Downregulated LRRK2 gene expression inhibits proliferation and migration while promoting the apoptosis of thyroid cancer cells by inhibiting activation of the JNK signaling pathway

Jiang,Zheng‑Cai; Chen,Xiao‑Jun; Zhou,Qi; Gong,Xiao‑Hua; Chen,Xiong; Wu,Wen‑Jun

doi:10.3892/ijo.2019.4816

Downregulated LRRK2 gene expression inhibits proliferation and migration while promoting the apoptosis of thyroid cancer cells by inhibiting activation of the JNK signaling pathway

Authors:
- Zheng‑Cai Jiang
- Xiao‑Jun Chen
- Qi Zhou
- Xiao‑Hua Gong
- Xiong Chen
- Wen‑Jun Wu
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Affiliations: Department of General Surgery, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China, Department of Endocrinology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325015, P.R. China
Published online on: May 29, 2019 https://doi.org/10.3892/ijo.2019.4816
Pages: 21-34
Copyright: © Jiang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Emerging studies have indicated that leucine‑rich repeat kinase 2 (LRRK2) is associated with thyroid cancer (TC). The present study investigated the effect of LRRK2 on the cell cycle and apoptosis in TC, and examined the underlying mechanisms in vitro. To screen TC‑associated differentially expressed genes, gene expression microarray analysis was conducted. Retrieval of pathways associated with TC from the Kyoto Encyclopedia of Genes and Genomes database indicated that the c‑Jun N‑terminal kinase (JNK) signaling pathway serves an essential role in TC. SW579, IHH‑4, TFC‑133, TPC‑1 and Nthy‑ori3‑1 cell lines were used to screen cell lines with the highest and lowest LRRK2 expression for subsequent experiments. The two selected cell lines were transfected with pcDNA‑LRRK2, or small interfering RNA against LRRK2 or SP600125 (a JNK inhibitor). Subsequently, flow cytometry, terminal deoxynucleotidyl transferase‑mediated dUTP‑biotin nick end labeling, a 5‑ethynyl‑2'‑deoxyuridine assay and a scratch test was conducted to detect the cell cycle distribution, apoptosis, proliferation and migration, respectively, in each group. The LRRK2 gene was determined to be elevated in TC based on the microarray data of the GSE3678 dataset. The SW579 cell line was identified to exhibit the highest LRRK2 expression, while IHH‑4 cells exhibited the lowest LRRK2 expression. LRRK2 silencing, through inhibiting the activation of the JNK signaling pathway, increased the expression levels of genes and proteins associated with cell cycle arrest and apoptosis in TC cells, promoted cell cycle arrest and apoptosis, and inhibited cell migration and proliferation in TC cells, indicating that LRRK2 repression could exert beneficial effects through the JNK signaling pathway on TC cells. These observations demonstrate that LRRK2 silencing promotes TC cell growth inhibition, and facilitates apoptosis and cell cycle arrest. The JNK signaling pathway may serve a crucial role in mediating the anti‑carcinogenic activities of downregulated LRRK2 in TC.

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1	Lun Y, Wu X, Xia Q, Han Y, Zhang X, Liu Z, Wang F, Duan Z, Xin S and Zhang J: Hashimoto's thyroiditis as a risk factor of papillary thyroid cancer may improve cancer prognosis. Otolaryngol Head Neck Surg. 148:396–402. 2013. View Article : Google Scholar : PubMed/NCBI
2	Nikiforov YE and Nikiforova MN: Molecular genetics and diagnosis of thyroid cancer. Nat Rev Endocrinol. 7:569–580. 2011. View Article : Google Scholar : PubMed/NCBI
3	Xing M: Molecular pathogenesis and mechanisms of thyroid cancer. Nat Rev Cancer. 13:184–199. 2013. View Article : Google Scholar : PubMed/NCBI
4	Nikiforov YE: Is ionizing radiation responsible for the increasing incidence of thyroid cancer? Cancer. 116:1626–1628. 2010. View Article : Google Scholar : PubMed/NCBI
