Liu Yu, Tan Jinjing, Li Lin, Li Shuo, Zou Shuangmei, Zhang Ying, Zhang Xiaojing, Ling Bing, Han Naijun, Guo Suping, Gao Yanning
Department of Etiology and Carcinogenesis, Chinese Academy of Medical Sciences, Beijing 100021, China.
Zhongguo Fei Ai Za Zhi. 2010 Oct;13(10):923-7. doi: 10.3779/j.issn.1009-3419.2010.10.12.
The imprinted gene dlk1 has been recognized as a cancer related gene since it aberrantly expressed in a series of cancer tissues, but its role in lung cancer is still unknown. The aim of this study is to examine dlk1's expression in non-small cell lung cancers (NSCLCs) and investigate the molecular mechanism by which dlk1 could accelerate the proliferation of the cells in lung cancer cell lines (H520).
The relative expression of dlk1 among 30 NSCLC specimens and their adjacent normal lung tissues were analyzed by RT-PCR. A cell model that stably expressed exogenous dlk1 was established following that the dlk1 gene was cloned into a eukaryotic expression vector and then transfected into the lung cancer cells H520. CCK8 analysis and colony forming assay were employed to investigate the effect of dlk1 on cell proliferation. The expression of CyclinB1 was detected by Western blot.
dlk1 aberrantly expressed in 36.7% (11/30) of the tumor tissues of NSCLC compared with their adjacent cancer lung tissues. CCK8 analysis showed that overexpression of dlk1 could promote the proliferation of H520 cells (P < 0.05) and the results was further confirmed by colony forming assay. Western blot analysis found that over expression of dlk1 could up-regulate the expression of CyclinB1 (P < 0.05).
dlk1 aberrantly expressed in NSCLCs. The Overexpression of dlk1 could accelerate the proliferation of lung cancer cells H520 in vitro, probably through up-regulating the expression of cell cycle protein CyclinB1.
印记基因dlk1自其在一系列癌组织中异常表达以来,已被公认为与癌症相关的基因,但其在肺癌中的作用仍不清楚。本研究旨在检测dlk1在非小细胞肺癌(NSCLC)中的表达,并探讨dlk1促进肺癌细胞系(H520)细胞增殖的分子机制。
采用RT-PCR分析30例NSCLC标本及其癌旁正常肺组织中dlk1的相对表达。将dlk1基因克隆到真核表达载体中,然后转染到肺癌细胞H520中,建立稳定表达外源性dlk1的细胞模型。采用CCK8分析和集落形成试验研究dlk1对细胞增殖的影响。通过蛋白质免疫印迹法检测细胞周期蛋白B1(CyclinB1)的表达。
与癌旁肺组织相比,36.7%(11/30)的NSCLC肿瘤组织中dlk1异常表达。CCK8分析表明,dlk1过表达可促进H520细胞增殖(P<0.05),集落形成试验进一步证实了该结果。蛋白质免疫印迹分析发现,dlk1过表达可上调CyclinB1的表达(P<0.05)。
dlk1在NSCLC中异常表达。dlk1过表达可能通过上调细胞周期蛋白CyclinB1的表达,在体外加速肺癌细胞H520的增殖。