Zhou Kun, Ge Mingjian
Department of Cardiothoracic Surgery, First Affiliated Hospital, Chongqing Medical University, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 May;29(5):492-5, 497.
To investigate the effect of the lumican gene on the proliferation of lung adenocarcinoma cell A549 and the possible mechanism.
The lung adenocarcinoma cell line A549 was infected with the recombinant lentivirus carrying the human lumican gene and a stable cell line was obtained via puromycin selection. The expression of lumican mRNA and protein was confirmed by fluorescence quantitative PCR(FQ-PCR) and Western blotting. We examined the cell growth curve, doubling time, proliferation index using MTT assay and flow cytometry and observed the status of the tumor in the experimental animals. Meanwhile, Western blotting was used to detect the expressions of RhoC and p-Akt proteins.
The A549 cell line with the lumican gene over-expression was successfully built. Compared with the control group and the empty vector group, the cells in the experimental group grew faster (P<0.05), the doubling time was shortened (P<0.05), the proliferation index went up (P<0.05), subcutaneous tumors in volume and mass increased and cells of the mitotic figures became more (P<0.05), and the expressions of RhoC and p-Akt proteins were raised (P<0.05). However, there was no obvious difference between the control group and the empty vecor group (P>0.05).
Lumican gene can promote the proliferation of lung adenocarcinoma cell A549, and its mechanism may be related to increased RhoC and p-Akt protein expressions.
探讨富含亮氨酸蛋白聚糖基因对肺腺癌细胞A549增殖的影响及其可能机制。
用携带人富含亮氨酸蛋白聚糖基因的重组慢病毒感染肺腺癌细胞系A549,经嘌呤霉素筛选获得稳定细胞系。采用荧光定量PCR(FQ-PCR)和蛋白质印迹法检测富含亮氨酸蛋白聚糖mRNA和蛋白的表达。用MTT法和流式细胞术检测细胞生长曲线、倍增时间、增殖指数,并观察实验动物肿瘤状态。同时,用蛋白质印迹法检测RhoC和p-Akt蛋白的表达。
成功构建了富含亮氨酸蛋白聚糖基因过表达的A549细胞系。与对照组和空载体组相比,实验组细胞生长加快(P<0.05),倍增时间缩短(P<0.05),增殖指数升高(P<0.05),皮下肿瘤体积和质量增加,有丝分裂细胞增多(P<0.05),RhoC和p-Akt蛋白表达升高(P<0.05)。而对照组和空载体组之间无明显差异(P>0.05)。
富含亮氨酸蛋白聚糖基因可促进肺腺癌细胞A549的增殖,其机制可能与RhoC和p-Akt蛋白表达增加有关。