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腺病毒 5型早期区 1 A( Ad5E1 A)基因是新近发现的一个肿瘤抑制基因 .其产物 E1 A蛋白是多功能转录因子 ,它能从正、负 2个途径调控多种细胞基因的转录 ,具有降低体内致瘤性及抗转移等活性 .为了探讨 E1 A基因对代表肺癌癌前病变的永生化人支气管上皮细胞的生长是否具有抑制作用 ,构建了在真核细胞高表达 E1 A基因的重组质粒 p CEP4- E1 A.通过脂质体介导将 E1 A基因转入永生化人支气管上皮细胞第 1 68代 ( MP1 68)中 ,经潮霉素筛选 ,获得稳定表达 E1 A的永生化人支气管上皮细胞 ( MP1 68- E1 A) .结果表明 :E1 A基因的稳定表达抑制了 HER- 2 / neu基因的表达 .转染细胞 ( MP1 68- E1 A)回复扁平形态、恢复细胞生长的接触性抑制 ,细胞群体生长缓慢 (倍增时间是 MP1 68- vect细胞的 1 .41倍 ) ,细胞周期 G1期阻滞并出现凋亡 ,软琼脂集落形成抑制率达73.86% .结果说明 E1 A基因的稳定表达明显抑制了永生化人支气管上皮细胞的生长 .该作用可能与 E1 A抑制 HER- 2 / neu基因的表达及诱导永生化人支气管上皮细胞凋亡有关 .
The adenovirus type 5 early region 1 A (Ad5E1 A) gene is a recently discovered tumor suppressor gene. The product E1 A protein is a multifunctional transcription factor that regulates the transcription of a variety of cellular genes from both positive and negative pathways. It can reduce tumorigenicity and anti-metastatic activity in vivo. To investigate whether the E1A gene has an inhibitory effect on the growth of immortalized human bronchial epithelial cells that represent precancerous lesions of lung cancer, a recombinant protein with high expression of E1A gene in eukaryotic cells was constructed. Plasmid p CEP4- E1 A. Transfection of E1 A gene into immortalized human bronchial epithelial cells at passage 1 68 (MP1 68) by liposome-mediated selection of hygromycin for immortalized humans stably expressing E1 A Bronchial epithelial cells (MP1 68-E1 A). The results showed that stable expression of the E1 A gene inhibited HER-2/neu gene expression. Transfection of cells (MP1 68-E1 A) to restore flat morphology and restore cell growth in contact Sex inhibition, cell population growth was slow (doubling time was 1.41 times that of MP1 68-vect cells), cell cycle G1 arrest and apoptosis, the inhibition rate of soft agar colony formation reached 73.86%. The results show that the E1A gene Stable expression Significantly inhibited immortalized human bronchial epithelial cell growth. This effect may inhibit the induction and expression of E1 A HER- immortalized human bronchial epithelial cell apoptosis 2 / neu gene.