反转录病毒介导端粒酶基因对人口腔黏膜角化细胞寿命的影响

发布时间：2018-10-26 10:08

【摘要】：目的:以pBABE-tert重组反转录病毒为载体,介导端粒酶反转录酶基因(hTRT)转染体外培养的人口腔角化细胞,探讨端粒酶对细胞寿命的影响。方法:将含有hTRT的pBABE-tert重组反转录病毒载体扩增后,转染体外培养的人口腔角化细胞,挑选阳性克隆进行传代培养,以PCR-ELISA和PCR-PAGE电泳检测端粒酶活性。结果:获得端粒酶活性稳定表达的口腔角化上皮细胞克隆,寿命可延长至8~9代。结论:转导端粒酶反转录酶基因的人口腔角化细胞寿命延长1倍,但不能永生化,说明人口腔上皮角化细胞的永生化是复杂和多点调控的过程,端粒酶活性增加是细胞永生化的关键但不是唯一原因。
[Abstract]:Aim: to investigate the effect of telomerase on cell life by transfection of telomerase reverse transcriptase gene (hTRT) into cultured human oral keratinocytes using pBABE-tert recombinant retrovirus as a vector. Methods: the recombinant pBABE-tert retrovirus vector containing hTRT was amplified and transfected into human oral keratinocytes in vitro. The positive clones were selected for subculture and telomerase activity was detected by PCR-ELISA and PCR-PAGE electrophoresis. Results: oral keratinized epithelial cells with stable expression of telomerase activity were obtained and their life span was prolonged to 8 ~ 9 passages. Conclusion: human oral keratinocytes transducing telomerase reverse transcriptase gene can prolong the life span of human oral keratinocytes by twice, but can not immortalize, indicating that immortalization of human oral epithelial keratinocytes is a complex and multi-point regulation process. Increased telomerase activity is the key but not the only reason for cell immortality.
【作者单位】：上海交通大学医学院附属第九人民医院·口腔医学院口腔黏膜病科上海市口腔医学重点实验室;上海中医药大学附属曙光医院口腔科;上海交通大学组织工程研究中心上海市组织工程重点实验室;
【分类号】：R781.5

【相似文献】