论文部分内容阅读
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。
The effect of tumor antigen-associated mitochondrial protein 12 (TAMP12) on the proliferation and apoptosis of tumor cells was analyzed to investigate the biological function of this protein. The human hepatocellular carcinoma cell line (SMMC-7721) transfected with pcDNA3.1-TAMP12 vector was constructed and the SMMC-TAMP12 cell line stably overexpressed by TAMP12 was obtained. The expression of TAMP12 mRNA was detected by Realtime-PCR. Marker protein expression and subcellular localization. The proliferation rate of the cells was detected by MTT assay. The tumor cells were treated with different concentrations of 5-Fluoron-racil (5-FU) to detect the cell growth cycle and apoptosis, and the activity of apoptosis-related enzymes. The results showed that mRNA and protein in transfected cells were stably overexpressed and localized in mitochondria. TAMP12 can promote tumor cell proliferation and inhibit apoptosis, and is related to the activity of caspase 3/8/9.