肿瘤相关成纤维细胞对大肠癌LoVo细胞奥沙利铂化疗敏感性的影响  

The effect of cancer-associated fibroblasts on the Oxaliplatin chemosensitivity of LoVo cells in colorectal cancer

作　　者：李明[1] 谭诗云[1] LI Ming;TAN Shiyun(Department of Gastroenterology, Renmin Hospital of Wuhan University, Hubei Key Laboratory of Digestive System Disease , Wuhan 430060, China)

机构地区：[1]武汉大学人民医院消化内科消化系疾病湖北省重点实验室,湖北武汉430060

出　　处：《胃肠病学和肝病学杂志》2019年第5期519-522,共4页Chinese Journal of Gastroenterology and Hepatology

基　　金：中央高校基本科研业务费专项基金(2042017kf0092)

摘　　要：目的观察肿瘤相关成纤维细胞(cancer-associated fibroblasts,CAFs)对大肠癌LoVo细胞奥沙利铂化疗敏感性的影响,并探讨其可能的作用机制。方法取大肠癌患者癌组织及癌旁正常结肠黏膜组织分别分离培养CAFs和正常成纤维细胞(normal fibroblasts,NFs),建立大肠癌LoVo细胞与CAFs或NFs共培养体系,奥沙利铂处理后CCK-8比色法检测LoVo细胞存活率,PI/Annexin V双染法检测细胞凋亡率;免疫印迹法(Western blotting)检测共培养体系中大肠癌细胞COX-2、上皮间质转化(EMT)相关蛋白(E-cadherin、Vimentin、Fibronectin)表达的改变,ELISA法检测共培养体系中细胞上清液中前列腺素E_2(PGE_2)含量。结果 CAFs可提高奥沙利铂处理后LoVo细胞存活率,并降低细胞凋亡率;LoVo细胞与CAFs共培养后,细胞COX-2蛋白表达上调,细胞上清液中PGE_2含量显著增加,且细胞EMT标志物蛋白E-cadherin表达降低,Vimentin、Fibronectin蛋白表达上调。结论 CAFs可降低大肠癌LoVo细胞对奥沙利铂的敏感性,其机制可能是通过上调大肠癌细胞COX-2表达和PGE_2合成,继而诱导EMT。Objective To observe the effect of cancer-associated fibroblasts(CAFs) on the Oxaliplatin chemosensitivity of LoVo cells in colorectal cancer, and to explore the possible mechanisms.Methods CAFs and normal fibroblasts(NFs) were isolated and cultured respectively from the tumor and adjacent normal colorectal tissue. After establishing co-cultured system and treated with Oxaliplatin, the cell survival was determined by the CCK-8 method. Apoptosis was detected by PI/Annexin V stainning. Western blotting was used for COX-2, E-cadherin, Vimentin and Fibronectin protein analysis. The expression of prostaglandin E2(PGE2) in the culture medium was detected by ELISA.Results Treatment with Oxaliplatin inhibited the cell viability and induced the cell apoptosis, and CAFs could inhibit the Oxaliplatin-induced cell viability and cell apoptosis of LoVo cells. Co-cultured with CAFs, the expressions of COX-2, Vimentin, Fibronectin, and PGE2 synthesis of LoVo cells were enhanced, and the expression of E-cadherin was decreased.Conclusion CAFs can reduce the Oxaliplatin chemosensitivity of LoVo cell, which is associated with a stimulation of epithelial-to-mesenchymal transition(EMT)via promotion of COX-2 expression and PGE2 synthesis.

关 键 词：肿瘤相关成纤维细胞 大肠癌 环氧合酶-2 奥沙利铂 化疗敏感性 

分 类 号：R735.3[医药卫生—肿瘤]