BTBD1O对胰岛β细胞增殖的影响和作用机制研究  

The effects and mechanisms of BTBD10 on the proliferation of islet beta cell

作　　者：刘瑜[1] 谷昭艳[1] 苗新宇[1] 龚艳平[1] 肖彧君[1] 李剑[1] 田慧[1] 李春霖[1] 

机构地区：[1]解放军总医院老年内分泌科,北京100853

出　　处：《中华内科杂志》2012年第2期136-139,共4页Chinese Journal of Internal Medicine

基　　金：国家自然科学基金（30873412）;国家青年自然科学基金（30801201）

摘　　要：目的 观察BTBD10基因在大鼠胰岛β细胞株INS-1细胞过表达后对其增生的影响和可能机制.方法 构建重组真核表达质粒pcDNA4.0-BTBD10,利用脂质体转染INS-1细胞,转染后48 h筛选获得稳定表达细胞株,Western blot方法验证BTBD10表达.四唑盐比色法（MTT法）测定INS-1细胞增殖活性.Western blot方法检测蛋白激酶B（PKB,又名Akt）、磷酸化Akt（p-Akt）、哺乳动物的雷帕霉素靶蛋白（mTOR）和磷酸化-mTOR （p-mTOR）的蛋白表达.结果 成功构建BTBD10过表达的稳定INS-1细胞株,BTBD10的高表达可增加INS-1细胞的增殖活性,促进INS-1细胞p-Akt和p-mTOR的表达,而Akt和mTOR蛋白表达无明显变化.结论 BTBD10在INS-1细胞的过表达可通过促进Akt和mTOR的磷酸化水平而激活Akt/mTOR信号通路,使细胞总体蛋白质翻译水平升高,从而促进INS-1细胞的增殖.Objective To explore the role of BTBD10 overexpression in the proliferation of insulinoma cell line INS-1 and its mechanism. Methods The recombined expression plasmid of pcDNA4.0-BTBD10 was constructed by gene cloning technique and was transfected into INS-1 cell by lipofectamine 2000. The stable overexpression BTBD10 of INS-1 cell was selected at 48th hour after transfection.INS-1 cell proliferation activity was measured by MTT method.The expression of BTBD10,protein kinase B（Akt）,phospho-Akt（p-Akt）,mammal target of rapamycin （mTOR） and phospho-mTOR （p-mTOR） were determined by Western blot.Results The stable overexpression BTBD10 of INS-1 cell was successfully constructed. Overproduction of BTBD10 promoted beta cell proliferation. The phosphorylation of Akt and mTOR was increased and the ratio of p-Akt/Akt and p-mTOR/mTOR was enhanced in the INS-1 overexpressed by BTBD10.But the expression of total Akt and mTOR presented no obvious changes. Conclusion The overexpression BTBD10 of INS-1 cell could activate of Akt/mTOR signalling pathway via stimulating phospho-mTOR and Akt,and enhance overall cell protein translation,so as to promote proliferation of INS-1 cell.

关 键 词：胰岛素分泌细胞 细胞增殖 BTBD10 

分 类 号：R587.1[医药卫生—内分泌]