血小板凝血酶蛋白1下调对肺腺癌A549细胞生物学功能的影响

Effects of thrombospondin 1 down - regulation on biological function of lung adenocarcinoma A549cells

目的观察血小板凝血酶蛋白1（THBS1）表达下调对肺腺癌A549细胞的细胞增殖、细胞凋亡和细胞周期的影响。 方法采用Western blot法检测肺腺癌细胞株A549、H1299、H358、H1975、PC9、PG49及Calu3中THBS1蛋白的表达水平，脂质体将THBS1小干扰RNA（siRNA）和对照siRNA分别转染至THBS1蛋白表达水平最高的A549细胞，Western blot检验转染THBS1 siRNA组、对照组和未转染组细胞中THBS1蛋白的表达，然后采用细胞计数试剂盒（CCK-8）检测各组细胞的增殖，采用流式细胞术分析各组细胞的细胞凋亡及细胞周期分布。 结果不同肺腺癌细胞株中THBS1蛋白的表达水平不同，其中A549细胞（1.120±0.087）表达最高（F＝358.087，P＝0.000）。与未转染组A549细胞和对照组比较：转染THBS1 siRNA组中A549细胞中THBS1蛋白表达量（0.053±0.023）显著降低（P＝0.000），细胞增殖能力减弱（P＝0.000），细胞的凋亡率[转染THBS1 siRNA组早期、中晚期及总凋亡率分别为（17.00±1.33）%、（3.70±0.56）%和（20.71±0.78）%升高（F值分别为115.899、14.418和167.329，P值分别为0.000、0.005和0.000]，G0/G1期细胞百分比（68.43±2.26）%显著升高（F＝46.521，P＝0.000），S期细胞百分比（22.33±2.15）%降低（F＝16.621，P＝0.004），G2/M期细胞百分比（9.23±2.35）%降低（F＝9.282，P＝0.015），以上差异均有统计学意义。 结论沉默THBS1基因，降低肺腺癌A549细胞中THBS1蛋白表达可以减弱细胞的增殖能力，促进细胞凋亡并抑制细胞周期进程。

ObjectiveTo observe the effect of down-regulation of platelet thrombospondin 1 （THBS1） on proliferation, apoptosis and cell cycle of lung adenocarcinoma A549 cells. MethodsTHBS1 protein expression in lung adenocarcinoma cell lines A549, H1299, H358, H1975, PC9, PG49 and Calu3 was detected by Western blotting. THBS1 small interfering RNA （siRNA） and control siRNA were transfected into A549 cells with the highest expression level of THBS1. The expression of THBS1 protein in THBS1 siRNA group, control group and transfected group was detected by Western blotting. Then cell counting kit-8 （CCK-8） kit was used to detect the proliferation of each group. Cell apoptosis and cell cycle distribution were analyzed by flow cytometry. ResultsThe expression levels of THBS1 protein in different lung adenocarcinoma cell lines were different, of which A549 cells had the highest expression （1.120±0.087; F=358.087, P=0.000）. Compared with non-transfected A549 cells and control group： The expression of THBS1 protein in A549 cells transfected with THBS1 siRNA （0.053±0.023） was significantly decreased （P=0.000）, the cell proliferation ability was decreased（P=0.000）, the apoptosis rates （THBS1 siRNA group： Early apoptosis rate, middle and late apoptosis rate and total apoptosis rate were （17.00±1.33）%, （3.70±0.56）% and （20.71±0.78）% were increased （F values were 115.899, 14.418 and 167.329 respectively, with P values of 0.000, 0.005 and 0.000 respectively）, the ratio of G0/G1 phase cells （68.43±2.26）% was significantly increased （F=46.521, P=0.000）, ratio of S phase cells （22.33±2.15）% was significantly increased（F=16.621, P=0.004） and G2/M phase cells （9.23±2.35）% decreased （F=9.282, P=0.015）, the difference was statistically significant. ConclusionSilencing THBS1 gene and decreasing the expression of THBS1 protein in lung adenocarcinoma A549 cells can weaken cell proliferation ability, promote cell apoptosis and inhibit the cell cycle progression.