Yan Yulan, Liang Bing, Zhang Jin, Jia Lijuan, Liu Yang, Zhang Jie
Department of Respiratory Medicine, Affiliated People's Hospital, Jiangsu University, Zhenjiang 212002, China.
Department of Respiratory Medicine, Affiliated People's Hospital, Jiangsu University, Zhenjiang 212002; Clinical Medicine College, Jiangsu University, Zhenjiang 212001, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 May;30(5):449-52, 457.
To infect lung adenocarcinoma A549 cells with recombinant LaSota strain Newcastle disease virus (NDV) vaccine expressing the rabies virus glycoprotein (rL-RVG), and explore the effect of rL-RVG on proliferation and apoptosis of lung cancer cells.
A549 cells were infected with the rL-RVG and then detected for the expressions of RVG and NDV proteins by Western blotting. The cell proliferation was examined by MTT assay and apoptosis index and cell early apoptosis were respectively detected by TUNEL and annexin V-FITC/PI staining combined with flow cytometry. The expression of pro-apoptotic protein caspase-3 was observed by Western blotting. The LaSota strain of NDV was used as control group and PBS was the blank control.
Both RVG and NDV proteins were stably expressed in A549 cells infected with rL-RVG. MTT assay results showed that cell proliferation was significantly inhibited, and the inhibition rate of the rL-RVG group was higher than that of LaSota group. The apoptosis of A549 cells were promoted by rL-RVG infection. Flow cytometry revealed that the early apoptotic cells of the rL-RVG group increased as compared with the other two groups (P<0.05). Consistently, TUNEL assay showed the apoptotic index increased in the rL-RVG group as compared with the other two groups (P<0.05). Western blotting demonstrated that the expression of the pro-apoptotic protein caspase-3 was up-regulated. However, when we added specific broad-spectrum caspase inhibitor Z-VAD-FMK, the expression of the caspase-3 protein was obviously reduced.
The rL-RVG is stably expressed in the infected A549 cells. The rL-RVG inhibits lung cancer cell growth and promote cell apoptosis, and the effect of rL-RVG is better than the wild LaSota strain.
用表达狂犬病病毒糖蛋白（rL-RVG）的重组新城疫病毒（NDV）LaSota株疫苗感染肺腺癌A549细胞，探讨rL-RVG对肺癌细胞增殖和凋亡的影响。
用rL-RVG感染A549细胞，然后通过蛋白质免疫印迹法检测RVG和NDV蛋白的表达。采用MTT法检测细胞增殖情况，通过TUNEL法及Annexin V-FITC/PI染色结合流式细胞术分别检测凋亡指数和细胞早期凋亡情况。通过蛋白质免疫印迹法观察促凋亡蛋白caspase-3的表达。以NDV的LaSota株作为对照组，PBS作为空白对照。
RVG和NDV蛋白在感染rL-RVG的A549细胞中均稳定表达。MTT法检测结果显示细胞增殖受到显著抑制，且rL-RVG组的抑制率高于LaSota组。rL-RVG感染促进了A549细胞的凋亡。流式细胞术显示，与其他两组相比，rL-RVG组的早期凋亡细胞增多（P<0.05）。同样，TUNEL法检测显示，与其他两组相比，rL-RVG组的凋亡指数升高（P<0.05）。蛋白质免疫印迹法表明促凋亡蛋白caspase-3的表达上调。然而，当加入特异性广谱caspase抑制剂Z-VAD-FMK时，caspase-3蛋白的表达明显降低。
rL-RVG在感染的A549细胞中稳定表达。rL-RVG抑制肺癌细胞生长并促进细胞凋亡，且其效果优于野生型LaSota株。
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