种组织中Hath1基因和Muc2基因的mRNA和蛋白表达。将Hath1基因转化至HT29人结肠癌细胞中,然后随机挑选3个转化了Hath1基因的克隆,用定量实时逆转录PCR检测Hath1和Muc2的mRNA表达,用Western blotting检测Hath1、Muc2、cyclin D1和p27蛋白的表达;通过软琼脂克隆形成实验和裸鼠移植实验,观察将Hath1基因转化到HT29细胞后,HT29细胞增殖能力的变化。结果:在非黏液性结肠腺癌组织中未发现杯状细胞,并且Hath1基因和Muc2基因的mRNA和蛋白表达均显著下调(P<0.05);在HT29细胞中,Hath1基因下调cyclin D1蛋白的表达(P<0.05),上调p27(P<0.05)和Muc2(P<0.01)蛋白的表达;过表达Hath1基因的HT29细胞在体外的克隆形成显著受到抑制(P<0.01),移植至裸鼠时移植肿瘤的体积显著缩小(P<0.01)。结论:Hath1基因表达的下调可能与非黏液性结肠腺癌的发生有关,Hath1基因在非黏液性结肠腺癌的发生中可能起着抑癌基因的作用。

Objective To investigate the potential role for Hath1 in pathogenesis of colon adenocarcinoma.Methods The sections of paraffin-embedded tissues were used to investigate the goblet cell population of normal colon mucosa,mucosa adjacent colon cancer and colon cancer tissues obtained from 48 patients with non-mucinous colon adenocarcinoma.The mRNA and protein expressions of hath1 and Muc2 in these samples were tested by quantitative real-time reverse transcription-PCR,immunohistochemistry and Western blotting.The Hath1 gene was transfected into colon cancer cells HT29,then three clones were selected randomly to test the expressions of mRNA and protein of Hath1 and Muc2 by quantitative real-time reverse transcription-PCR and Western blotting.The protein expressions of cyclin D1 and p27 were also tested by Western blotting.Moreover,the proliferative ability of HT29 cells transfected with Hath1 was observed by means of colony formation assay and xenograft.Results There were not any goblet cells to be found in colon cancr.The expressions of mRNA and protein of Hath1 and Muc2 were significantly down-regulated in colon cancer（P〈0.05）.Hath1 could decrease the expression of cyclin D1 protein（P〈0.05）,increase the expressions of p27 protein（P〈0.05） and Muc2 protein（P〈0.01） in HT29 cells,inhibit these cells to form colonies（P〈0.01） and decrease the size of xenografts（P〈0.01）.Conclusion Down-regulation of Hath1 may concern with non-mucinous colon adenocarcinoma,Hath1 may be an anti-oncogene in pathogenesis of non-mucinous colon adenocarcinoma.