3p21·3区域肺癌相关基因RASSF1A的抑癌功能研究  

Analysis of suppressive role of RASSF1A gene at 3p21.3 in lung cancer cell line A549

作　　者：张森[1] 邵康[1] 张翠艳[1] 周芳[1] 王伟[1] 熊美华[1] 赫捷[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤医院胸外科实验室,北京100021

出　　处：《中华医学杂志》2005年第13期908-911,共4页National Medical Journal of China

基　　金：国家"十五"重大科技攻关专项基金资助项目(2002BA711A06);北京市科委科研基金资助项目(H020220020510)

摘　　要：目的观察将野生型RASSF1A通过质粒载体pcDNA3·1导入不表达该基因的非小细胞肺癌(NSCL)细胞系A549后,肺癌细胞生物学特性的变化。方法通过脂质体Lipofectamine2000将质粒载体pcDNA3·1,pcDNA3·1-RASSF1A分别转染入A549细胞系中,通过流式细胞仪检测瞬时转染该基因对细胞周期的影响以及是否诱发凋亡;通过绘制细胞生长曲线,MTT检测,以及集落形成实验来分析稳定转染细胞的生物学特性;同时分析稳定转染RASSF1A对A549细胞裸鼠转移能力的影响。结果瞬时转染RASSF1A使细胞周期阻断在G1/S期(P<0·001);稳定转染该基因的细胞其增殖能力和集落形成能力均显着下降(P<0·01),同时裸鼠转移能力也明显被抑制(P<0·01)。结论野生型RASSF1A基因的外源性表达能显著抑制肿瘤细胞的生长和转移,提示该基因在肺癌的发生发展中发挥重要作用。Objective Loss of heterozygosity (LOH) at 3p21.3 is a common and early event in the development of lung cancer, implying the presence of tumor suppressor genes (TSGs) that may be involved in the pathogenesis of lung cancer.RASSF1A gene, located at this region, containing the Ras-associated domain, has been considered as a candidate tumor suppressor gene because of high frequency of loss of expression and aberrant promoter hypermethylation in most NSCLCs and SCLCs.This study was designed to transfer the expression vector containing RASSF1A to NSCLC cell line A549 to analyze the effect of this gene's exogenous expression.Methods Transfer the pcDNA3.1 and pcDNA3.1-RASSF1A into A549 cells by lipofectamine 2000 respectively, the apoptosis and the changes of cell cycle of transient-transfected cells were analyses by flow cytometry (FCM), cellular proliferation of the stable-transfected was examined by MTT assay, cell growth curve and colony formation assay.Inhibition of lung metastasis in nude mice of RASSF1A exogenous expression was also investigated. Results The progression of the cell cycle was arrested in G1-phase remarkably induced by RASSF1A (P<0.001); the cell viability,growing speed and colony formation ability were decreased obviously by exogenous expression of RASSF1A (P<0.01).The lung metastasis in nude mouse was also significantly inhibited (P<0.01). Conclusion The experimental results show that RASSF1A plays an important roles in origination, progression and metastasis of lung cancer.

关 键 词：3p21.3区域 肺癌相关基因 功能研究 pcDNA3.1 RASSF1A基因 细胞生物学特性 A549细胞系 流式细胞仪检测 抑癌 非小细胞肺癌 细胞生长曲线 稳定转染细胞 集落形成实验 细胞周期阻断 集落形成能力 抑制肿瘤细胞 质粒载体 

分 类 号：R734.2[医药卫生—肿瘤]