5	Leux C, Truong T, Petit C, Baron-Dubourdieu D and Guénel P: Family history of malignant and benign thyroid diseases and risk of thyroid cancer: A population-based case-control study in New Caledonia. Cancer Causes Control. 23:745–755. 2012. View Article : Google Scholar : PubMed/NCBI
6	Brindel P, Doyon F, Bourgain C, Rachédi F, Boissin JL, Sebbag J, Shan L, Bost-Bezeaud F, Petitdidier P, Paoaafaite J, et al: Family history of thyroid cancer and the risk of differentiated thyroid cancer in French polynesia. Thyroid. 20:393–400. 2010. View Article : Google Scholar : PubMed/NCBI
7	Pellegriti G, Frasca F, Regalbuto C, Squatrito S and Vigneri R: Worldwide increasing incidence of thyroid cancer: Update on epidemiology and risk factors. J Cancer Epidemiol. 2013:9652122013. View Article : Google Scholar : PubMed/NCBI
8	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
9	Davies L and Welch HG: Current thyroid cancer trends in the United States. JAMA Otolaryngol Head Neck Surg. 140:317–322. 2014. View Article : Google Scholar : PubMed/NCBI
10	Nexø MA, Watt T, Cleal B, Hegedüs L, Bonnema SJ, Rasmussen AK, Feldt-Rasmussen U and Bjorner JB: Exploring the experiences of people with hypo- and hyperthyroidism. Qual Health Res. 25:945–953. 2015. View Article : Google Scholar
11	Rowe CW, Bendinelli C and McGrath S: Charting a course through the CEAs: Diagnosis and management of medullary thyroid cancer. Clin Endocrinol (Oxf). 85:340–343. 2016. View Article : Google Scholar
12	Lubitz CC and Sosa JA: The changing landscape of papillary thyroid cancer: Epidemiology, management, and the implications for patients. Cancer. 122:3754–3759. 2016. View Article : Google Scholar : PubMed/NCBI
13	Al-Humadi H, Zarros A, Al-Saigh R and Liapi C: Genetic basis and gene therapy trials for thyroid cancer. Cancer Genomics Proteomics. 7:31–49. 2010.PubMed/NCBI
14	Nikonova EV, Xiong Y, Tanis KQ, Dawson VL, Vogel RL, Finney EM, Stone DJ, Reynolds IJ, Kern JT and Dawson TM: Transcriptional responses to loss or gain of function of the leucine-rich repeat kinase 2 (LRRK2) gene uncover biological processes modulated by LRRK2 activity. Hum Mol Genet. 21. pp. 163–174. 2012, View Article : Google Scholar
15	Kubo M, Kamiya Y, Nagashima R, Maekawa T, Eshima K, Azuma S, Ohta E and Obata F: LRRK2 is expressed in B-2 but not in B-1 B cells, and downregulated by cellular activation. J Neuroimmunol. 229:123–128. 2010. View Article : Google Scholar : PubMed/NCBI
16	Maekawa T, Mori S, Sasaki Y, Miyajima T, Azuma S, Ohta E and Obata F: The I2020T Leucine-rich repeat kinase 2 transgenic mouse exhibits impaired locomotive ability accompanied by dopaminergic neuron abnormalities. Mol Neurodegener. 7:152012. View Article : Google Scholar : PubMed/NCBI
17	Piccoli G, Condliffe SB, Bauer M, Giesert F, Boldt K, De Astis S, Meixner A, Sarioglu H, Vogt-Weisenhorn DM, Wurst W, et al: LRRK2 controls synaptic vesicle storage and mobilization within the recycling pool. J Neurosci. 31:2225–2237. 2011. View Article : Google Scholar : PubMed/NCBI
18	Zechel S, Meinhardt A, Unsicker K and von Bohlen Und Halbach O: Expression of leucine-rich-repeat-kinase 2 (LRRK2) during embryonic development. Int J Dev Neurosci. 28:391–399. 2010. View Article : Google Scholar : PubMed/NCBI
19	Looyenga BD, Furge KA, Dykema KJ, Koeman J, Swiatek PJ, Giordano TJ, West AB, Resau JH, Teh BT and MacKeigan JP: Chromosomal amplification of leucine-rich repeat kinase-2 (LRRK2) is required for oncogenic MET signaling in papillary renal and thyroid carcinomas. Proc Natl Acad Sci USA. 108:1439–1444. 2011. View Article : Google Scholar : PubMed/NCBI
