MiR-1271靶向ZEB1抑制甲状腺乳头状癌细胞增殖和转移

MiR-1271 Targets ZEB1 to Inhibit the Proliferation and Metastasis of Papillary Thyroid Carcinoma Cells

[目的]探讨miR-1271对甲状腺乳头状癌细胞增殖和转移的影响及其作用机制。[方法]收集20例手术切除甲状腺乳头状癌患者的癌组织和对应的癌旁组织标本。采用RTqPCR检测组织和细胞系中miR-1271和ZEB1的表达水平;CCK-8检测K1细胞增殖活力;Transwell检测K1细胞侵袭和迁移能力;Western blot检测蛋白的表达水平;双荧光素酶报告基因验证miR-1271与ZEB1的靶向关系。[结果]miR-1271在甲状腺乳头状癌组织中的表达水平低于癌旁组织组织(1.175±0.534 vs 3.918±0.512,P<0.001)。与人甲状腺滤泡上皮正常细胞(0.985±0.062)相比,miR-1271在癌细胞系(TPC-1:0.752±0.052,K1:0.318±0.042,BCPAP:0.584±0.045)中的表达水平明显下调(P均<0.05),且在K1细胞中表达最低(P=0.016)。双荧光素酶报告基因结果显示,miR-1271通过结合ZEB1基因的3′UTR,进而抑制其表达水平。过表达ZEB1可明显缓解miR-1271对K1细胞增殖和转移的抑制作用。[结论]miR-1271通过靶向下调ZEB1的表达,进而抑制K1细胞增殖、侵袭、迁移和EMT。

[Objective]To explore the effect of miR-1271 on the proliferation and metastasis of papillary thyroid carcinoma cells and its underlying mechanism.[Methods]Twenty samples of cancer tissue and corresponding pericancerous tissues were collected from patients with papillary thyroid carcinoma at the Jiaxing Second Hospital from February 2016 to August 2018.RT-qPCR was used to detect the expression level of miR-1271 and ZEB1 in clinical samples and papillary thyroid carcinoma cell lines TPC-1,K1,BCPAP and normal human thyroid follicular epithelial Nthy-ori3-1 cells.The proliferation,invasion,and migration capacity of K1 cells were examined using CCK-8 assay and Transwell assay.Western blot was performed to measure protein level.Dual-luciferase reporter gene assay was applied to verify the interaction between miR-1271 and ZEB1 in papillary thyroid carcinoma.[Results]Expression of miR-1271 in the papillary thyroid carcinoma tissues was higher than that in the corresponding pericancerous tissues(1.175±0.534 vs 3.918±0.512,P<0.001).Compared with Nthy-ori3-1 cells(0.985±0.062),the expression of miR-1271 was down-regulated in papillary thyroid carcinoma cell lines(TPC-1:0.752±0.052,K1:0.318±0.042,BCPAP:0.584±0.045,all P<0.05).Overexpression of miR-1271 significantly suppressed the proliferation,invasion,migration and epithelial-mesenchymal transition(EMT).Dual-luciferase reporter gene assay results showed that miR-1271 decreased the expression of ZEB1 by binding to its 3′-untranslated region(UTR).Upregulation of ZEB1 attenuated the inhibitory effect of miR-1271 on the proliferation and metastasis of K1 cells.[Conclusion]miR-1271 can inhibit the proliferation,invasion,migration and EMT in papillary thyroid carcinoma cells by down-regulation of ZEB1 expression.