摘要
目的人和小鼠正常体细胞端粒酶的活性，存在着种属差异性．肿瘤细胞中端粒酶活性的变化是否亦存在种属差异性尚不清楚．因此，我们在人和小鼠的肿瘤细胞系中，分别转导细胞因子基因ＩＬ２和（或）ＴＮＦα，以观察其对肿瘤细胞增殖活性及端粒酶活性的影响．方法首先将目的基因ＩＬ２和（或）ＴＮＦα构建入ｐＬＸＳＮ或ｐＧＣＥＮ逆转录病毒载体，用脂质体法转染ＰＡ３１７细胞进行病毒包装，再以病毒颗粒分别感染ＭＫＮ２８，ＭＫＮ４５，Ｔｃａ８１１３及Ｈ２２细胞，后者经Ｇ４１８筛选、挑克隆及扩增培养，然后用ＰＣＲ，ＲＴＰＣＲ及Ｓｏｕｔｈｅｒｎｂｌｏｔｉｎｇ鉴定其外源基因的整合及表达，并制作细胞生长曲线，以观察肿瘤细胞增殖活性变化，同时用ＴＲＡＰＰＣＲ法观察转基因肿瘤细胞端粒酶活性变化．结果人ＭＫＮ２８，ＭＫＮ４５及Ｔｃａ８１１３转导ＩＬ２或ＴＮＦα后，肿瘤细胞增殖受到不同程度的抑制；相应地，其端粒酶活性也受到抑制．而小鼠Ｈ２２细胞转导ＩＬ２和（或）ＴＮＦα后，增殖未受抑制，端粒酶活性亦未受抑制．结论人和小鼠肿瘤细胞系中转细胞因子基因ＩＬ２和（或）ＴＮＦα后，端粒酶活性及肿瘤细胞增殖发生不同的变化，提示肿瘤细胞中端粒酶的作用机制，在人和小?
AIM Telomerase activity of somatic cells and its trans forms in vitro may possess diversity between human and murine. To investigate the diversity of the proliferation and the telomerase activity, we have constructed IL2 and/or TNFα into human and murine tumor cell lines. METHODS First, we constructed IL2 and TNFα genes into pLXSN and pGCEN vector. Secondly, the viral particles transfected into PA317 cells to package. Thirdly, the vira infected the tumor cell lines, such as MKN28, MKN45, Tca8113 and H22. Fourthly, the cell clones were selected by G418,and then amplified by cell culture. Finally, we identified the integration and expression of IL2 and/or TNFα genes with the methods of PCR, RT PCR and Southern blot,and made the proliferation curve and determined the telomerase activity. RESULTS The proliferation of MKN28, MKN45 and Tca8113 cell lines transfected with IL2 or TNFα was inhibited, and the telomerase activity was also inhibited. The proliferation of murine tumor cell line H22 transfected with IL2 and/or TNFα, however, was not inhibited, neither was the telomerase activity . CONCLUSION Variations of the proliferation and telomerase activity indicate that diversity exists between human and murine tumor cell lines in terms of telomerase effect on tumor cell proliferation.
出处
《世界华人消化杂志》
CAS
1999年第3期194-196,共3页
World Chinese Journal of Digestology
基金
国家自然科学基金
