重组hafp和htert双启动子介导mirna26a特异表达的肝癌基因治疗实验研究-recombinant hafp and htert promoters mediate the specific expression of mirna 26a in hepatocellular carcinoma gene therapy.docxVIP

第二军医大学硕士学位论文 - PAGE 4 Constructe the vector to drive miR-26a expression regulated by a hAFP-TERT dual promoter 2. Whether the hAFP-TERT dual promoter was tumor-specific and able to efficiently control miR-26a expression in liver cancer cells. 3. Whether this new plasmid overexpression miR-26a could suppress human cancer cell growth in vitro and in vivo. 4. To find new target gene of miR-26a and explore its role in suppressing human cancer Our data show that miR-26a expression driven by a hAFP-TERT dual promoter was tumor-specific and decreased the viability of tumor cells by regulating ERα, progesterone receptor (PR) and P53 except for cyclin D2 or cyclin E2 in vitro and in vivo. Our data also show that estradiol (E2) promotes the growth of liver cancer cells similar to breast cancer cells partly via the E2-ERα pathway and that miR-26a significantly down regulates ERα and prevents the stimulation of hepatoma cell growth by E2. These data suggest that ERα, which is regulated by miR-26a, is important for liver tumor cell growth. Moreover, hAFP-TERT dual promoter mediated miR-26a expression could specifically exert potential antitumor activity and provide a novel targeting approach for cancer therapy. Key words: hTERT/hAFP promoter; ERα; MicroRNA26a; Hepatocellular carcinoma; Gene therapy  重组 hAFP 和 hTERT 双启动子介导 miRNA26a 特异表达的肝癌基因治疗实验研究 缩 略 词 表 缩写 BSA英文名称 Bovine serum albumin中文名称 牛血清白蛋白 cDNA comlementary deoxyribonucleic acid互补脱氧核糖核酸 CEA carcino-embryonic antigen癌胚抗原 DEPC diethyl pyrocarbonate二乙基焦磷酰胺 DMEM Dulbecco’s modified eagle’s medium达尔伯克改良伊格尔培养基 DMSO dimethyl sulphoxide二甲基亚砜 ECL Enhanced hemagglutinin增强化学发光 ERα estrogen receptor-α雌激素受体 α FBS fetal bovine serum胎牛血清 GFP green fluorescent protein绿色荧光蛋白 hAFP human alpha fetoprotein人甲胎蛋白 HE hematoxylin-eosin苏木素-伊红 HRP horseradish peroxidase辣根过氧化物酶 hTERT human telomerase reverse transcriptase人端粒酶逆转录酶 Mut mutant突变型 PAGE polyarcylamide gel electrophoresis聚