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血管内皮生长因子C过表达或抑制表达对人小细胞肺癌NCI-H446细胞生物学行为的影响
         
Influences of over-expression of VEGF-C and its inhibition on biological characteristics of NCI-H446 human small-cell lung cancer cells

摘    要
目的:探讨血管内皮生长因子C(vascular endothelial growth factorC,VEGF-C)对人小细胞肺癌细胞株NCI-H446生长、体外侵袭和细胞凋亡的影响。方法:构建VEGF-C过表达慢病毒质粒pHRi-VEGF-C和小干扰RNA(small interference RNA,siRNA)慢病毒表达质粒pHRi-siVEGF-C,经慢病毒包装后分别感染NCI-H446细胞。蛋白质印迹法检测VEGF-C的表达水平,MTT方法检测细胞增殖活性,Transwell实验检测细胞体外侵袭能力,FCM检测细胞凋亡比例。结果:与空载体对照组相比,pHRi-VEGF-C质粒感染入NCI-H446细胞后VEGF-C基因表达水平显著升高,而pHRi-siVEGF-C质粒感染后VEGF-C基因表达水平显著降低。pHRi-VEGF-C质粒感染第5天后,NCI-H446细胞增殖(D值=1.481±0.056)明显高于pHRi-siVEGF-C组(D值=0.838±0.035)和空载体对照组(D值=1.146±0.07),差异有统计学意义(P<0.05);pHRi-siVEGF-C组中每个视野侵袭的细胞平均数为39.78±0.77,显著低于pHRi-VEGF-C组的84.87±1.27和空载体对照组的60.82±1.05,差异有统计学意义(P<0.01)。感染了pHRi-VEGF-C质粒后的NCI-H446细胞凋亡数较空载体对照组减少约90%,差异有统计学意义(P<0.01)。结论:VEGF-C过表达可以显著促进NCI-H446细胞增殖,增强其侵袭能力,抑制细胞凋亡;而VEGF-C表达经RNA干扰后可以显著减缓细胞的增殖趋势,降低NCI-H446细胞的侵袭能力。推测VEGF-C可能成为临床治疗小细胞肺癌的基因治疗靶点之一。
标    签   小细胞肺   血管内皮生长因子C   基因表达   RNA干扰   细胞增殖   细胞凋亡   肿瘤侵润   Carcinoma   small-cell lung   Vascular endothelial growth factor C   Gene expression   RNA interference   Cell proliferation   Apoptosis   Neoplasm invasiveness  
 
Abstract
Objective: To investigate the influences of vascular endothelial growth factor C(VEGF-C) on cell proliferation, invasion and apoptosis of NCI-H446 human small-cell lung cancer(SCLC) cells in vitro. Methods: The plasmids for VEGF-C overexpression(pHRi-VEGF-C) and small RNA interference targeting VEGF-C(pHRi-siVEGF-C) were constructed. Then the NCI-H446 cells were infected with the lentiviral packaging plasmids pHRi(an empty vector), pHRi-VEGF-C and pHRi-siVEGF-C, respectively. The expression of VEGF-C was examined by Western blotting; the cell proliferation was detected by MTT method; the invasive ability was detected by Transwell assay; and the apoptosis rate was determined by flow cytometry(FCM). Results: The expression level of VEGF-C protein in the NCI-H446 cells was significantly increased after lentiviral packaging plasmid pHRi-VEGF-C infection while it was significantly decreased after lentiviral packaging plasmid pHRi-siVEGF-C infection. The cell proliferation was significantly increased after the NCI-H446 cells were infected with pHRi-VEGF-C for five days as compared with that of the NCI-H446 cells infected with pHRi-siVEGF-C or pHRi(1.481±0.056, 0.838±0.035 and 1.146±0.07; P< 0.05). The average number of invasive cells in each visual field was significantly reduced in the pHRi-siVEGF-C-infected NCI-H446 cells as compared with that of the pHRi-VEGF-Cor pHRi-infected NCI-H446 cells(39.78±0.77, 84.87±1.27 and 60.82±1.05; P< 0.01). The number of apoptotic cells was also reduced by 90% in the pHRi-VEGF-C-infected NCI-H446 cells as compared with that of the pHRi-infected NCI-H446 cells(P< 0.01). Conclusion: The overexpression of VEGF-C can obviously promote the proliferative and invasive abilities of NCI-H446 cells, and it also can reduce the apoptosis. Meanwhile, these effects can be inhibited by small RNA interference targeting VEGF-C. The results suggest that VEGF-C may become a target of gene treatment for small-cell lung cancer.

中图分类号 R734.2   DOI 10.3781/j.issn.1000-7431.2012.02.001

 
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所属栏目 基础研究

基金项目 上海市自然科学基金资助项目(编号:09ZR142600)

收稿日期 2011/9/9

修改稿日期 2011/11/20

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引用该论文: ZHANG Jie,CHEN Mo,ZHOU Cai-cun,XU Jian-fang,ZHAO Yin-min,SU Bo. Influences of over-expression of VEGF-C and its inhibition on biological characteristics of NCI-H446 human small-cell lung cancer cells[J]. Tumor, 2012, 32(2): 79~84
张洁,陈墨,周彩存,徐建芳,赵印敏,粟波. 血管内皮生长因子C过表达或抑制表达对人小细胞肺癌NCI-H446细胞生物学行为的影响[J]. 肿瘤, 2012, 32(2): 79~84


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