pVITRO2-VSVM-MBD2抑制肺癌细胞增殖并诱导其凋亡的实验CSCD

目的探讨β防御素-2(MBD2)与水泡口炎病毒基质蛋白双表达质粒对肺癌细胞增殖的影响及其体外对DC细胞的趋化作用。方法MTT法观察各组质粒转染后肺癌LL/2细胞的增殖抑制率,PI荧光染色观察细胞形态变化,Annexin V-PE/7-AAD双染法检测细胞凋亡情况。树突状细胞(DC)趋化实验观察各质粒转染组LL/2细胞上清液中MBD2的活性。结果pVSVM(Lipofectamine 2000转染细胞后转入pVSVM)、pVSVM+pMBD2共转染组和pVSVM-MBD2双表达质粒转染组(M蛋白会在肿瘤细胞内同时表达)细胞形态变化明显,与脂质体转染组(Lipofectamine 2000转染细胞)相比明显抑制了LL/2细胞的增殖。空白对照组(不转染Lipofectamine 2000)和pMBD2转染组(Lipofectamine 2000转染细胞后转入pMBD2)细胞凋亡率较低,而其他转染组细胞凋亡率明显增加,且双表达质粒具有更强的凋亡诱导作用。pMBD2具有良好的DC趋化作用。结论β防御素-2与水泡口炎病毒基质蛋白共表达质粒能显著抑制肺癌细胞LL/2的增殖并诱导其凋亡,且有体外趋化DC细胞活性的能力。

Experiment on pVITRO2-VSVM-MBD2 Inhibiting Proliferation and Inducing Apoptosis of Lung Cancer Cells

Objective To investigate the effect of co-expression plasmid of MBD2 and VSVM on the proliferation of lung cancer cells and its chemotaxis on DC cells in vitro. Methods MTT assay was used to observe the proliferation inhibition rate of lung cancer LL/2 cells after plasmid transfection in each group. Cell morphological changes were observed by PI fluorescence staining. Cell apoptosis was detected by Annexin V-PE/7-AAD double staining. MBD2 activity was observed by DC chemotaxis experiments. Results pVSVM group, pVSVM+pMBD2 co-transfection group and pVSVM-MBD2 double expression plasmid transfection group significantly inhibited the proliferation of LL/2 cells, compared with liposome transfection group; and the cell morphology changed obviously. The cell apoptosis rates of blank control group and pMBD2 transfection group were lower, while those in other transfection groups were significantly increased, and the double expression plasmid had stronger apoptosis induction effect. pMBD2 had a good chemotactic effect on DC. Conclusion The co-expression plasmid of β-defensin-2 and vesicular stomatitis virus matrix protein could significantly inhibit the proliferation and induce the apoptosis of lung cancer cell line LL/2, and has the chemotactic ability of DC cells activity in vitro.