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目的探讨导致恶性肿瘤出现细胞染色体非整倍数现象的着丝粒蛋白-H(CENP-H)在舌鳞癌中的表达情况,及其对舌鳞癌细胞增殖的影响。方法舌鳞状细胞癌组织及其癌旁舌黏膜组织组成配对子,采用RT-PCR、Western blot分别检测8对配对子的舌鳞癌系TSCCa、Tca8113及原代培养的舌黏膜细胞中CENP-HmRNA水平和蛋白的表达水平;采用免疫组织化学SP法检测同样8对配对子舌黏膜中CENP-H的表达;采用MTT法、克隆形成实验法分析转染CENP-H对TSCCa细胞增殖能力的影响。结果舌鳞状细胞癌中CENP-H在mRNA、蛋白质水平的表达高于舌黏膜上皮,且差异有统计学意义;免疫组化显示CENP-H高表达,且阳性染色定位于肿瘤细胞的细胞核,而在舌黏膜鳞状上皮细胞中未见表达;MTT法、克隆形成试验显示转染CENP-H后舌鳞癌细胞增殖能力明显高于未转染的对照组细胞(P<0.001)。结论舌鳞癌细胞中高表达的CENP-H对舌鳞癌细胞增殖起到促进作用,在舌鳞癌的发生发展中可能扮演重要角色。
Objective To investigate the expression of CENP-H in tongue squamous cell carcinoma and its effect on the proliferation of tongue squamous cell carcinoma. Methods Tumor squamous cell carcinoma and paracancerous lingual mucosa tissues were paired. The expression of CENP-1, Tca8113 in tongue squamous cell carcinoma and tongue squamous cell carcinoma were detected by RT-PCR and Western blot respectively. HmRNA and CENP-H were detected by SP immunohistochemical method. The expression of CENP-H in the mucosa of 8 paired sublingual tongues was detected by immunohistochemical SP method. The effect of transfection of CENP-H on the proliferation of TSCCa cells was analyzed by MTT assay and clone formation assay . Results The expression of CENP-H mRNA and protein in tongue squamous cell carcinoma was higher than tongue epithelium, and the difference was statistically significant. Immunohistochemistry showed that CENP-H was highly expressed, and positive staining was located in the nucleus of tumor cells. But no expression in lingual mucosa squamous epithelial cells. MTT assay and clonogenic assay showed that the proliferation of tongue squamous carcinoma cells was significantly higher than that of untransfected cells (P <0.001) after CENP-H transfection. Conclusion CENP-H highly expressed in tongue squamous cell carcinoma may promote the proliferation of tongue squamous cell carcinoma and may play an important role in the carcinogenesis of tongue squamous cell carcinoma.