摘要
目的:研究慢病毒介导的β4整合素(integrinβ4,ITGB4)shRNA对人非小细胞肺癌(non-small cell lung cancer,NSCLC)裸鼠皮下移植瘤生长的抑制作用。方法:采用实时荧光定量-PCR(real-time fluorogenic quantitative-PCR,RFQ-PCR)和蛋白质印迹法检测ITGB4在不同人NSCLC细胞中的表达水平;慢病毒介导ITGB4shRNA抑制NSCLCH460SM细胞中ITGB4的表达,再用RFQ-PCR和蛋白质印迹法检测ITGB4的表达水平,评价基因沉默效率;裸鼠随机分为3组,皮下分别接种H460SM-NS细胞(对照组)、稳定表达ITGB4shRNA的H460SM-68和H460SM-71细胞。每隔1d测量裸鼠体质量和肿瘤大小,比较各组裸鼠肿瘤生长情况。处死裸鼠,剥离肿瘤,测量肿瘤大小和质量。结果:ITGB4在大多数人NSCLC细胞中均有表达,其中大细胞肺癌H460SM和NCI-H460细胞中ITGB4表达量明显高于NCI-H661细胞(P<0.01),H460SM细胞中ITGB4表达水平明显高于亲本NCI-H460细胞(P<0.01);ITGB4shRNA转染组细胞中ITGB4的表达水平明显低于阴性对照组细胞(P<0.01);ITGB4shRNA转染组皮下移植瘤生长速度明显低于阴性对照组(P<0.01)。结论:ITGB4表达可能与人NSCLC细胞的致瘤、侵袭、转移的表型有关;抑制ITGB4的表达,可以抑制人NSCLC细胞H460SM裸鼠皮下移植瘤的生长。
Objective：To investigate the inhibitory effect of gene silencing by lentivirus-mediated delivery of ITGB4 （integrin β4） shRNA on the growth of subcutaneous xenografts of human NSCLC （non-small cell lung carcinoma） H460SM cells in nude mice. Methods：The expression levels of ITGB4 mRNA and protein in different human NSCLC cell lines were detected by RFQ-PCR （real-time fluorogenic quantitative-PCR） and Western blotting, respectively. Lentivirus-mediated delivery of ITGB4 shRNA was used to inhibit the expression of ITGB4 gene in H460SM cells, and the expression of ITGB4 was determined by RFQ-PCR and Western blotting to evaluate the gene-silencing efficiency. The nude mice were randomly inoculated subcutaneously with H460SM cells without infection （H460SM-NS cells） or H460SM-68/H460SM-71 cells stably expressing ITGB4 shRNA. The xenograft tumor volume was estimated from tumor diameter and the mouse weight was measured every other day. The growth of the xenograft tumor was also compared. Finally, the mice were sacrificed, and the subcutaneous xenograft tumor was dissected to measure the tumor size and its weight. Results：Most human NSCLC cell lines expressed ITGB4. The expression levels of ITGB4 were significantly higher in H460SM and NCI-H460 cells than in NCI-H661 cells （P 〈 0.01）. A significantly higher level of ITGB4 expression was observed in H460SM cells as compared with their parental NCI-H460 cells （P 〈 0.01）. A significant down-regulation of ITGB4 expression was also observed in H460SM cells infected with ITGB4 shRNA as compared with H460SM cells infected with negative control shRNA （P 〈 0.01）. The growth of subcutaneous xenografts of H460SM cells by knockdown of ITGB4 in nude mice obviously slowed down （P 〈 0.01）. Conclusion：ITGB4 expression may be associated with tumorigenesis, invasiveness and metastasis of human NSCLC H460SM cells. Inhibition of ITGB4 expression can reduce the growth of subcutaneous xenografts of human NSCLC H460SM cells in nude mice.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第8期578-584,共7页
Tumor
基金
国家自然科学基金资助项目(编号:81060177)
云南省应用基础研究基金资助项目(编号:2009CD181)
中国人事部留学人员科技活动项目择优资助启动基金[编号:人社厅(2010)412号]
云南省卫生科技资助项目(编号:2009NS079)
关键词
癌
非小细胞肺
整合素β4
RNA
小分子干扰
慢病毒感染
小鼠
裸
Carcinoma, non-small cell lung
Integrin beta 4
RNA, small interfering
Lentivirusinfections
Mice, nude
