Knockdown of ARK5 expression suppresses invasion of ovarian cancer cells

  • Authors:
    • Shuxiao Wang
    • Shuwei Li
    • Hui Wang
    • Wei Li
    • Yuxue Gao
    • Xuejian Wang
    • Chunyan Fang
    • Baogang Zhang
    • Xiuning Sun
    • Ruifang Li
    • Weiwei Shi
    • Meiling Chen
    • Lihong Shi
  • View Affiliations

  • Published online on: January 28, 2019     https://doi.org/10.3892/mmr.2019.9901
  • Pages: 2927-2934
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The aim of the current study was to investigate the effects and the molecular mechanisms of ARK5 in ovarian cancer cell invasion. The plasmid pGCsilencerU6/GFP/Neo‑RNAi‑ARK5 and the control vector with a scramble sequence were transfected into SKOV3 cells to establish ARK5‑deficient SKOV3 cells (siARK5/SKOV3) and a control cell line (Scr/SKOV3), respectively. Reverse transcription‑polymerase chain reaction (RT‑PCR) and Western blot analysis were used to determine the mRNA and protein expression levels of ARK5. Migration and invasion abilities of SKOV3 cells were determined in chemotaxis and invasion assays, respectively. The epidermal growth factor‑1 (EGF‑1)‑induced expression of matrix metallopeptidase (MMP)‑2 and MMP‑9, epithelial‑mesenchymal transition (EMT) and phosphorylation of mechanistic target of rapamycin kinase (mTOR) in siARK5/SKOV3 and Scr/SKOV3 cells were detected by western blot. RT‑PCR and western blot analyses demonstrated that the expression of ARK5 was significantly downregulated in siARK5/SKOV3 cells at the mRNA and protein levels (P<0.01). The migration and invasion abilities of siARK5/SKOV3 cells were markedly decreased compared with Scr/SKOV3 cells (P<0.01). In addition, the results demonstrated that EGF‑1‑induced expression of MMP‑2 and MMP‑9, EMT and phosphorylation of mTOR were suppressed in siARK5/SKOV3 cells as compared with Scr/SKOV3 cells (P<0.01). The current study demonstrated that ARK5 is a critical factor involved in SKOV3 cell invasion and ARK5 increases invasive potential by promoting EMT and activating the Akt‑mTOR‑MMPs pathway.

References

1 

Bowtell DD: The genesis and evolution of high-grade serous ovarian cancer. Nat Rev Cancer. 10:803–808. 2010. View Article : Google Scholar : PubMed/NCBI

2 

Siegel RL, Miller KD, Fedewa SA, Ahnen DJ, Meester RGS, Barzi A and Jemal A: Colorectal cancer statistics, 2017. CA Cancer J Clin. 67:177–193. 2017. View Article : Google Scholar : PubMed/NCBI

3 

Ledermann JA, Embleton AC, Raja F, Perren TJ, Jayson GC, Rustin GJS, Kaye SB, Hirte H, Eisenhauer E, Vaughan M, et al: Cediranib in patients with relapsed platinum-sensitive ovarian cancer (ICON6): A randomised, double-blind, placebo-controlled phase 3 trial. Lancet. 387:1066–1074. 2016. View Article : Google Scholar : PubMed/NCBI

4 

Shevade A, Strogolova V, Orlova M, Yeo CT and Kuchin S: Mitochondrial voltage-dependent anion channel protein Por1 positively regulates the nuclear localization of saccharomyces cerevisiae AMP-activated protein kinase. mSphere. 3(pii): e00482–17. 2018.PubMed/NCBI

5 

Zhang X, Lv H, Zhou Q, Elkholi R, Chipuk JE, Reddy MV, Reddy EP and Gallo JM: Preclinical pharmacological evaluation of a novel multiple kinase inhibitor, ON123300, in brain tumor models. Mol Cancer Ther. 13:1105–1116. 2014. View Article : Google Scholar : PubMed/NCBI

6 

Cao D, Li D, Huang Y, Ma Y, Zhang B, Zhao C, Deng S, Luo M, Yin T, Wei YQ and Wang W: 5-azacytidine promotes invadopodia formation and tumor metastasis through the upregulation of PI3K in ovarian cancer cells. Oncotarget. 8:60173–60187. 2017.PubMed/NCBI

7 

Nehate C, Moothedathu Raynold AA and Koul V: ATRP fabricated and short chain polyethylenimine grafted redox sensitive polymeric nanoparticles for codelivery of anticancer drug and siRNA in cancer therapy. ACS Appl Mater Interfaces. 9:39672–39687. 2017. View Article : Google Scholar : PubMed/NCBI

