【摘要】：目的:以pBABE-tert重組反轉(zhuǎn)錄病毒為載體,介導(dǎo)端粒酶反轉(zhuǎn)錄酶基因(hTRT)轉(zhuǎn)染體外培養(yǎng)的人口腔角化細(xì)胞,探討端粒酶對(duì)細(xì)胞壽命的影響。方法:將含有hTRT的pBABE-tert重組反轉(zhuǎn)錄病毒載體擴(kuò)增后,轉(zhuǎn)染體外培養(yǎng)的人口腔角化細(xì)胞,挑選陽(yáng)性克隆進(jìn)行傳代培養(yǎng),以PCR-ELISA和PCR-PAGE電泳檢測(cè)端粒酶活性。結(jié)果:獲得端粒酶活性穩(wěn)定表達(dá)的口腔角化上皮細(xì)胞克隆,壽命可延長(zhǎng)至8~9代。結(jié)論:轉(zhuǎn)導(dǎo)端粒酶反轉(zhuǎn)錄酶基因的人口腔角化細(xì)胞壽命延長(zhǎng)1倍,但不能永生化,說(shuō)明人口腔上皮角化細(xì)胞的永生化是復(fù)雜和多點(diǎn)調(diào)控的過(guò)程,端粒酶活性增加是細(xì)胞永生化的關(guān)鍵但不是唯一原因。
[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.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬第九人民醫(yī)院·口腔醫(yī)學(xué)院口腔黏膜病科 上海市口腔醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室;上海中醫(yī)藥大學(xué)附屬曙光醫(yī)院口腔科;上海交通大學(xué)組織工程研究中心上海市組織工程重點(diǎn)實(shí)驗(yàn)室;
【分類號(hào)】：R781.5

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