体外诱导人口腔上皮异常增生细胞系建立及癌变实验研究

课题采用脂质体转染法将编码有人类端粒酶催化亚基(hTRT)的载体导入人正常上膜上皮细胞，取得超长寿命正常上皮细胞后，用B(a)P诱导建立人口腔粘膜上皮异常增生细胞系，进而是诱导癌变，检测癌变过程中细胞生物学特性指标，研究其动枋变化规律，为体外研究白白斑癌变和寻求防治方法提供细胞模型。并为口腔粘膜组织工程化研究提供正常上皮细胞库。...

The objective is to establish the method of culturing normal human oral keratinocytes and dyshyperplasia oral keratinocytes, transfer human hTRT (telomerase reverse transcriptase) into oral keratinocytes to extent life-span of the cells and study the possibility of transformed oral keratinocytes by B(a)P. The culture technique for growing normal human oral keratinocytes and dyshyperplasia oral keratinocytes have been established. The cells could be maintained in culture up to 4-5 passages, or 30-50 days. Recombinant pEGFP/hTRT plasmid has been constructed and expressed in 293 cells and oral mucosal keratinocytes. Oral mucosal keratinocytes which had been transfected hTRT by Lipofectin or retrovirus could extend one time lifespan than pre-transfected cells. Dysplastic oral keratinocytes (DOK) is the cells between normal cells and carcinoma cells. B(a)P is not good inducing carcinoma reagent for oral mucosal keratinocytes in vitro. This study provide a good cellular model for oral carcinoma study and a good cell source for construction of tissue engineering oral mucosa.