Hemmatzadeh Maryam, Mohammadi Hamed, Babaie Farhad, Yousefi Mehdi, Ebrazeh Mehrdad, Mansoori Behzad, Shanehbandi Dariush, Baradaran Behzad
Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Immunology, School of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2018 Aug;8(3):437-445. doi: 10.15171/apb.2018.051. Epub 2018 Aug 29.
Snail-1 is a transcription factor, which takes part in EMT, a process related to the emergence of invasion and cancer progression. The purpose of this study was to evaluate the effect of Snail-1 silencing on the human esophageal squamous cell carcinoma cell line, namely TE-8, in vitro. In this study, transfection of Snail-1 specific siRNA was conducted into TE-8 cells. The relative mRNA expression levels of Snail-1, Vimentin, CXCR4 and MMP-9 and transcription levels of miR-34a and let-7a were investigated by quantitative Real-time PCR. Western blotting was carried out to evaluate the Snail-1 protein level. Migration assay of TE-8 cells was also performed following the presence or absence of Snail-1 specific siRNA. MTT and TUNEL assays were performed to evaluate cell viability after Snail-1 silencing. It was found that treatment of cancer cells with the Snail-specific siRNA effectively downregulated the expression of Snail-1 in both mRNA and protein levels, and vimentin, CXCR4, and MMP-9 in mRNA level. However, it elevated the transcript levels of miR-34a and let-7a expressions. Furthermore, transfection of cancer cells with the Snail-specific siRNA significantly induced apoptosis in TE8 cells. Moreover, suppression of Snail-1 led to diminished cell migration. It seems that Snail-specific siRNA can significantly interrupt esophageal cancer cell migration and reduce metastatic-related factors and induce miR-34a and let-7a in vitro. The bottom line is that therapeutic approaches via targeting Snail-1 can be used for ESCC treatment, suggesting that other possible target molecules for ESCC therapy require to be explored.
Snail-1是一种转录因子，它参与上皮-间质转化（EMT），这一过程与侵袭的出现和癌症进展相关。本研究的目的是在体外评估Snail-1沉默对人食管鳞状细胞癌细胞系TE-8的影响。在本研究中，将Snail-1特异性小干扰RNA（siRNA）转染到TE-8细胞中。通过定量实时聚合酶链反应（PCR）研究Snail-1、波形蛋白、趋化因子受体4（CXCR4）和基质金属蛋白酶-9（MMP-9）的相对信使核糖核酸（mRNA）表达水平以及微小核糖核酸-34a（miR-34a）和微小核糖核酸-7a（let-7a）的转录水平。进行蛋白质免疫印迹法以评估Snail-1蛋白水平。在存在或不存在Snail-1特异性siRNA的情况下，也对TE-8细胞进行迁移试验。进行噻唑蓝（MTT）和脱氧核糖核苷酸末端转移酶介导的缺口末端标记（TUNEL）试验以评估Snail-1沉默后的细胞活力。结果发现，用Snail特异性siRNA处理癌细胞可有效下调Snail-1在mRNA和蛋白质水平的表达，以及波形蛋白、CXCR4和MMP-9在mRNA水平的表达。然而，它提高了miR-34a和let-7a表达的转录水平。此外，用Snail特异性siRNA转染癌细胞可显著诱导TE8细胞凋亡。而且，Snail-1的抑制导致细胞迁移减少。似乎Snail特异性siRNA可在体外显著阻断食管癌细胞迁移并减少转移相关因子并诱导miR-34a和let-7a。关键在于，通过靶向Snail-1的治疗方法可用于食管鳞状细胞癌（ESCC）治疗，这表明需要探索ESCC治疗的其他可能靶分子。
