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

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

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

1.重庆市第九人民医院内分泌科，重庆 400700； 2.华西医院内分泌科，成都 610041

收稿日期:2018-10-12 出版日期:2019-01-25 发布日期:2019-02-20
通讯作者: 汤利华，主治医师， E-mail：longding459938@sina.com
作者简介:陈婕(1984-)，硕士，主治医师，主要从事内分泌研究工作，Tel:13883319774，E-mail：xiubao12984925@sina.com
基金资助:
重庆市卫生和计划生育委员会医学科研项目(2016MSXM145)

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

1.Department of Endocrine, Chongqing Ninth People's Hospital, Chongqing 400700, China; 2.Department of Endocrinology, West China Hospital, Chengdu 610041, China

Received:2018-10-12 Online:2019-01-25 Published:2019-02-20

摘要/Abstract

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。

关键词: GM130, 　上皮间充质转化, 　甲状腺乳头状癌, 　侵袭, 　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.

Key words: GM130, 　Epithelial-to-mesenchymal Transition, 　Papillary thyroid carcinoma, Invasion, 　Migration

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.

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.

参考文献

[1] Xhaard C, Rubino C, Cléro E, et al. Menstrual and reproductive factors in the risk of differentiated thyroid carcinoma in young women in France: a population-based case-control study[J]. Am J Epidemiol, 2014, 180(10): 1007-1017.
[2] van Staalduinen J, Baker D, Ten Dijke P, et al. Epithelial mesenchymal transition inducing transcription factors: new targets for tackling chemoresistance in cancer[J]. Oncogene, 2018.
[3] Ko JH, Nam D, Um JY, et al. Bergamottin suppresses metastasis of lung cancer cells through abrogation of diverse oncogenic signaling cascades and epithelial-to-mesenchymal transition[J]. Molecules, 2018, 23(7): 1601.
[4] Shimizu D, Saito T, Ito S, et al. Overexpression of FGFR1 promotes peritoneal dissemination via epithelial-to-mesenchymal transition in gastric cancer[J]. Cancer Genomics Proteomics, 2018, 15(4): 313-320.
[5] Rahimian A, Barati G, Mehrandish R, et al. Inhibition of histone deacetylases reverses epithelial-mesenchymal transition in triple-negative breast cancer cells through a slug mediated mechanism[J]. Mol Biol, 2018, 52(3): 474-481.
[6] Duan L, Ye L, Zhuang L, et al. VEGFC/VEGFR3 axis mediates TGFβ1-induced epithelial-to-mesenchymal transition in non-small cell lung cancer cells[J]. Plos One, 2018, 13(7): e0200452.
[7] Li L, Sun P, Zhang C, et al. MiR-98 modulates macrophage polarization and suppresses the effects of tumor-associated macrophages on promoting invasion and epithelial-mesenchymal transition of hepatocellular carcinoma[J]. Cancer Cell Int, 2018, 18(1): 95.
[8] Fukagawa D, Sugai T, Osakabe M, et al. Protein expression patterns in cancer-associated fibroblasts and cells undergoing the epithelial-mesenchymal transition in ovarian cancers[J]. Oncotarget, 2018, 9(44): 27514-27524.
[9] Vasko V, Espinosa AV, Scouten W, et al. Gene expression and functional evidence of epithelial-to-mesenchymal transition in papillary thyroid carcinoma invasion[J]. Proc Natl Acad Sci U S A, 2007, 104(8): 2803-2808.
[10] Han F, Liu C, Zhang L, et al. Globozoospermia and lack of acrosome formation in GM130-deficient mice[J]. Cell Death Dis, 2017, 8(1): e2532.
[11] Zhao J, Yang C, Guo S, et al. GM130 regulates epithelial-to-mesenchymal transition and invasion of gastric cancer cells via snail[J]. Int J Clin Exp Pathol, 2015, 8(9): 10784-10791.
[12] Buschman MD, Rahajeng J, Field SJ. GOLPH3 links the Golgi, DNA damage, and cancer[J]. Cancer Res, 2015, 75(4): 624-627.
[13] Tokuda E, Itoh T, Hasegawa J, et al. Phosphatidylinositol 4-phosphate in the Golgi apparatus regulates cell-cell adhesion and invasive cell migration in human breast cancer[J]. Cancer Res, 2014, 74(11): 3054-3066.
[14]Chang SH, Hong SH, Jiang HL, et al. GOLGA2/GM130, cis-Golgi matrix protein, is a novel target of anticancer gene therapy[J]. Mol Ther, 2012, 20(11): 2052-2063.
[15]Fiore E, Vitti P. Serum TSH and risk of papillary thyroid cancer in nodular thyroid disease[J]. J Clin Endocrinol Metab, 2012, 97(4): 1134-1145.
[16]Romero N, Dumur CI, Martinez H, et al. Rab1b overexpression modifies Golgi size and gene expression in HeLa cells and modulates the thyrotrophin response in thyroid cells in culture[J] . Mol Biol Cell, 2013, 24(5): 617-632.
[17]Yuan H, Kajiyama H, Ito S, et al. ALX1 induces snail expression to promote epithelial-to-mesenchymal transition and invasion of ovarian cancer cells[J]. Cancer Res, 2013, 73(5): 1581-1590.
[18]Roy E, Bruyère J, Flamant P, et al. GM130 gain-of-function induces cell pathology in a model of lysosomal storage disease[J]. Hum Mol Genet, 2012, 21(7): 1481-1495.
[19]Denkert C, Budczies J, von Minckwitz G, et al. Strategies for developing Ki67 as a useful biomarker in breast cancer[J]. Breast, 2015, 24:S67-S72.
[20]Lingeman RG, Hickey RJ, Malkas LH. Expression of a novel peptide derived from PCNA damages DNA and reverses cisplatin resistance[J]. Cancer Chemother Pharmacol, 2014, 74(5): 981-993.
[21]Bhat G, Hothpet VR, Lin MF, et al. Shifted Golgi targeting of glycosyltransferases and α-mannosidase IA from giantin to GM130-GRASP65 results in formation of high mannose N-glycans in aggressive prostate cancer cells[J]. Biochim Biophys Acta Gen Subj, 2017, 1861(11 Pt A): 2891-2901.
[22]Shay G, Lynch CC, Fingleton B. Moving targets: Emerging roles for MMPs in Cancer Progression and Metastasis[J]. Matrix Biol, 2015, 44-46(3): 200-206.
[23]van Roy F. Beyond E-cadherin: roles of other cadherin superfamily members in cancer[J]. Nat Rev Cancer, 2014, 14(2): 121-134.
[24]Kidd ME, Shumaker DK, Ridge KM. The role of vimentin intermediate filaments in the progression of lung cancer[J]. Am J Respir Cell Mol Biol, 2014, 50(1): 1-6.
[25]Liang Z, Xie WJ, Zhao M, et al. DDR2 facilitates papillary thyroid carcinoma epithelial mesenchymal transition by activating ERK2/Snail1 pathway[J]. Oncol Lett, 2017, 14(6): 8114-8121.

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

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

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