rL⁃RVG抑制人小细胞肺癌细胞系NCI⁃H446的增殖及迁移

rL⁃RVG inhibits the proliferation and migration of human small cell lung cancer cell line NCI⁃H446

目的探讨重组新城疫病毒rL⁃RVG对人小细胞肺癌细胞系NCI⁃H446增殖、迁移的影响及可能的作用机制。方法将NCI⁃H446细胞随机分为3组,分别加入PBS、新城疫病毒(NDV)和rL⁃RVG,PBS为对照组。用免疫荧光法检测RVG及NDV蛋白在细胞内的表达;用MTT法测定病毒的抗增殖情况;划痕实验及Transwell小室法检测细胞迁移情况;免疫荧光法检测相关迁移蛋白E⁃cadherin和MMP2的表达。结果rL⁃RVG在NCI⁃H446细胞中的感染率高,并能够在其内稳定表达;rL⁃RVG对小细胞肺癌细胞的抑制效果强于NDV。rL⁃RVG组划痕距离显著小于NDV组及对照组(P<0.05);rL⁃RVG组穿过小室的细胞数明显少于对照组(P<0.05)。相较于NDV及对照组,rL⁃RVG组的MMP2蛋白表达下降,而E⁃cadherin蛋白表达上调。结论rL⁃RVG能够抑制NCI⁃H446细胞增殖,其机制可能是通过调节E⁃cadherin和MMP2而影响细胞迁移。

Objective To investigate the effect and potential mechanism of recombinant Newcastle disease virus rL⁃RVG on proliferation and migration of human small cell lung cancer cell line NCI⁃H446.Methods NCI⁃H446 cells were divided into 3 groups:incubated with PBS,Newcastle disease virus(NDV)and rL⁃RVG,respectively.Immunofluorescence assay was used to detect the expression of RVG and NDV protein.MTT assay was used to de⁃termine the anti⁃proliferation of the virus.The cell migration was detected by scratch test and Transwell method.Im⁃muno⁃fluorescence assay was used to detect the expression of E⁃cadherin and MMP2 proteins.Results After infec⁃tion,rL⁃RVG stably expressed in NCI⁃H446 cell with high infection efficiency.The scratch distance in rL⁃RVG group was significantly shorter than that in NDV group and control group(P<0.05).In the Transwell experiment,the number of cells crossing the compartment in the rL⁃RVG group were significantly less than that in the control group(P<0.05).MMP2 protein expression was down⁃regulated in rL⁃RVG group,while E⁃cadherin protein ex⁃pression was up⁃regulated(P<0.05),compared with NDV group and control group.Conclusions rL⁃RVG could inhibit the proliferation of NCI⁃H446 cell with the potential mechanism of effects on cell migration through regulating E⁃cadherin and MMP2.