20	Wang LL, Huang H, Zhang CR, Xia J, Liu SS and Wang XW: Cloning and functional characterization of c-Jun NH2-terminal kinase from the Mediterranean species of the Whitefly Bemisia tabaci complex. Int J Mol Sci. 14:13433–13446. 2013. View Article : Google Scholar : PubMed/NCBI
21	Xu B, Yang H, Sun M, Chen H, Jiang L, Zheng X, Ding G, Liu Y, Sheng Y, Cui D, et al: 2,3′,4,4′,5-pentachlorobiphenyl induces inflammatory responses in the thyroid through JNK and Aryl hydrocarbon receptor-mediated pathway. Toxicol Sci. 149:300–311. 2016. View Article : Google Scholar
22	Shang J, Ding Q, Yuan S, Liu JX, Li F and Zhang H: Network analyses of integrated differentially expressed genes in papillary thyroid carcinoma to identify characteristic genes. Genes (Basel). 10. pp. E452019, View Article : Google Scholar
23	Szklarczyk D, Morris JH, Cook H, Kuhn M, Wyder S, Simonovic M, Santos A, Doncheva NT, Roth A, Bork P, et al: The STRING database in 2017: Quality-controlled protein-protein association networks, made broadly accessible. Nucleic Acids Res. 45(D1): D362–D368. 2017. View Article : Google Scholar
24	Piñero J, Bravo À, Queralt-Rosinach N, Gutiérrez-Sacristán A, Deu-Pons J, Centeno E, García-García J, Sanz F and Furlong LI; DisGeNET: A comprehensive platform integrating information on human disease-associated genes and variants. Nucleic Acids Res. 45(D1): D833–D839. 2017. View Article : Google Scholar
25	Piñero J, Queralt-Rosinach N, Bravo À, Deu-Pons J, Bauer-Mehren A, Baron M, Sanz F and Furlong LI; DisGeNET: A discovery platform for the dynamical exploration of human diseases and their genes. Database (Oxford). 2015. pp. bav0282015, View Article : Google Scholar
26	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-∆ ∆ C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
27	Santos LS, Branco SC, Silva SN, Azevedo AP, Gil OM, Manita I, Ferreira TC, Limbert E, Rueff J and Gaspar JF: Polymorphisms in base excision repair genes and thyroid cancer risk. Oncol Rep. 28:1859–1868. 2012. View Article : Google Scholar : PubMed/NCBI
28	Stjepanovic N and Capdevila J: Multikinase inhibitors in the treatment of thyroid cancer: Specific role of lenvatinib. Biologics. 8:129–139. 2014.PubMed/NCBI
29	Maekawa T, Kubo M, Yokoyama I, Ohta E and Obata F: Age-dependent and cell-population-restricted LRRK2 expression in normal mouse spleen. Biochem Biophys Res Commun. 392:431–435. 2010. View Article : Google Scholar : PubMed/NCBI
30	Sabapathy K: Role of the JNK pathway in human diseases. Prog Mol Biol Transl Sci. 106:145–169. 2012. View Article : Google Scholar : PubMed/NCBI
31	Dou J, Li X, Cai Y, Chen H, Zhu S, Wang Q, Zou X, Mei Y, Yang Q, Li W, et al: Human cytomegalovirus induces caspase-dependent apoptosis of megakaryocytic CHRF-288-11 cells by activating the JNK pathway. Int J Hematol. 91:620–629. 2010. View Article : Google Scholar : PubMed/NCBI
32	Wang X, Chao L, Li X, Ma G, Chen L, Zang Y and Zhou G: Elevated expression of phosphorylated c-Jun NH2-terminal kinase in basal-like and 'triple-negative' breast cancers. Hum Pathol. 41:401–406. 2010. View Article : Google Scholar
33	Trancikova A, Mamais A, Webber PJ, Stafa K, Tsika E, Glauser L, West AB, Bandopadhyay R and Moore DJ: Phosphorylation of 4E-BP1 in the mammalian brain is not altered by LRRK2 expression or pathogenic mutations. PLoS One. 7:e477842012. View Article : Google Scholar : PubMed/NCBI
34	Kwon SH, Kim JA, Hong SI, Jung YH, Kim HC, Lee SY and Jang CG: Loganin protects against hydrogen peroxide-induced apoptosis by inhibiting phosphorylation of JNK, p38, and ERK 1/2 MAPKs in SH-SY5Y cells. Neurochem Int. 58:533–541. 2011. View Article : Google Scholar : PubMed/NCBI
35	Kim MK, Choi HS, Cho SG, Shin YC and Ko SG: Rubus coreanus Miquel extract causes apoptosis of doxorubicin-resistant NCI/ADR-RES ovarian cancer cells via JNK phosphorylation. Mol Med Rep. 13:4065–4072. 2016. View Article : Google Scholar : PubMed/NCBI
