枸杞多糖对人肺腺癌A549细胞增殖和凋亡的影响

Function of Lycium Barbarum Polysaccharide on Proliferation and Apoptosis of Human Lung Cancer A549 Cells

目的探讨枸杞多糖(LBP)对体外培养的人肺腺癌细胞A549的增殖抑制作用及其可能的作用机制。方法用不同浓度的LBP处理A549细胞,MTT法检测24、48、72 h时间点LBP对A549细胞的生长抑制率,实验设为对照组和实验组(1/2IC_(50)作用48小时),MTT法绘制生长曲线、细胞计数计算倍增时间、流式细胞仪检测凋亡率及其细胞周期、RT-PCR检测Survivin mRNA的变化、Western blot检测CyclinB1蛋白的变化,transwell体外侵袭实验观察药物对细胞体外侵袭的影响。结果 MTT显示不同浓度的LBP均能明显抑制A549细胞的增殖且成剂量-效应关系,实验组细胞的倍增时间、凋亡率与对照组相比,均有统计学意义(P<0.05);LBP使细胞阻滞在G_2期,Survivin mRNA表达和CyclinB1蛋白的表达均降低,与对照组相比差异有显著性(P<0.05)。结论 LBP可抑制A549细胞的增殖,其机理可能与LBP使Survivin mRNA表达下降引起细胞凋亡及CyclinB1蛋白的表达降低造成细胞周期阻滞及抑制细胞的侵袭能力有关。

Objective To investigate the growth inhibition and proliferation of human lung adenocarcinoma A549 cells induced by Lycium barbarum polysaccharide（LBP） in vitro,and the possible underlying mechanisms. Methods A549 cells cultured in vitro were divided into control group and experimental group （ 1/2IC50 for 48 hours） by the different concentrations of LBP treatment. The inhibitory effects of LBP on proliferation of A549 were determined by MYr assay at 24 h ,48 h ,72 h after the addition of LBP to A549 culture. Growth curve were generated by MTT assay, doubling time were calculated by cell counting, Cell cycle and apoptosis were examined by Flow cytometry （FCM）, Survivin mRNA changes were detected by RT-PCR, CyclinB1 protein changes were detected by Western blot, Transwell assay in vitro was utilized to evaluate the invasive activity. Results MTr assay demonstrated that different concentrations of LBP significantly inhibited the proliferation of A549 cells in a dase-dependent manner,the doubling time and the rate of apoptosis of cells in experimental group is dramatically different from the control group. （P 〈 0. 05 ）, It was found that LBP arrested A549 cells at G2 phase;Survivin mRNA expression and CyclinB1 protein expression were lower, compared with that in the control group （P 〈 0.05 ）. Conclusion The present study suggests that LBP can significantly inhibit the proliferation of A549 cells. The mechanism may be related to an arrest effect on cell cycle,reducing Survivin mRNA and CyclinB1 protein expression and inhibiting tumour invasion.