BCSC-1基因异位表达对人小细胞肺癌细胞增殖的抑制效应  

Inhibition of malignant proliferation of human small cell lung cancer cells by ectopic expression of BCSC-1 gene

作　　者：袁芳[1,2] 鞠吉雨[1] 唐媛媛[1] 邸大琳[1] 王丽娜[1] 孙萍[1] 

机构地区：[1]潍坊医学院免疫学教研室,山东潍坊261053 [2]青州市疾病预防控制中心,山东青州262500

出　　处：《中国癌症杂志》2012年第5期336-341,共6页China Oncology

基　　金：山东省自然科学基金(No:2009CM019);山东省教育厅资助项目(No:2007WZ30)

摘　　要：背景与目的:人乳腺癌候选抑制蛋白1(breast cancer suppressor candidate 1,BCSC-1)基因已被证实是一种新型抑癌基因,在多种肿瘤细胞均存在表达缺失的现象。该研究通过将BCSC-1基因转染至人小细胞肺癌细胞株NCI-H446,探讨BCSC-1基因异位表达对NCI-H446细胞增殖的抑制效应。方法:用PCR扩增BCSC-1 cDNA,构建真核重组表达载体pcDNA3.1/v5-HisB-BCSC-1。通过脂质体把pcDNA3.1/v5-HisB-BCSC-1和空质粒pcDNA3.1/v5-HisB转染入野生型NCI-H446细胞。以转染空质粒pcDNA3.1/v5-HisB的NCI-H446细胞为对照组,野生型NCI-H446细胞为空白对照组。采用流式细胞仪检测细胞周期;MTT法检测细胞增殖;免疫组化确认BCSC-1基因和CD44分子在NCI-H446细胞中的表达。结果:成功构建了真核重组表达载体pcDNA3.1/v5-HisB-BCSC-1,制备了BCSC-1基因异位高表达的NCI-H446稳定细胞株。细胞周期分析显示,异位表达BCSC-1的NCI-H446细胞大部分阻滞在G0/G1期,明显高于对照组和空白组(P<0.01)。MTT法检测显示,异位表达BCSC-1的NCI-H446细胞与对照组、空白组相比,生长速度明显减慢(P<0.05)。免疫组化显示异位表达BCSC-1的NCI-H446细胞CD44表达增高。结论:BCSC-1基因的异位表达对NCI-H446细胞的恶性增殖行为有明显的抑制作用,这种抑制作用可能与细胞周期阻滞和黏附分子CD44表达增高有关。Background and purpose： BCSC-1 gene has been proven to be a new tumor suppressor gene. This study explored the effects of ectopic expression of BCSC-1 gene on the proliferation of human small cell lung cancer NCI-H446 cell by transferring BCSC-1 gene. Methods： The BCSC-1 eDNA was amplified and inserted into pcDNA3.1/v5-HisB, pcDNA3.1/v5-HisB-BCSC-1 and pcDNA3.1/v5-HisB were transfected into wild-type NCI-H446 cells with liposome. FACSort flow cytometry analysis was performed to assess the cell cycle by propidium iodide （PI） stained DNA. The proliferation of cell was detected by MTT method. The expressions of BCSC-1 and CD44 were detected by immunohistochemistry. Results： Eukaryotic recombinant expression vector pcDNA3.1/vS-HisB-BCSC- 1 was successfully constructed. A stable cell line expressing BCSC-1 protein was successfully established. Cell cycle analysis showed that the NCI -H446 cells with ectopic expression of BCSC-1 was mostly blocked in the G0/G1 phase （P〈0.01）. MTT assay showed that the growth of NCI-H446 cells with ectopic expression of BCSC-1 was inhibited （P〈0.05） compared with control and blank groups. The expressions of BCSC-1 and CD44 were higher in the NCI-H446 cells with ectopic expression of BCSC-1 compared with control and blank groups. Conclusion： Ectopic expression of BCSC-1 gene exerts profound inhibitory effect on the malignant proliferation of NCI-H446 cell s. This inhibition may be related to the cell cycle arrest and increased expression of CD44.

关 键 词：BCSC-1基因 NCI H446细胞 生长曲线 细胞周期 CD44 

分 类 号：R734.2[医药卫生—肿瘤]