CaSR在宫颈癌C-33A细胞增殖和迁移中的作用研究  

The Role of Calcium-sensitive Receptor in Cervical Carcinoma C-33A Cell Proliferation and Migration

作　　者：袁辉 张淑敏[1] 王得利 栾海蓉 石屹[1] 徐吉雨 YUAN Hui;ZHANG Shumin;WANG Deli;LUAN Hairong;SHI Yi;XU Jiyu(Mudanjiang Medical University,School of Stomatology,Mudanjiang 157011,China;不详)

机构地区：[1]牡丹江医学院口腔医学院,黑龙江牡丹江157011 [2]牡丹江医学院基础医学院 [3]牡丹江医学院医学影像学院

出　　处：《中国医学创新》2020年第35期21-26,共6页Medical Innovation of China

基　　金：2018年度黑龙江省省属高等学校基本科研业务费科研项目(2018-KYYWFMY-0043);牡丹江市科学技术计划项目(Z2018s040)。

摘　　要：目的:观察钙敏感受体(calcium sensing receptor,CaSR)在宫颈癌C-33A细胞中的表达情况并分析其参与细胞增殖和迁移的机制。方法:30只裸鼠随机分为Model(移植瘤模型)组、Model+R568(CaSR激动剂)组、Model+Calhex231(CaSR抑制剂)组,每组10只。C-33A细胞分为Control组、R568组和Calhex231组。分析各组移植瘤体积和抑瘤率变化;ELISA检测裸鼠血清和C-33A细胞培养基中MMP2和MMP9含量变化;Western blot测定瘤组织和细胞中CaSR、Bax、Bcl-2、E-cadherin、β-catenin和VEGFR3表达变化;CCK8检测C-33A细胞增殖情况;细胞划痕实验检测C-33A细胞迁移;qRT-PCR检测Bcl-2和Bax基因表达情况。结果:30 d后取各组裸鼠瘤组织称重,Model+R568组的移植瘤瘤重低于Model组(P<0.05)。饲养5、10、15、20、25、30 d,测量移植瘤体积,与Model组比较,Model+R568组显著抑制肿瘤生长而Model+Calhex231肿瘤体积显著增加(P<0.05)。与Model组比较,Model+R568组裸鼠血清中MMP2和MMP9含量均显著降低(P<0.05)。Western blot结果显示,与Model组比较,Model+R568组的CaSR和E-cadherin蛋白表达均显著增加而Bcl-2的表达量显著减少(P<0.05)。细胞实验中,与Control组比较,R568组C-33A细胞增殖和迁移速度均显著降低(P<0.05),CaSR、Bax、E-cadherin和β-catenin表达均显著上调,而Bcl-2和VEGFR3表达均下调(P<0.05),不同时间点细胞内Ca^2+荧光强度显著增强(P<0.05),培养基中MMP2和MMP9含量水平均显著下降(P<0.05),Bax基因表达明显上调而Bcl-2表达下调(P<0.05)。在Calhex231组上述结果均相反。结论:CaSR激活能够抑制C-33A细胞增殖和迁移,并促进其凋亡。Objective:To observe the role of CaSR expression in cervical cancer C-33A cells and analyze its mechanism of involving in proliferation and migration.Method:30 nude mice were randomly divided into Model group,Model+R568(CaSR agonist)group and Model+Calhex231(CaSR inhibitor)group,10 nude mice in each group.C-33A cells were divided into control group,R568 group and Calhex231 group.The tumor volumes and inhibition rates of each group were analyzed;the MMP2 and MMP9 concentrations in nude mice serum and cell medium were detected by ELISA kits,and the expressions of CaSR,Bax,Bcl-2,E-cadherin,β-catenin and VEGFR3 in tumor tissue and cells were detected by Western blot;the proliferation and migration rates of C-33A cells were determined by CCK8 kits and cell scratch test,respectively.Bcl-2 and Bax gene expressions were detected by qRT-PCR.Result:After 30 days,the tumor tissues of each group of nude mice were weighed,and the tumor weight of the transplanted tumor in the Model+R568 group was lower than that of the Model group(P<0.05).After 5,10,15,20,25 and 30 days of feeding,tumor volume was measured,compared with the Model group,tumor growth was significantly inhibited in the Model+R568 group and tumor volume increased in the Model+Calhex231 group(P<0.05).Compared with the Model group,the levels of MMP2 and MMP9 in the Model+R568 group were significantly decreased(P<0.05).Western blot results showed that CaSR and E-cadherin protein expression were significantly increased in the Model+R568 group and Bcl-2 expression was significantly decreased in the Model+R568 group compared with the Model group(P<0.05).In cell experiments,the proliferation and migration rates of C-33A cells in group R568 were significantly reduced compared with those in the Control group(P<0.05);the expressions of CaSR,Bax,E-cadherin andβ-catenin were significantly up-regulated,while the expressions of Bcl-2 and VEGFR3 were down-regulated(P<0.05);the Ca^2+fluorescence intensity was significantly increased at different time points(P<0.05);the content

关 键 词：CASR C-33A细胞 MMP2 MMP9 凋亡 迁移 

分 类 号：R737.33[医药卫生—肿瘤]