36	Wang L, Tian Z, Yang Q, Li H, Guan H, Shi B, Hou P and Ji M: Sulforaphane inhibits thyroid cancer cell growth and invasiveness through the reactive oxygen species-dependent pathway. Oncotarget. 6:25917–25931. 2015.PubMed/NCBI
37	Guan H, Liang W, Liu J, Wei G, Li H, Xiu L, Xiao H and Li Y: Transmembrane protease serine 4 promotes thyroid cancer proliferation via CREB phosphorylation. Thyroid. 25:85–94. 2015. View Article : Google Scholar :
38	Lu Y, Wei C and Xi Z: Curcumin suppresses proliferation and invasion in non-small cell lung cancer by modulation of MTA1-mediated Wnt/β-catenin pathway. In Vitro Cell Dev Biol Anim. 50:840–850. 2014. View Article : Google Scholar : PubMed/NCBI
39	Sui C, Zhuang C, Sun D, Yang L, Zhang L and Song L: Notch1 regulates the JNK signaling pathway and increases apoptosis in hepatocellular carcinoma. Oncotarget. 8:45837–45847. 2017. View Article : Google Scholar : PubMed/NCBI
40	Husdal A, Bukholm G and Bukholm IR: The prognostic value and overexpression of cyclin A is correlated with gene amplification of both cyclin A and cyclin E in breast cancer patient. Cell Oncol. 28:107–116. 2006.PubMed/NCBI
41	Romagosa C, Simonetti S, López-Vicente L, Mazo A, Lleonart ME, Castellvi J and Ramon y Cajal S: p16(Ink4a) over-expression in cancer: A tumor suppressor gene associated with senescence and high-grade tumors. Oncogene. 30:2087–2097. 2011. View Article : Google Scholar : PubMed/NCBI
42	Li Y, Shen L, Xu H, Pang Y, Xu Y, Ling M, Zhou J, Wang X and Liu Q: Up-regulation of cyclin D1 by JNK1/c-Jun is involved in tumorigenesis of human embryo lung fibroblast cells induced by a low concentration of arsenite. Toxicol Lett. 206:113–120. 2011. View Article : Google Scholar : PubMed/NCBI
43	Poomsawat S, Buajeeb W, Khovidhunkit SO and Punyasingh J: Alteration in the expression of cdk4 and cdk6 proteins in oral cancer and premalignant lesions. J Oral Pathol Med. 39:793–799. 2010. View Article : Google Scholar : PubMed/NCBI
44	Schapansky J, Nardozzi JD and LaVoie MJ: The complex relationships between microglia, alpha-synuclein, and LRRK2 in Parkinson's disease. Neuroscience. 302:74–88. 2015. View Article : Google Scholar
45	Berwick DC and Harvey K: LRRK2 signaling pathways: The key to unlocking neurodegeneration? Trends Cell Biol. 21:257–265. 2011. View Article : Google Scholar : PubMed/NCBI
46	Chen CY, Weng YH, Chien KY, Lin KJ, Yeh TH, Cheng YP, Lu CS and Wang HL: (G2019S) LRRK2 activates MKK4-JNK pathway and causes degeneration of SN dopaminergic neurons in a transgenic mouse model of PD. Cell Death Differ. 19:1623–1633. 2012. View Article : Google Scholar : PubMed/NCBI
47	Luerman GC, Nguyen C, Samaroo H, Loos P, Xi H, Hurtado-Lorenzo A, Needle E, Stephen Noell G, Galatsis P, Dunlop J, et al: Phosphoproteomic evaluation of pharmacological inhibition of leucine-rich repeat kinase 2 reveals significant off-target effects of LRRK-2-IN-1. J Neurochem. 128:561–576. 2014. View Article : Google Scholar
48	Moehle MS, Webber PJ, Tse T, Sukar N, Standaert DG, DeSilva TM, Cowell RM and West AB: LRRK2 inhibition attenuates microglial inflammatory responses. J Neurosci. 32:1602–1611. 2012. View Article : Google Scholar : PubMed/NCBI
49	Prabhudesai S, Bensabeur FZ, Abdullah R, Basak I, Baez S, Alves G, Holtzman NG, Larsen JP and Møller SG: LRRK2 knockdown in zebrafish causes developmental defects, neuronal loss, and synuclein aggregation. J Neurosci Res. 94:717–735. 2016. View Article : Google Scholar : PubMed/NCBI
50	Xu P, Xia X, Yang Z, Tian Y, Di J and Guo M: Silencing of TCTN1 inhibits proliferation, induces cell cycle arrest and apoptosis in human thyroid cancer. Exp Ther Med. 14:3720–3726. 2017. View Article : Google Scholar : PubMed/NCBI