PTEN基因联合多西环素对黏液表皮样癌细胞增殖的影响

Effects of Combination of PTEN Gene with Doxycycline on Cell Proliferation of Human Mucoepidermoid Carcinoma Cell Line

目的:探讨PTEN抑癌基因联合多西环素(Dox)对黏液表皮样癌细胞系增殖的影响。方法:用脂质体将野生型PTEN基因导入人黏液表皮样癌细胞系,再用不同浓度的多西环素处理细胞,采用MTT比色法测定细胞存活,应用HE染色判定细胞形态,用流式细胞仪检测细胞周期,用免疫组化染色检测细胞PCNA和EGFR蛋白表达。结果:PTEN基因转染联合Dox组癌细胞增殖抑制显著,细胞出现空泡变性和凋亡,多数癌细胞阻滞于G1期,癌细胞中PCNA和EGFR蛋白的表达显著减弱,比PTEN基因转染或Dox单用作用更强。结论:PTEN抑癌基因联合多西环素对黏液表皮样癌细胞系增殖具有显著的协同抑制效应。

Objective： To investigate the role of combination of exogenous wild type PTEN gene with doxycycline on growth of mucoepidermoid carcinoma cell line. Methods： The wild--type PTEN tumor suppressor gene or empty vector was introduced into mucoepidermoid carcinoma cell line in vitro, then the cancer cells were treated with doxy cycline. Cell survival was determined by MTT assay. Cell morphology was evaluated using hematoxylin and eosin staining. Cell cycle was determinded by flow cytometry. The expressions of PCNA and EGFR protein in cells were analyzed by immunohistochemical methods. Results： In the group of combination of PTEN gene with doxycycline, cancer cell growth was remarkably inhibited, and typical vacuole denaturalization and apoptosis in cells were ob served. The majority of cancer cells were arrested in G1 phase and the level of PCNA and EGFR protein expression in cells was markedly decreased. The combination of PTEN gene with doxycyeline demonstrated more significant in hibitory effect than PTEN gene transfection or doxycycline alone. Conclusion： PTEN gene in combination with dox- ycycline has significant synergistic inhibitory effect on cell proliferation of mucoepidermoid carcinoma.