摘要
目的探讨TGF-β/Smad2信号通路在锰-双希夫碱复合物(Salen-Mananese,Salen-Mn)抑制肾细胞癌侵袭转移中的作用。方法 MTT法检测Salen-Mn对肾细胞癌786-O细胞的细胞活性影响。用伤口愈合实验和Transwell侵袭迁移实验检测Salen-Mn对肾细胞癌786-O细胞的侵袭迁移能力。用Western blot以及real-time PCR检测Salen-Mn对肾细胞癌786-O细胞内上皮间质转化相关蛋白标志物(E-cadherin、N-Cadherin、Vimentin和Snail)的表达。通过Western和siRNA转染技术检测TGF-β/Smad2信号通路在Salen-Mn对EMT调控中的作用。结果 Salen-Mn可以抑制肾细胞癌786-O细胞的增殖和侵袭迁移(P<0.05)。与此同时,Salen-Mn能在mRNA和蛋白水平上调E-cadherin的表达,并下调N-Cadherin、Vimentin和Snail的表达(P<0.05)。Western blot结果显示,Salen-Mn能够抑制肾细胞癌786-O细胞内TGF-β和p-Smad2的表达,且TGF-β的过表达能够逆转Salen-Mn对EMT和侵袭迁移的抑制。结论 Salen-Mn通过下调TGF-β/Smad2信号通路抑制肾细胞癌的侵袭转移,这为肾细胞癌的临床治疗提供了一定的理论依据。
Objective To elucidate the role of TGF-β/Smad2 signaling in anti-metastatic effect of salen-Mn on renal cell carcinoma cells. Methods MTT was used to detect the cell viability of renal cell carcinoma 786-O cells after salen-Mn treatment. Wound healing assay and Transwell assays were performed to detect the effect of salen-Mn on migration and invasion of renal cell carcinoma cells.Western blot and real-time PCR were used to detect the protein and mRNA levels of E-cadherin,N-Cadherin,Vimentin and Snail in renal cell carcinoma 786-O cells treated with salen-Mn. Western blot and plasmid transfection were performed to detect the function of TGF-β/Smad2 signaling in the salen-Mn-regulated EMT. Results Salen-Mn inhibited the proliferation,migration and invasion of human renal cell carcinoma 786-O cells（ P〈0. 05）. Salen-Mn upregulated E-cadherin expression,but downregulated N-Cadherin,Vimentin and Snail in mRNA and protein levels（ P〈0. 05）. The results of Western blot revealed that salen-Mn downregulated the expression of TGF-β and p-Smad2,and the overexpression of TGF-β reversed high expression of E-cadherin,low expression of Vimentin,and the inhibitory effect of migration and invasion induced by salen-Mn. Conclusion These findings suggest that salen-Mn could inhibit migration and invasion of renal cell carcinoma cells by negatively regulating TGF-β/Smad2 signaling pathway.
作者
寇博
寇青山
刘伟
KOU Bo;KOU Qingshan;LIU Wei(Department of Cardiovascular Surgery, First Affiliated Hospital of Medical School of Xi＇an Jiaotong University, Xi＇ an 710061, China;Physical Examination Center of First People＇ s Hospital of Xianyang;Department of Vascular Surgery, First Affiliated Hospital of Medical School of Xi ＇ an Jiaotong University)
出处
《山西医科大学学报》
CAS
2018年第4期352-356,共5页
Journal of Shanxi Medical University
基金
国家自然科学基金青年基金项目(81602562)
