【摘要】 分析高表达肿瘤抗原-线粒体蛋白12(tumor antigen-associated mitochondrial protein 12,TAMP12)对肿瘤细胞增殖和凋亡的影响,探讨该蛋白的生物学功能。构建pcDNA3.1-TAMP12载体转染的人肝癌细胞株(SMMC-7721),获得稳定高表达TAMP12的SMMC-TAMP12细胞株.通过Realtime-PCR检测TAMP12 mRNA表达含量,激光扫描共聚焦显微镜观察双色荧光标记蛋白的表达和亚细胞定位。采用MTT法检测细胞的增殖速率,并应用不同浓度的5-氟尿嘧啶(5-Fluoron-racil,5-FU)处理肿瘤细胞,检测细胞的生长周期和凋亡,以及凋亡相关酶的活性。结果显示转染细胞中mRNA和蛋白呈稳定高表达,定位于细胞线粒体中。TAMP12具有促进肿瘤细胞增殖作用,抑制细胞凋亡作用,并与胱天蛋白酶3/8/9(caspase3/8/9)的活性有关。更多还原

【Abstract】 To observe the effect of the highly expressed tumor antigen-associated mitochondrial protein 12(TAMP12) on the proliferation and apoptosis of tumor cells and to explore the biological function of this protein.pcDNA vector was recombinated and transfected into the human liver cancer cell line(SMMC-7721).The expression of TAMP12 mRNA was detected by real-time PCR and the subcellular localization and quantification of TAMP12 protein labeled with double fluorescein were observed under cofocal laser scanning microscope(CLSM).The capacity of cell proliferation of SMMC-7721-TAMP12 was measured by MTT assay.Meanwhile,the cycle and apoptosis of tumor cell treated with 5-Fluoronracil(5-FU) with different concentrations as well as the activity of apoptosis-associated enzymes were analyzed.The result showed that the expression of TAMP12 mRNA and protein were stable and high in SMMC-7721-TAMP12.TAMP12 located in mitochondria showed effects on promotion of cell proliferation and inhibition of cell-apoptosis in tumor cells,and its anti-apoptosis function was associated with the activity of caspase3/8/9.更多还原