人呼吸道合胞病毒M2-1基因转染对人肺腺癌细胞生长特性的影响

Influence of transfection with human respiratory syncytial virus M2-1 gene on growth characteristics of human lung adenocarcinoma cell lines PAa and A549

目的探讨人呼吸道合胞病毒(hRSV)M21基因转染对人肺腺癌细胞体外生长特性的影响。方法应用脂质体转染法,将携带M21基因的重组载体pXJ41/M21转入人肺腺癌PAa和A549细胞,并获稳定表达;以不含M21基因的空白载体为对照,采用流式细胞术、MTT法、胰酶消化试验、软琼脂集落形成实验和Boyden小室等方法观察转染细胞的生长特性、贴壁能力及侵袭能力变化。结果与转染前和转染空白质粒组比较,转染M21基因的PAa和A549细胞的G2/M期细胞比例均显著增加(P<0.05,P<0.01);细胞增殖明显加快(P<0.01),倍增时间缩短;集落形成率显著增加(P<0.01);PAa细胞胰酶消化离壁时间缩短(P<0.01),而A549细胞在3组间无统计学差异;Boyden小室实验中,A549细胞透膜数显著增加(P<0.01),而PAa细胞因透膜数很少,均无法计数。结论M21基因转染能提高体外培养的PAa和A549细胞的增殖能力和侵袭能力,感染hRSV病毒的肺癌患者应引起重视。

Objective:To study the influence of human respiratory syncytial virus (hRSV) M2-1 gene on human lung adenocarcinoma cell lines PAa and A549 in vitro. Methods: Liposome transfection method was used to transfect the recombined pXJ-41/M2-1 into human lung adenocarcinoma PAa and A549 cells, and PAa and A549 cells stably expressing M2-1 gene were acquired and verified. PAa and A549 cells were also transfected with blank plasmid. Changes of proliferation speed, adhesion ability to the culture utensil and invasive ability were studied by MTT, flow cytometry, trypsin digestion and Boyden Chamber. Results: Compared with those of non-transfection and blank plasmid-transfected PAa and A549 cells, the G_2/M period cell rate in M2-1 transfected group increased significantly (P<0.01,P<0.05); the proliferation ability increased (P<0.01) and the double-augment period shortened; the clone forming rate increased(P<0.01); the period for PAa trypsin digestion completely leaving from the culture bottle bottom decreased (P<0.01), while there was no statistical change in A549 cells; in Boyden Chamber experiment, the quantity of the A549 cells breaking through the matrigel cell increased (P<0.01), while the quantity of PAa was too little to be counted. Conclusion: The M2-1 gene of hRSV can promote the proliferation and invasive ability of the transfected PAa and A549 cell lines in vitro. Lung cancer patients with hRSV infection deserve our attention.