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
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摘要
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 polysaccharide 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 groups and one group was the control group).The concentrations of safflower polysaccharide in the experimental groups were 0.16g/L,0.32g/L,and 0.64g/L respectively.The cells were cultured for 48 hours.Cells morphological were observed and MTT method was used to detect the OD of each induced group and control group on 12 h,24h,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 safflower polysaccharide on hela were calculated.Then cells apoptosis was detected by Annexin V-FITC/PI double labeling method.The expression of mRNA of VEGF,Nothc1,and Notch2 were detected by RT-PCR in each group on 48 hand the expression of MMP-9was detected by westening blotting.Finally,a comparative study was performed on each group.Results:The cell growth curve and proliferation inhibition rates had significant difference between the four groups(P<0.05).Itwas 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 apoptosis of human cervical cancer cells.The expression of VEGF,Notch1 and Notch2 in the three experimental groups were lower than those in the control group(P<0.05),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 apoptosis of cervical cancer cells and inhibit the proliferation of cervical cancer cells,which may be related to the VEGF and Notch signaling pathways.The mechanism of preventing the metastasis of cancer cells is related to the decrease the expression of MMP-9protein.
Objective:To study the effects of safflower polysaccharide 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 groups and one group was the control group).The concentrations of safflower polysaccharide in the experimental groups were 0.16g/L,0.32g/L,and 0.64g/L respectively.The cells were cultured for 48 hours.Cells morphological were observed and MTT method was used to detect the OD of each induced group and control group on 12 h,24h,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 safflower polysaccharide on hela were calculated.Then cells apoptosis was detected by Annexin V-FITC/PI double labeling method.The expression of mRNA of VEGF,Nothc1,and Notch2 were detected by RT-PCR in each group on 48 hand the expression of MMP-9was detected by westening blotting.Finally,a comparative study was performed on each group.Results:The cell growth curve and proliferation inhibition rates had significant difference between the four groups(P<0.05).Itwas 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 apoptosis of human cervical cancer cells.The expression of VEGF,Notch1 and Notch2 in the three experimental groups were lower than those in the control group(P<0.05),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 apoptosis of cervical cancer cells and inhibit the proliferation of cervical cancer cells,which may be related to the VEGF and Notch signaling pathways.The mechanism of preventing the metastasis of cancer cells is related to the decrease the expression of MMP-9protein.
引文
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[7]孙伟,童仕伦,郑勇斌,等.红花多糖对人结肠癌LoVo细胞增殖、凋亡、侵袭作用机制研究[J].安徽医药,2016,20(6):1045-1050.
[8]石亚利,江静.Notch信号通路在宫颈癌中的研究进展[J].中国妇产科临床杂志,2016,17(3):286-288.
[9]缪璐,陆晓媛.γ-分泌酶抑制剂DAPT抑制人宫颈癌HeLa细胞的增殖[J].徐州医学院学报,2010,31(6):406-408.
[10]夏美慧,谷爽,孙际童,等.姜黄素通过Notch途径抑制人宫颈癌细胞株HeLa细胞增殖[J].中国妇幼保健,2008,23(18):2588-2590.
[11]侯宇.宫颈癌组织中COX2、P21、MMP-9的蛋白表达及临床意义[J].中南医学科学杂志,2016,44(6):664-666.
[2]Tan KY,Liu CB,Chen AH,et al.The role of traditional Chinese medicine in colorectal cancer treatment[J].Tech Coloproctol,2008,12(1):1-6.
[3]杨婧,戚基萍,王锐,等.红花多糖对人宫颈癌Hela细胞增殖和VEGF表达的影响[J].中国实验方剂学杂志,2016,22(8):111-114.
[4]张晓莉,李玉婷,王亚贤,等.红花多糖的提取与含量测定[J].中国实验方剂学杂志,2010,16(7):19-21.
[5]王婷婷,石学魁,孙阳,等.红花多糖对人胃癌细胞SGC-7901凋亡的形态学实验研究[J].中医药信息,2015,32(2):19-21.
[6]孙阳,杨婧,张琪琪,等.红花多糖对肝癌细胞增殖阻滞的机制探讨[J].中国实验方剂学杂志,2014,20(13):156-159.
[7]孙伟,童仕伦,郑勇斌,等.红花多糖对人结肠癌LoVo细胞增殖、凋亡、侵袭作用机制研究[J].安徽医药,2016,20(6):1045-1050.
[8]石亚利,江静.Notch信号通路在宫颈癌中的研究进展[J].中国妇产科临床杂志,2016,17(3):286-288.
[9]缪璐,陆晓媛.γ-分泌酶抑制剂DAPT抑制人宫颈癌HeLa细胞的增殖[J].徐州医学院学报,2010,31(6):406-408.
[10]夏美慧,谷爽,孙际童,等.姜黄素通过Notch途径抑制人宫颈癌细胞株HeLa细胞增殖[J].中国妇幼保健,2008,23(18):2588-2590.
[11]侯宇.宫颈癌组织中COX2、P21、MMP-9的蛋白表达及临床意义[J].中南医学科学杂志,2016,44(6):664-666.