8 

Stubelius A, Andersson A, Islander U and Carlsten H: Ovarian hormones in innate inflammation. Immunobiology. 222:878–883. 2017. View Article : Google Scholar : PubMed/NCBI

9 

Li L, Gong M, Zhao Y, Zhao X and Li Q: FOXK1 facilitates cell proliferation through regulating the expression of p21, and promotes metastasis in ovarian cancer. Oncotarget. 8:70441–70451. 2017.PubMed/NCBI

10 

Peng F, Zhong Y, Liu Y, Zhang Y, Xie Y, Lu Y, Zhang X and Li D: SPARC suppresses lymph node metastasis by regulating the expression of VEGFs in ovarian carcinoma. Int J Oncol. 51:1920–1928. 2017. View Article : Google Scholar : PubMed/NCBI

11 

Mu QJ, Li HL, Yao Y, Liu SC, Yin CG and Ma XZ: Chromodomain helicase/ATPase DNA-binding protein 1-like gene (CHD1L) expression and implications for invasion and metastasis of breast cancer. PLoS One. 10:e01430302015. View Article : Google Scholar : PubMed/NCBI

12 

Suzuki A, Lu J, Kusakai G, Kishimoto A, Ogura T and Esumi H: ARK5 is a tumor invasion-associated factor downstream of Akt signaling. Mol Cell Biol. 24:3526–3535. 2004. View Article : Google Scholar : PubMed/NCBI

13 

Shi L, Sun X, Zhang J, Zhao C, Li H, Liu Z, Fang C, Wang X, Zhao C, Zhang X, et al: Gab2 expression in glioma and its implications for tumor invasion. Acta Oncol. 52:1739–1750. 2013. View Article : Google Scholar : PubMed/NCBI

14 

Tang J, Qin Z, Han P, Wang W, Yang C, Xu Z, Li R, Liu B, Qin C, Wang Z, et al: High Annexin A5 expression promotes tumor progression and poor prognosis in renal cell carcinoma. Int J Oncol. 50:1839–1847. 2017. View Article : Google Scholar : PubMed/NCBI

15 

Wang Y and Zhou BP: Epithelial-mesenchymal transition in breast cancer progression and metastasis. Chin J Cancer. 30:603–611. 2011. View Article : Google Scholar : PubMed/NCBI

16 

Xu T, Zhang J, Chen W, Pan S, Zhi X, Wen L, Zhou Y, Chen BW, Qiu J, Zhang Y, et al: ARK5 promotes doxorubicin resistance in hepatocellular carcinoma via epithelial-mesenchymal transition. Cancer Lett. 377:140–148. 2016. View Article : Google Scholar : PubMed/NCBI

17 

Lue H, Thiele M, Franz J, Dahl E, Speckgens S, Leng L, Fingerle-Rowson G, Bucala R, Lüscher B and Bernhagen J: Macrophage migration inhibitory factor (MIF) promotes cell survival by activation of the Akt pathway and role for CSN5/JAB1 in the control of autocrine MIF activity. Oncogene. 26:5046–5059. 2007. View Article : Google Scholar : PubMed/NCBI

18 

Lamouille S, Connolly E, Smyth JW, Akhurst RJ and Derynck R: TGF-β-induced activation of mTOR complex 2 drives epithelial-mesenchymal transition and cell invasion. J Cell Sci. 125:1259–1273. 2012. View Article : Google Scholar : PubMed/NCBI

19 

Li M, Zheng C, Xu H, He W, Ruan Y, Ma J, Zheng J, Ye C and Li W: Inhibition of AMPK-related kinase 5 (ARK5) enhances cisplatin cytotoxicity in non-small cell lung cancer cells through regulation of epithelial-mesenchymal transition. Am J Transl Res. 9:1708–1719. 2017.PubMed/NCBI

20 

Kusakai G, Suzuki A, Ogura T, Kaminishi M and Esumi H: Strong association of ARK5 with tumor invasion and metastasis. J Exp Clin Cancer Res. 23:263–268. 2004.PubMed/NCBI

21 

Lu S, Niu N, Guo H, Tang J, Guo W, Liu Z, Shi L, Sun T, Zhou F, Li H, et al: ARK5 promotes glioma cell invasion, and its elevated expression is correlated with poor clinical outcome. Eur J Cancer. 49:752–763. 2013. View Article : Google Scholar : PubMed/NCBI

22 

Zhang HY, Li JH, Li G and Wang SR: Activation of ARK5/miR-1181/HOXA10 axis promotes epithelial-mesenchymal transition in ovarian cancer. Oncol Rep. 34:1193–1202. 2015. View Article : Google Scholar : PubMed/NCBI

