上调GRIM-19基因对SPC-A1生长影响观察

Effects of stable expression of GR IM-19 on proliferation of lung adenocarcinoma cell line SPC-A1 growth

目的:探讨上调GRIM-19对肺腺癌细胞株SPA-A1细胞增殖的影响方法:电转染法将PIRES-GRIM19-MYC转染人肺腺癌SPC-A1细胞株,经G418筛选,得到稳定表达GRIM-19的SPC-A1细胞株,MTT法和裸鼠体内实验检测转染后SPC-A1细胞生长情况,RT-PCR和蛋白质印迹检测GRIM-19、STAT 3、Bcl-2、VEGF和MMP-2的表达。结果:成功建立了高表达GRIM-19的SPC-A1/GRIM-19细胞株,转染GRIM-19后SPC-A1细胞生长明显受抑制,其24、48、72h具体吸光度SPC/CON分别为1.07±0.01、1.32±0.15和1.81±0.18,SPC/GRIM-19分别为0.69±0.02、0.76±0.04和1.11±0.03,F=70.21,P=0.000 1。过表达GRIM-19显著抑制STAT 3、Bcl-2、VEGF和MMP-2的表达,其相对表达量分别为0.62±0.02、0.49±0.01、0.49±0.02和0.30±0.01,差异有统计学意义,t值分别为31.98、33.62、40.25和31.98,P值分别为0.000 9、0.000 7、0.000 4和0.000 9。SPC-A1/CON组BALB/C裸鼠的皮下肿瘤体积显著增大,SPC-A1/CON组BALB/C裸鼠的皮下肿瘤体积为(2.21±0.21)cm3,而SPC-A1/GRIM-19组BALB/C为(0.58±0.02)cm3,差异有统计学意义,t=30.23,P=0.000 9。结论:上调肺腺癌SPC-A1细胞中GRIM-19的表达对细胞的生长具有抑制作用。

OBJECTIVE： To explore the effects of expression of GRIM-19 on the proliferation of lung cancer cell lines SPC-A1. METHODS： The PIRES-GRIM19-MYC and PIRES-MYC were transfected into SPC-A1 cells by electropo- ration method,then the positive transfected cells were selected by G418. Proliferation inhibition of SPC-A1 were analyzed with dimethylthiazol-diphenyltetrazolium bromide （MTT）. SPC-A1/GRIM-19 and SPC-A1/CON were subcutaneously in- oculalted into nude mice and the growth of xenografted tumor was observed. The mRNA and protein expression of GRIM-19, STAT3, Bcl-2, VEGF, MMP-2 were identified by reverse transcription-polymerase chain reaction （RT-PCR） or Western blot. RESULTS： The transfencted SPC-A1/GRIM-19 cell line was estabilished successfully. The growth of SPC-A1/GRIM-19 cells,as indicated by MTT assay,was markedly inhibited on 24 h （1.07±0.01） vs （0.62±0.02） ,48 h （1.32±0.15） vs （0.76±0.04） and 72 h （1.81±0.18） vs （1.11±0.03） compared to the SPC-A1/CON cells （F= 70.21 ,P= 0. 000 1）. Upregulation of GRIM-19 can obviously down-regulate the expression of STAT3, BCL-2, VEGF, MMP-2 in lung adenocancinoma. Quantity of their protein relative expression were respectively 0.62±0.02,0.49±0.01, 0. 49±0. 02,0. 30±0. 01 compared to the SPC-A1/CON cells （t were 31.98,33.62,40.25 and 31.98;P were 0. 000 9, 0. 000 7,0. 000 4 and 0. 000 9, respectively）. Tumor volumes were significantly smaller in SPC-A1/GRIM-19 group （0.58±0.02） cm3 than that in SPC-A1/Control group （2.21±0.21） cm3 （t=30.23,P=0. 000 9）. CONCLUSION： Over- expression of GRIM-19 in SPC-A1 cells can inhibitthe proliferation ability in human lung adenocarcinoma cell.