B-Myb稳定过表达H1299细胞株的构建与分析  

Establishment and Analysis of Lung Cancer Cells H1299 with Stable B-Myb Over-expression

作　　者：蔡伟[1,2] 王义涛[1,2] 朱远远[1,2] 王森[1,2] 朱慧芳[1,2] 雷云龙[1,2] 李轶[1,2] 张莹[1,2] 张春冬[1,2] 卜友泉[1,2] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国细胞生物学学报》2014年第6期732-739,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81171879;81302263);重庆市科委自然科学基金(批准号:cstc2013jcyjA10043)资助的课题~~

摘　　要：B-Myb是Myb家族的成员之一,在细胞周期和癌变过程中具有重要作用。但其在肺癌中的作用及其分子机制仍不清楚。为了研究B-Myb在肺癌中的作用,构建了B-Myb稳定过表达的H1299肺癌细胞株。流式细胞术和MTT检测的结果表明,B-Myb稳定过表达导致G1期细胞减少,S期细胞增加进而促进细胞增殖;克隆形成实验及Transwell的结果表明,B-Myb稳定过表达显著增强H1299细胞的克隆形成、侵袭及迁移能力。定量RT-PCR检测结果表明,B-Myb稳定过表达显著提高了细胞周期基因CCNA1的表达水平;对CD97和MTSS1等细胞运动相关下游基因的表达则无明显影响。该研究成功构建了B-Myb稳定过表达细胞株,发现了B-Myb过表达可促进肺癌细胞的增殖、侵袭迁移及克隆形成能力,为进一步研究奠定了基础。B-Myb is one of members of the Myb family, and it plays critical roles in both cell cycle and cancer development. However, the role and molecular mechanism of B-Myb in lung cancer were still unclear. To investigate the role of B-Myb in lung cancer, H1299 cells with stable B-Myb over-expression were established. FACS and MTT results showed that stable over-expression of B-Myb led to decreased percentage of G1 phase cells, increased percentage of S phase cells and subsequently promoted cell proliferation. Colony formation assay and Transwell assay indicated that stable over-expression of B-Myb significantly enhanced the colony formation, invasion and migration abilities of H1299 cells. Quantitative RT-PCR results also demonstrated that B-Myb over-expression significantly up-regulated the expression of cell cycle related gene CCNA1. There were no significant changes in the expression of cell motility related downstream genes such as CD97 and MTSS1. Taken together, we have successfully established the B-Myb stable over-expressed cells and found that over-expression of B-Myb can promote proliferation, invasion, migration and colony formation abilities of lung cancer cells.

关 键 词：B—Myb 细胞周期 细胞增殖 侵袭 迁移 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]