23 

Phippen NT, Bateman NW, Wang G, Conrads KA, Ao W, Teng PN, Litzi TA, Oliver J, Maxwell GL, Hamilton CA, et al: NUAK1 (ARK5) is associated with poor prognosis in ovarian cancer. Front Oncol. 6:2132016. View Article : Google Scholar : PubMed/NCBI

24 

Roh SA, Choi EY, Cho DH, Jang SJ, Kim SY, Kim YS and Kim JC: Growth and invasion of sporadic colorectal adenocarcinomas in terms of genetic change. J Korean Med Sci. 25:353–360. 2010. View Article : Google Scholar : PubMed/NCBI

25 

Long H, Xie R, Xiang T, Zhao Z, Lin S, Liang Z, Chen Z and Zhu B: Autocrine CCL5 signaling promotes invasion and migration of CD133+ ovarian cancer stem-like cells via NF-κB-mediated MMP-9 upregulation. Stem Cells. 30:2309–2319. 2012. View Article : Google Scholar : PubMed/NCBI

26 

Liao TT and Yang MH: Revisiting epithelial-mesenchymal transition in cancer metastasis: The connection between epithelial plasticity and stemness. Mol Oncol. 11:792–804. 2017. View Article : Google Scholar : PubMed/NCBI

27 

Shi L, Zhang B, Sun X, Lu S, Liu Z, Liu Y, Li H, Wang L, Wang X and Zhao C: MiR-204 inhibits human NSCLC metastasis through suppression of NUAK1. Br J Cancer. 111:2316–2327. 2014. View Article : Google Scholar : PubMed/NCBI

28 

Suzuki A, Kusakai G, Kishimoto A, Lu J, Ogura T, Lavin MF and Esumi H: Identification of a novel protein kinase mediating Akt survival signaling to the ATM protein. J Biol Chem. 278:48–53. 2003. View Article : Google Scholar : PubMed/NCBI

29 

Yu J, Tao S, Hu P, Wang R, Fang C, Xu Y, Qi D, Wei Z, Zhang J and Tan Q: CCR7 promote lymph node metastasis via regulating VEGF-C/D-R3 pathway in lung adenocarcinoma. J Cancer. 8:2060–2068. 2017. View Article : Google Scholar : PubMed/NCBI

30 

Martin TA and Jiang WG: Anti-cancer agents in medicinal chemistry (formerly current medicinal chemistry-anti-cancer agents). Anticancer Agents Med Chem. 10:12010. View Article : Google Scholar : PubMed/NCBI

31 

Cheng K and Hao M: Metformin inhibits TGF-β1-induced epithelial-to-mesenchymal transition via PKM2 relative-mTOR/p70s6k signaling pathway in cervical carcinoma cells. Int J Mol Sci. 17(pii): E20002016. View Article : Google Scholar : PubMed/NCBI

32 

Suzuki A, Kusakai G, Kishimoto A, Shimojo Y, Miyamoto S, Ogura T, Ochiai A and Esumi H: Regulation of caspase-6 and FLIP by the AMPK family member ARK5. Oncogene. 23:7067–7075. 2004. View Article : Google Scholar : PubMed/NCBI

33 

Brunet A, Bonni A, Zigmond MJ, Lin MZ, Juo P, Hu LS, Anderson MJ, Arden KC, Blenis J and Greenberg ME: Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell. 96:857–868. 1999. View Article : Google Scholar : PubMed/NCBI

34 

Chen D, Liu G, Xu N, You X, Zhou H, Zhao X and Liu Q: Knockdown of ARK5 expression suppresses invasion and metastasis of gastric cancer. Cell Physiol Biochem. 42:1025–1036. 2017. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

April 2019
Volume 19 Issue 4

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Wang, S., Li, S., Wang, H., Li, W., Gao, Y., Wang, X. ... Shi, L. (2019). Knockdown of ARK5 expression suppresses invasion of ovarian cancer cells. Molecular Medicine Reports, 19, 2927-2934. https://doi.org/10.3892/mmr.2019.9901
MLA
Wang, S., Li, S., Wang, H., Li, W., Gao, Y., Wang, X., Fang, C., Zhang, B., Sun, X., Li, R., Shi, W., Chen, M., Shi, L."Knockdown of ARK5 expression suppresses invasion of ovarian cancer cells". Molecular Medicine Reports 19.4 (2019): 2927-2934.
Chicago
Wang, S., Li, S., Wang, H., Li, W., Gao, Y., Wang, X., Fang, C., Zhang, B., Sun, X., Li, R., Shi, W., Chen, M., Shi, L."Knockdown of ARK5 expression suppresses invasion of ovarian cancer cells". Molecular Medicine Reports 19, no. 4 (2019): 2927-2934. https://doi.org/10.3892/mmr.2019.9901