高尔基体基质蛋白130通过诱导上皮间充质转化促进甲状腺乳头状癌细胞TPC-1的侵袭和迁移

陈婕; 汤利华; 李正明

doi:10.13418/j.issn.1001-165x.2019.01.014

中国临床解剖学杂志 ›› 2019, Vol. 37 ›› Issue (1) : 64-70. DOI: 10.13418/j.issn.1001-165x.2019.01.014

实验研究

高尔基体基质蛋白130通过诱导上皮间充质转化促进甲状腺乳头状癌细胞TPC-1的侵袭和迁移

陈婕¹，　汤利华¹，　李正明²

作者信息 +

Golgi Matrix Protein 130 promotes papillary thyroid carcinoma cell TPC-1 invasion and migration through regulating epithelial-to-mesenchymal transition

CHEN Jie¹, TANG Li-hua¹, LI Zheng-ming²

Author information +

文章历史 +

摘要

目的　探究高尔基体基质蛋白130（Golgi Matrix Protein 130，GM130）对甲状腺乳头状癌细胞侵袭和迁移能力的作用及机制。方法　RT-PCR检测人甲状腺细胞Nthy-ori 3-1及甲状腺乳头状癌细胞BCPAP、K1、TPC-1中的GM130 mRNA水平。将细胞分为Control、scramble siRNA（si-scramble）、GM130 siRNA（si-GM130）3组。CCK8检测各组细胞增殖，划痕实验检测细胞迁移，Transwell检测细胞侵袭，Western Blot检测GM130、Ki67、增殖细胞核抗原（Proliferating cell nuclear antigen，PCNA）、基质金属蛋白酶-4（Matrix metalloproteinase-4，MMP-4）、基质金属蛋白酶-9（MMP-9）、钙依赖性粘附蛋白E（Calcium-dependent adhesion protein E，E-cadherin）、Snail、钙依赖性粘附蛋白N（N-cadherin）及波形蛋白Vinmentin的表达，同时利用免疫荧光检测Vimentin的表达。结果　GM130在TPC-1细胞中的mRNA水平最高，故利用TPC-1细胞进行后续实验。与Control组比较，si-GM130组中GM130表达显著下调，细胞增殖速率及Ki67、PCNA表达显著降低，细胞划痕闭合率显著下降，侵袭细胞数及MMP-4、MMP-9表达显著减少。此外，与Control组比较，si-GM130组中E-cadherin表达显著升高，Snail、N-cadherin及Vimentin表达显著降低。同时Vimentin荧光值在si-GM130组中显著降低。结论　GM130可通过诱导上皮间充质转化促进甲状腺乳头状癌细胞TPC-1的侵袭和迁移。

Abstract

Objective　To investigate the effect and mechanism of Golgi Matrix Protein 130(GM130) on the abilities of invasion and migration in papillary thyroid carcinoma cells.　Methods　The mRNA levels of GM130 in thyroid cell Nthy-ori 3-1 and papillary thyroid carcinoma cell BCPAP, K1 and TPC-1 were measured by RT-PCR. Cells were divided into Control, scramble siRNA(si-scramble), and GM130 siRNA(si-GM130) groups. CCK8 was performed to evaluate cell proliferation. Wound healing assay was performed to evaluate migration. Transwell was performed to evaluate invasion. The expression of GM130, Ki67, Proliferating cell nuclear antigen(PCNA), Matrix metalloproteinase-4(MMP-4), MMP-9, calcium-dependent adhesion protein E (E-cadherin), Snail, N-cadherin and Vimentin were measured by Western blot. Meanwhile, immunofluorescence assay was performed to evaluate the expression of Vimentin.　Results　GM130 in TPC-1 cells was higher expressed than in other papillary thyroid carcinoma cell lines. Therefore, TPC-1 cells were used for further study. Compared with the control group, the expression of GM130 was decreased in si-GM130 group, which resulted in the cell proliferation rate. And the expression of Ki67, and PCNA were decreased markedly. The wound closure rate was lower notably. The invasive cells and the expression of MMP-4, and MMP-9 declined significantly. Additionally, the expression of E-cadherin in si-GM130 group increased compared with that in control group, whereas the expression of Snail, N-cadherin and Vimentin decreased significantly. Meanwhile, the fluorescent value of Vimentin in si-GM130 group decreased significantly.　Conclusions　GM130 can promote papillary thyroid carcinoma cell TPC-1 invasion and migration through inducing epithelial-to-mesenchymal transition.

导出引用

陈婕,汤利华,李正明. 高尔基体基质蛋白130通过诱导上皮间充质转化促进甲状腺乳头状癌细胞TPC-1的侵袭和迁移[J]. 中国临床解剖学杂志. 2019, 37(1): 64-70 https://doi.org/10.13418/j.issn.1001-165x.2019.01.014

CHEN Jie, TANG Li-hua, LI Zheng-ming. Golgi Matrix Protein 130 promotes papillary thyroid carcinoma cell TPC-1 invasion and migration through regulating epithelial-to-mesenchymal transition[J]. Chinese Journal of Clinical Anatomy. 2019, 37(1): 64-70 https://doi.org/10.13418/j.issn.1001-165x.2019.01.014

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