摘要
目的:探讨慢病毒介导α6整合素(integrinα6,ITGA6)shRNA对人非小细胞肺癌(non-small cell lung cancer,NSCLC)H460SM细胞生长和侵袭的影响。方法:实时定量PCR和Western blotting检测ITGA6在9种NSCLC细胞中的表达。慢病毒介导ITGA6 shRNA感染H460SM细胞后,实时定量PCR和Wesern blotting检测H460SM中ITGA6的表达,显微镜下观察H460SM细胞形态的变化,MTS法检测细胞增殖,软琼脂实验检测H460SM细胞锚定非依赖性生长,流式细胞术检测细胞凋亡和细胞周期,迁移和侵袭实验检测H460SM细胞体外迁移和侵袭能力。结果:ITGA6在7种人NSCLC细胞中均有表达,H460SM细胞中ITGA6表达水平明显高于亲本H460细胞[(8.75±0.09)vs(5.78±0.26),P<0.01]。慢病毒介导ITGA6shRNA稳定感染H460SM细胞后的H460SM-75、H460SM-76细胞中,ITGA6表达水平明显低于阴性对照组H460SM-NS细胞[(0.11±0.04)、(0.22±0.04)vs(1.00±0.01),P<0.01]。H460SM-75、H460SM-76细胞增殖活性与H460SM-NS细胞无差异(P>0.05),且凋亡率、细胞增殖指数、G0/G1期细胞的比例也无差异(P>0.05)。H460SM-75、H460SM-76细胞的克隆形成率明显低于H460SM-NS细胞[(18.87±1.47)%、(18.85±1.11)%vs(20.81±1.38)%,P<0.05],迁移率[(43.92±0.41)%、(24.10±0.33)%vs(100.00±0.50)%,P<0.01]和侵袭率[(7.04±2.96)%、(4.68±0.27)%vs(100.00±6.74)%,P<0.01]也明显低于H460SM-NS细胞。结论:抑制ITGA6的表达可以抑制NSCLC细胞锚定非依赖性生长、迁移和侵袭,但不影响NSCLC细胞增殖和凋亡。
Objective： To investigate the effect of lentivirus-mediated integrin α6 （ITGA6） shRNA on growth and invasion of human non-small cell lung cancer （NSCLC） H460SM cells. Methods ： The expression levels of ITGA6 in 9 human NSCLC cell lines were evaluated by quantitative-PCR （ Q-PCR ） and Western blotting. Lentiviral ITGA6 shRNA were transfected into H460SM cells and ITGA6 expression was detected by Q-PCR and Western blotting; the cellular morphology change was observed under a microscope; cell proliferation was detected by MTS assay; anchorage-independent growth was determined by colony formation assay in soft agar; apoptosis and cell cycle were analyzed by flow cytometry; and cell invasion and migration were determined by invasion and migration assay. Results： ITGA6 was expressed in 7 NSCLC cell lines. A significantly higher level of ITGA6 expression was seen in H460SM cells compared with the parental H460 cells （ [8.75 ±0.09] vs [5.78 ±0.26], P 〈0.01 ）. After H460SM cells were stably transfected with lentiviral ITGA6 shRNA （H460SM-75, H460SM-76 cells）, a significant down-regulation of ITGA6 expression was observed in H460SM-75 and H460SM-76 cells compared with negative control H460SM-NS cells （ [ 1.00 ±0.01 ] vs [0.11 ±0. 041, [0.22 ±0. 041, P 〈 0.01 ）. The cell viability of H460SM-75 and H460SM-76 cells had no difference with H460SM-NS cells （ P 〉 0.05 ） , as well as the apoptotic rate, cell proliferative index and proportion of G0/G1 phase cells as compared with the negative control （P 〉 0.05 ）. The colony formation rate of H460SM-75 and H460SM-76 cells was significantly decreased as compared with H460SM-NS cells （[20.81 ±1.381% vs [18.87±1.471%, [18.85±1.111%, P〈0.05）, and the migration rate （[100.00 ＋0.50]% vs [43.92 ＋0.411%, [24.10 ±0.33]%, P 〈0.01） and invasion rate （ [100.00± 6.74]% vs [7.04±2.96]%, [4.68 ±0.27]%, P 〈0.01 ） were also significantly decreased. Conclusion： Knockdown of ITGA6 expression can inhibit anchorage-independent cell growth, migration and invasion of NSCLC cells, but have no effect on cell proliferation and apoptosis.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2012年第4期355-362,共8页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81060177)
云南省应用基础研究基金资助项目(No.2009CD181)
中国人事部留学人员科技活动项目择优资助启动基金资助项目(No.人社厅[2010]412号)
云南省卫生科技基金资助项目(No.2009NS079)~~
关键词
非小细胞肺癌
α6整合素
锚定非依赖生长
侵袭
迁移
non-small cell lung cancer
integrin α6
anchorage independent growth
invasion
migration
