Lumican基因对人肺腺癌细胞株A549体外增殖和侵袭的影响

目的 观察基膜聚糖(Lumican)基因过度表达对人肺腺癌细胞株A549体外增殖和侵袭的影响.方法 以携带人Lumican基因的重组慢病毒感染A549细胞,用嘌呤霉素(puromycin)筛选法建立稳定细胞株.分别用Real-time PCR和Western blotting方法检测A549细胞组、空载体组及Lumican感染组目的基因mRNA和蛋白的表达.运用MTT法研究Lumican基因对A549细胞增殖的影响.运用Transwell法检测Lumican基因对A549细胞侵袭的影响.结果 ①各组Lumican mRNA表达差异显著(x2=21.60,P＜0.01),Lumican感染组明显高于其它两组(F=102.86,P＜0.000 1),Lumican感染组的蛋白表达水平高于其它两组.②Lumican感染组在不同时间(1、2、3、4、5 d)OD(吸光度)值与A549细胞组和空载体组差异不显著(P＞0.05).③各组穿膜细胞数差异显著(x2=23.49,P＜0.01),Lumican感染组明显高于其它两组(F=73.92,P＜0.001).结论 成功构建了Lumican高表达的人肺腺癌A549稳定细胞株,Lumican基因对人肺腺癌细胞的体外增殖能力无影响,但能增强人肺腺癌细胞的体外侵袭能力.

Effect of Lumican on Proliferation and Invasion of Human Lung Carcinoma Cell Line A549 in vitro

Objective To evaluate the effect of overexpression of Lumican on the proliferation and invasion of lung carcinoma cell line A549 in vitro. Methods The stable Sell lines were obtained by puromycin screening to select the A549 cells infected by recombinant lentiviral vector carrying human Lurnican gene. The mRNA expression level of Lumican gene in the A549 cell group, the empty vector group and the Lumican infection group were detected with Real-Time PCR respectively. The protein expression level of Lumican gene in the three groups were detec- ted with Western blotting method. The MTI＂ assay was used to investigate Lumican gene＇s effect on proliferation of the A549 ceils. The tran- swell method was used to detect it＇s effect on invasion of the A549 cells in vitro. Results （1）The expression level of mRNA in the each group was significantly different（ x2 =21.60,P 〈0. 01 ）. The expression level of mRNA in Lumican infection group was significantly higher than that in the other two groups（ F = 102. 86 ,P 〈0. 001 ）. The expression level of Lumican protein in Lumican infection group was higher than that in the other two groups. （2）There were no significant difference in OD（ Optical Density） values detected by MTT assay among the three groups at each time point after induction （P 〉 0. 05 ）. （3）The number of transfected ceils that passed the well in the each group was signifi- cantly different （ x2 = 23.49, P 〈 0. 01 ）. The number of transfected cells that passed the well in Lumican infection group was significantly higher than that in the other two groups（ F =73.92 ,P 〈0. 001 ）. Conclusions The A549 cell lines that stabely expressing Lumican is suc- cessfully established in the study. Lumican gene is not involved in proliferation of A549 cells, but it promotes their invasion in vitro.