红花多糖对人宫颈癌Hela细胞增殖、凋亡及侵袭作用机制研究

Mechanism Research of Effects of Safflower Polysaccharide on Proliferation,Apoptosis and Invasion of Human Cervical Cancer Hela Cells

目的:研究红花多糖对人宫颈癌细胞(hela)体外增殖、凋亡及侵袭蛋白表达的影响。方法:将宫颈癌细胞分为4组(3组为实验组,1组为对照组),3个实验组红花多糖质量浓度分别为0.16、0.32、0.64g/L,连续干预培养48h,进行形态学观察,并采用MTT法检测各诱导组与对照组第12、24、36、48h的OD值,绘制各组细胞生长曲线,并计算不同浓度红花多糖对hela的增殖抑制率。采用Annexin VFITC/PI双标法检测48h各组细胞凋亡率,采用RT-PCR检测各组中VEGF,Notch1,Notch2的表达,westening blotting检测各组MMP-9的表达,进行各组对比研究。结果:4组间的细胞增殖曲线、增殖抑制率有明显差异(P<0.05),明确红花多糖能抑制人宫颈癌细胞的生长、増殖,呈现时间和浓度效应关系。流式细胞仪分析结果提示红花多糖促进人宫颈癌细胞凋亡。VEGF、Notch1、Notch2mRNA在3组实验组的表达量较对照组低(P<0.05),MMP-9蛋白在3组实验组的表达较对照组低(P<0.05)。结论:红花多糖能促进宫颈癌细胞凋亡,可抑制宫颈癌细胞增殖,其作用机制可能与VEGF、Notch信号通路有关,抑制癌细胞转移机制与MMP-9蛋白表达减少有关。

Objective： To study the effects of safflower polysaccliaride on proliferation, apoptosis and invasion protein expression of cervical cancer cells （Hela） in vitro. Methods： Cervical cancer cells were divided into four groups （Three groups were experimental groupxs and one group was the control group）. The concentratioas of safflower polysaccharide in the experimental groups were 0. 16 g/L,0. 32 g/L？ and 0. 64 g/ L respectively. The cells were cultured for 48 hours. Cells morphological were obsen^ed and MTT method was used to detect the （）D of each induced group and control group on 12 h,24 h,36 h,and 48 h. According to the results of OD of each group,growth curve were drawn and the inhibition rates of the proliferation of different concentrations of s〈ifflower polys〈iccharide on hela were calculated. Then cells apoptosis was detected by Annexin V-FITC/PI double labeling method The expression of mRNA of VEGF＇, Nothcl ？ and Notch2 were detected by RT-PGR in each group on 48 h and the expression of MMP-9 was detected by westening blotting. Finally,a comparative study was performcxl on each group. Results：The cell growth curve and proliferation inhibition rates had significant difference between the four groupsCP^O. 05）. It was clear that safflower polysaccharide could inhibit the growth and proliferation of human cervical cancer cells？ whose effects were associated with concentration and time. The further results of flow cytometry showed that safflower polysaccharide could promote the a[X）ptosis of human cervical cancer cells. The expression of VEGF？Notchl and Notch2 in the three experimental groups werelower than those in the control grou p （P 〈 0 . 0 5 ） ,and the expression of MMP-9 protein in the experimental group was lower than that in the control group（P〈0 . 05）. Conclusion：Safflower polysaccharide can promote the ajx）ptosis of cervical cancer cells and inhibit the proliferation of cenncal cancer cells,which may be related to the VEGF and Notch signaling pathways. The meclianism of preventing the metastasis of cancer cells is related to the decrease the expression of MMP-9 protein.