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。
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-C-or 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.
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第2期79-84,共6页
Tumor
基金
上海市自然科学基金资助项目(编号:09ZR142600)
关键词
癌
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
