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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 effect of vascular endothelial growth factor C (VEGF-C) on the growth, invasion and apoptosis of human small cell lung cancer cell line NCI-H446. Methods: VEGFR-C lentiviral plasmid pHRi-VEGF-C and small interfering RNA (siRNA) lentiviral plasmid pHRi-siVEGF-C were constructed and infected with NCI-H446 cells respectively. Western blotting was used to detect the expression of VEGF-C, MTT assay was used to detect the cell proliferative activity, Transwell assay was used to detect the in vitro invasion ability, and FCM was used to detect the percentage of apoptosis. Results: Compared with the control group, the expression of VEGF-C gene in NCI-H446 cells was significantly increased after pHRi-VEGF-C plasmid infection, while the expression of VEGF-C gene was significantly decreased after infection with pHRi-siVEGF-C plasmid . The proliferation of NCI-H446 cells was significantly higher than that of pHRi-siVEGF-C group (D value = 0.838 ± 0.035) and empty vector control group (D value = 1.146 ± 0.07), the difference was statistically significant (P <0.05). The average cell invasion per field in pHRi-siVEGF-C group was 39.78 ± 0.77, significantly lower than that in pHRi-VEGF-C group (84.87 ± 1.27 and The empty vector control group 60.82 ± 1.05, the difference was statistically significant (P <0.01). The number of apoptotic NCI-H446 cells infected with pHRi-VEGF-C plasmid was reduced by about 90% compared with the control group, the difference was statistically significant (P <0.01). CONCLUSION: Overexpression of VEGF-C can significantly promote the proliferation of NCI-H446 cells and enhance its invasion and inhibit the apoptosis of cells. VEGF-C expression can significantly slow down the cell proliferation and reduce the invasion of NCI-H446 cells ability. It is speculated that VEGF-C may be one of the targets of gene therapy in the treatment of small cell lung cancer.