半枝莲多糖联合R568抑制宫颈癌Hela细胞增殖作用的研究

A study on SBP plus R568 inhibit the proliferation of cervical cancer Hela cells

目的:观察半枝莲多糖(Sculellaria Barbata Polysaccharides,SBP)联合R568[钙敏感受体(Calcium-sensing receptor,CaSR)激动剂]抑制宫颈癌Hela细胞增殖情况并分析其机制。方法:60只裸鼠随机分为模型组(移植瘤模型),模型+R568组,模型+Calhex231(CaSR抑制剂)组,模型+SBP组,模型+SBP+R568组和模型+SBP+Calhex231组。Hela细胞随机分为对照组,R568组,Calhex231组,SBP组,SBP+R568组和SBP+Calhex231组。分析比较各组移植瘤重量、体积和抑瘤率变化;酶联免疫吸附测定(Enzyme Linked Immunosorbent Assay,ELISA)检测裸鼠血清和Hela细胞培养基中基质金属蛋白酶(Matrix Metalloproteinase,MMP)2和MMP9表达水平;细胞计数试剂盒-8(Cell Counting Kit-8,CCK8)检测Hela细胞增殖情况;流式细胞术检测Hela细胞凋亡情况;Western blot检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、兔抗人单克隆抗体(Bax)、E-钙黏蛋白(E-cadherin)和β-连环蛋白(β-catenin)的蛋白表达量变化;荧光定量-聚合酶链式反应(qRT-PCR)检测Bcl-2和Bax基因表达情况。结果:移植瘤模型中,与模型组相比较,模型+R568组,模型+SBP组和模型+SBP+R568组裸鼠血清中MMP2和MMP9含量显著降低,抑瘤率显著增加;Bax蛋白表达增加,而Bcl-2表达降低。细胞实验中,与对照组相比较,R568组,SBP组和SBP+R568组Hela细胞增殖速度显著降低,细胞凋亡水平显著增加,培养基中MMP2和MMP9含量水平显著下降;E-cadherin和β-catenin蛋白增加;Bax基因表达增加,Bcl-2基因表达降低。在Calhex231组和SBP+Calhex231组上述结果相反。结论:半枝莲多糖联合R568可抑制Hela细胞增殖并促进其凋亡。

Objective:To observe the inhibition of Sculellaria barbata polysaccharides plus R568(CaSR agonist)on the proliferation of cervical cancer Hela cells and analyze its mechanism.Methods:60 nude mice were randomly divided into the model group(xenograft tumor model),model+R568 group,model+Calhex231(CaSR inhibitor)group,model+SBP group,model+SBP+R568 group and model+SBP+Calhex231 group.Hela cells were randomly divided into the control group,R568 group,Calhex231 group,SBP group,SBP+R568 group and SBP+Calhex231 group.The changes in the weight,volume and tumor inhibition rate of transplanted tumors in each group were analyzed and compared.Results:In the transplanted tumor model,compared with the model group,the serum levels of MMP2 and MMP9 in nude mice of the model+R568 group,the model+SBP group and the model+SBP+R568 group were significantly reduced,and the tumor inhibition rate was significantly increased.The expression of Bax protein was increased,while the expression of Bcl-2 was decreased.In the cell experiment,compared with the control group,the proliferation rate of Hela cells in the R568 group,SBP group and SBP+R568 group was significantly reduced,the level of apoptosis was significantly increased,and the levels of MMP2 and MMP9 in the culture medium were significantly decreased;E-cadherin andβ-Catenin protein were increased.Bax gene expression was increased,Bcl-2 gene expression was decreased.In the Calhex231 group and the SBP+Calhex231 group,the above results are opposite.Conclusion:Scutellaria barbata polysaccharide with R568 can inhibit the proliferation of Hela cells and promote their apoptosis.