SCD1对甲状腺乳头状癌细胞增殖和侵袭的影响及其机制研究

Mechanism of SCD1 in promoting proliferation, migration and invasion of human papillary thyroid cancer cells

目的:探讨硬脂酰辅酶A去饱和酶1(stearoyl-CoA desaturase 1,SCD1)对甲状腺乳头状癌(papillary thyroid cancer,PTC)细胞TPC1增殖、迁移及侵袭的影响。方法:采用Western blot方法检测人甲状腺滤泡上皮正常细胞Nthy-ori 3-1和甲状腺乳头状癌细胞株TPC1、BCPAP和K1中SCD1的表达量。将阴性对照和SCD1 siRNA转染人甲状腺乳头状癌细胞TPC1,采用实时荧光定量PCR(qRT-PCR)及Western blot实验检测各组细胞中SCD1 mRNA及蛋白的表达水平;并通过CCK-8法和Transwell小室实验分别检测干扰SCD1对TPC1细胞增殖、迁移及侵袭的影响,同时通过qRT-PCR和Western blot实验检测各组细胞中上皮-间质转化(epithelial mesenchymal transition,EMT)相关蛋白:神经钙黏蛋白(N-cadherin)、Snail、波形蛋白(vimentin)和上皮钙黏蛋白(E-cadherin)的表达水平。结果:SCD1在TPC1中的表达水平明显高于Nthy-ori 3-1、BCPAP和K1(P<0.05);与空白对照组(Control)和对照干扰组(si-NC)相比,干扰SCD1组(si-SCD1)SCD1的mRNA和蛋白表达水平显著降低(P<0.05);干扰SCD1后TPC1细胞的增殖、迁移和侵袭明显被抑制(P<0.05);同时,干扰SCD1增加E-cadherin蛋白的表达,但抑制vimentin、Snail和N-cadherin蛋白的表达(P<0.05)。结论:干扰SCD1可通过抑制细胞的EMT抑制甲状腺乳头状癌细胞TPC1的增殖、迁移及侵袭能力。

Objective:To investigate the effects of SCD1 on the proliferation,migration and invasion of human papillary thyroid cancer cell line TPC1. Methods:The expression of SCD1 in human thyroid follicular epithelial normal cells Nthy-ori 3-1 and thyroid papillary cancer cell lines TPC1,BCPAP and K1 were detected by Western blot assay. The negative control and SCD1 si RNA were transfected respectively into human thyroid papillary carcer cell line TPC1,then the expressions of SCD1 m RNA and protein in each group were tested by real-time fluorescence quantitative PCR(q RT-PCR)and Western blot assay.The effects of interfering SCD1 on the proliferation,migration and invasion of TPC1 cells were detected by CCK-8 method and Transwell chamber assay,respectively. Meanwhile,the expression levels of epithelial-mesenchymal transition(EMT)related proteins(N-cadherin,Snail,vimentin and E-cadherin)were detected by q RT-PCR and Western blot. Results:The expression level of SCD1 in TPC1 was significantly higher than those in Nthy-ori 3-1,BCPAP and K1(P<0.05). Compared with the blank control group(Control)and the control interference group(si-NC),the expression of SCD1 m RNA and protein in the interference SCD1 group(si-SCD1)were significantly decreased(P<0.05). The proliferation,migration and invasion of TPC1 cells were significantly inhibited after interfering with SCD1(P<0.05). It was also found that interfering SCD1 could increase the expression of E-cadherin protein while inhibited the expression of vimentin,Snail and N-cadherin protein significantly(P<0.05).Conclusion:Interfering SCD1 could inhibit the proliferation,migration and invasion of thyroid papillary cancer cell TPC1 by inhibiting the EMT procedure.