网络首发:2015-01-20,
纸质出版:2015
移动端阅览
TGF-β1/Smad7信号通路在哈萨克族食管癌上皮间质转化中的作用[J]. 中山大学学报(医学科学版), 2015,36(1).
Activation of TGF-β1/Smad7 Signaling in Kazakh’s ESCC through Epithelial-Mesenchymal Transition[J]. Journal of Sun Yat-sen University (Medical Sciences), 2015, 36(1).
【目的】 探讨TGF-β1/Smad7信号通路在哈萨克族食管鳞癌上皮间质转化(EMT)中的作用及其机制。【方法】 运用免疫组织化学方法检测50例哈萨克族食管鳞癌组织及远端对应癌旁组织中TGF-β1与Smad7蛋白的表达。进一步在细胞水平,针对食管鳞癌细胞系Ec9706,基于RNA干扰技术,使用LipofectamineTM 2000试剂转染TGF-β1 siRNA。采用qRT-PCR技术检测TGF-β1及Smad7 mRNA表达水平;Western blot 技术检测转染后各组TGF-β1、Smad7以及EMT相关蛋白表达水平;MTT、划痕及流式细胞术等方法检测各组细胞转染后的增殖、迁移、凋亡及周期的变化。 【结果】 免疫组化检测结果显示,哈萨克族食管鳞癌组织中TGF-β1蛋白表达明显高于癌旁组织(P < 0.05);Smad7蛋白在食管鳞癌组织中的表达低于癌旁组织(P < 0.05)。转染TGF-β1 siRNA后,Ec9706细胞中TGF-β1 mRNA和蛋白表达量均降低,Smad7 mRNA和蛋白表达量增加;上皮标志物E-cadherin表达量增加,间质标志物Vimentin表达量降低,同时Ec9706细胞增殖、迁移受到抑制,细胞凋亡增加,细胞周期发生G0/G1期阻滞。【结论】 哈族食管鳞癌中存在TGF-β1/Smad7信号通路的激活,食管癌中TGF-β1高表达,抑制了Smad7的表达,从而促进了上皮间质转化过程,进一步促进了食管癌细胞的增殖和迁移,并抑制细胞凋亡,影响细胞周期中的G0/G1期。
【Objective】 To investigate the effects and mechanism of TGF-β1/Smad7 signaling pathway in Kazakh's esophageal squamous cell carcinoma (ESCC) through epithelial-mesenchymal transition (EMT). 【Methods】 Expression of TGF-β1 and Smad7 protein in carcinoma tissues and adjacent normal tissues of 50 Kazakh’s ESCC patients were detected by immunohistochemistry (IHC). Based on RNA interference technology, Ec9706 cell was transfected with TGF-β1 siRNA, using LipofectamineTM2000. The mRNA expression level of TGF-β1 and Smad7 were detected by qRT-PCR. Expression of TGF-β1, Smad7 and EMT-associated proteins were measured by Western blot after transfection. Then MTT, Wound-healing assay and flow cytometry were performed to detect cell’s proliferation, migration, apoptosis and cell cycle after transfection. 【Results】 The positive rate of TGF-β1 expression in carcinoma tissues of patients with ESCC was significantly higher than non-cancerous adjacent tissue (P < 0.05), while the positive expression rates of Smad7 in ESCC tissues was significantly lower than the non-cancerous adjacent tissues (P < 0.05). After transfected with TGF-β1 siRNA, expression of TGF-β1 and Vimentin were significantly decreased, meanwhile, expression of Smad7 and E-cadherin were increased. Furthermore, transfected with TGF-β1 siRNA inhibited cell proliferation, migration and promoted cell apoptosis.【Conclusion】 TGF-β1/Smad7 signaling activated in Kazakh's ESCC. TGF-β1 could promote EMT through inhibition of Smad7, consequently promote cell proliferation and migration, influence cell cycle at G0/G1 phase and inhibit the apoptosis of ESCC.
0
浏览量
392
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621
