摘要
目的探讨维生素D受体(VDR)对肠道肿瘤生长的影响以及与β-catenin信号通路之间的关系。方法体外培养的人类结肠癌SW480细胞株经维生素D处理4h,免疫沉淀法检测VDR和t3-catenin蛋白的交互作用,24h后用Westernblot检测细胞E.cadherin蛋白的表达。体内实验比较APCmin/+VDR-/-与APCmin/+小鼠,免疫组化法检测两型小鼠肠道肿瘤VDR、β-catenin和Brdu蛋白的表达,Westernblot检测肿瘤和肿瘤旁组织β-catenin蛋白的表达。结果维生素D处理SW480细胞后,VDR和β-catenin蛋白相互结合,随后E-cadherin蛋白表达增高(对照组灰度值145.57±4.21,实验组109.35±3.56,t=32.63,P〈0.05)。缺失VDR的APCmin/+VDR-/-小鼠肠道肿瘤中β-catenin蛋白表达免疫组化(灰度值140.51±2.57)及Westemblot(灰度值166.47±2.36)检测均高于APCmin/+肿瘤(分别为145.41±3.62、182.35±3.24,t=2.65,4.36,P〈0.05)。结论维生素D抑制肠道肿瘤增殖的作用可能通过VDR影响β-catenin信号通路来完成。
Objective To explore the effect of vitamin D receptor (VDR) in intestinal tumor development and the relationship between VDR and β-catenin signaling pathway. Methods The interaction of vitamin D receptor and β-catenin were detected by co-immunoprecipitation assay after human colonic car- cinoma cells SW480 were treated with vitamin D in vitro for 4 hours. The expression of E-cadherin protein was detected by Western blot after treated for 24 hours. To compare APCmin/+ VDR - / - and APCmin/ + mice in vivo, the expression of VDR, β-catenin and BrdU proteins in intestinal tumor were determined by immunohistochemistry. The expression of β-catenin protein in tumor and adjacency intestinal was further determined by Western blot. Results After SW480 cells were treated with vitamin D, vitamin D receptor and β-catenin protein showed binding, the expression of E-cadherin protein further increased ( Gray value the control group 145.57 + 4. 21, Gray value of the experimental group 109. 35± 3.56, t = 32. 63, P 〈 0. 05 ). Immunostaining and Western blot detection ( Gray value 166.47±2. 36) showed a marked increase of β- catenin level( Gray value 140. 51 ±2. 57 ) in APCmin/+ VDR - / - tumor compared to APCmin/+ tumor( 145.41±3.62,182. 35 ± 3.24,t = 2. 65,4. 36, P 〈 0. 05 ). Conclusions The role of vitamin D suppressing intestinal tumor may be achieved through VDR affectingβ-catenin signaling pathway.
出处
《中国医师杂志》
CAS
2011年第5期577-580,共4页
Journal of Chinese Physician
基金
国家自然科学基金项目(20972046)