Thymoquinone inhibits metastatic phenotype and epithelial‑mesenchymal transition in renal cell carcinoma by regulating the LKB1/AMPK signaling pathway

Kou,Bo; Kou,Qingshan; Ma,Bo; Zhang,Jing; Sun,Borui; Yang,Yang; Li,Jianpeng; Zhou,Jinsong; Liu,Wei

doi:10.3892/or.2018.6519

Thymoquinone inhibits metastatic phenotype and epithelial‑mesenchymal transition in renal cell carcinoma by regulating the LKB1/AMPK signaling pathway

Authors:
- Bo Kou
- Qingshan Kou
- Bo Ma
- Jing Zhang
- Borui Sun
- Yang Yang
- Jianpeng Li
- Jinsong Zhou
- Wei Liu
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Affiliations: Department of Cardiovascular Surgery, The First Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Medical Center, The First People's Hospital of Xianyang, Xianyang, Shaanxi 712000, P.R. China, Department of Ophthalmology, The First Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Gynaecology, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710061, P.R. China, Department of Anesthesiology, The First Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Department of Human Anatomy, Histology and Embryology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi 710061, P.R. China, Department of Vascular Surgery, The First Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China
Published online on: June 25, 2018 https://doi.org/10.3892/or.2018.6519
Pages: 1443-1450

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Abstract

Thymoquinone, isolated from the seeds of Nigella sativa, has exhibited antitumor properties in a variety of cancer types. However, few studies have investigated the effect of thymoquinone (TQ) on migration and invasion in renal cell carcinoma (RCC). In the present study, our results confirmed that TQ significantly inhibited the migration and invasion of the human RCC 769‑P and 786‑O cell lines, as demonstrated by wound healing and Transwell assays. Additionally, TQ upregulated the expression of E‑cadherin and downregulated the expression of Snail, ZEB1 and vimentin at the mRNA and protein levels in a concentration‑dependent manner. Subsequently, the phosphorylation levels of liver kinase B1 (LKB1) and AMP‑activated protein kinase (AMPK) were increased upon TQ treatment. To further validate the role of LKB1/AMPK signaling, we revealed that TQ‑mediated increase of E‑cadherin level and reduction of Snail level could be further enhanced by LKB1 overexpression. Furthermore, co‑treatment with the AMPK inhibitor Compound C attenuated the anti‑metastatic effect of TQ on RCC and partially abrogated the high expression of E‑cadherin and the low expression of Snail mediated by TQ. In contrast, the AMPK activator AICAR demonstrated the opposite effect. Collectively, the present study revealed that TQ could markedly suppress the metastatic phenotype and reverse the epithelial‑mesenchymal transition in RCC by regulating the LKB1/AMPK signaling pathway, indicating that TQ may be a potential therapeutic candidate against RCC.

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1	Taniguchi H, Ito S, Ueda T, Morioka Y, Kayukawa N, Ueno A, Nakagawa H, Fujihara A, Ushijima S, Kanazawa M, et al: CNPY2 promoted the proliferation of renal cell carcinoma cells and increased the expression of TP53. Biochem Biophys Res Commun. 485:267–271. 2017. View Article : Google Scholar : PubMed/NCBI
2	Hirata H, Hinoda Y, Nakajima K, Kawamoto K, Kikuno N, Ueno K, Yamamura S, Zaman MS, Khatri G, Chen Y, et al: Wnt antagonist DKK1 acts as a tumor suppressor gene that induces apoptosis and inhibits proliferation in human renal cell carcinoma. Int J Cancer. 128:1793–1803. 2011. View Article : Google Scholar : PubMed/NCBI
3	Hsu HH, Chen MC, Day CH, Lin YM, Li SY, Tu CC, Padma VV, Shih HN, Kuo WW and Huang CY: Thymoquinone suppresses migration of LoVo human colon cancer cells by reducing prostaglandin E2 induced COX-2 activation. World J Gastroenterol. 23:1171–1179. 2017. View Article : Google Scholar : PubMed/NCBI
4	Hossen MJ, Yang WS, Kim D, Aravinthan A, Kim JH and Cho JY: Thymoquinone: An IRAK1 inhibitor with in vivo and in vitro anti-inflammatory activities. Sci Rep. 7:429952017. View Article : Google Scholar : PubMed/NCBI
5	Cobourne-Duval MK, Taka E, Mendonca P, Bauer D and Soliman KF: The antioxidant effects of thymoquinone in activated BV-2 murine microglial cells. Neurochem Res. 41:3227–3238. 2016. View Article : Google Scholar : PubMed/NCBI
6	Ke X, Zhao Y, Lu X, Wang Z, Liu Y, Ren M, Lu G, Zhang D, Sun Z, Xu Z, et al: TQ inhibits hepatocellular carcinoma growth in vitro and in vivo via repression of Notch signaling. Oncotarget. 6:32610–32621. 2015. View Article : Google Scholar : PubMed/NCBI
7	Taha MM, Sheikh BY, Salim LZ, Mohan S, Khan A, Kamalidehghan B, Ahmadipour F and Abdelwahab SI: Thymoquinone induces apoptosis and increase ROS in ovarian cancer cell line. Cell Mol Biol (Noisy-le-grand). 62:97–101. 2016.PubMed/NCBI
8	Yang J, Kuang XR, Lv PT and Yan XX: Thymoquinone inhibits proliferation and invasion of human nonsmall-cell lung cancer cells via ERK pathway. Tumour Biol. 36:259–269. 2015. View Article : Google Scholar : PubMed/NCBI
9	Wilson AJ, Saskowski J, Barham W, Yull F and Khabele D: Thymoquinone enhances cisplatin-response through direct tumor effects in a syngeneic mouse model of ovarian cancer. J Ovarian Res. 8:462015. View Article : Google Scholar : PubMed/NCBI
10	Park EJ, Chauhan AK, Min KJ, Park DC and Kwon TK: Thymoquinone induces apoptosis through downregulation of c-FLIP and Bcl-2 in renal carcinoma Caki cells. Oncol Rep. 36:2261–2267. 2016. View Article : Google Scholar : PubMed/NCBI
11	Jeghers H, Mckusick VA and Katz KH: Generalized intestinal polyposis and melanin spots of the oral mucosa, lips and digits; a syndrome of diagnostic significance. N Engl J Med. 241:1031–1036. 1949. View Article : Google Scholar : PubMed/NCBI
12	LKB1 and AMPK maintain epithelial cell polarity under energetic stress. J Cell Biol. 203:3732013. View Article : Google Scholar : PubMed/NCBI
13	Dahmani R, Just PA, Delay A, Canal F, Finzi L, Prip-Buus C, Lambert M, Sujobert P, Buchet-Poyau K, Miller E, et al: A novel LKB1 isoform enhances AMPK metabolic activity and displays oncogenic properties. Oncogene. 34:2337–2346. 2015. View Article : Google Scholar : PubMed/NCBI
14	Kan JY, Yen MC, Wang JY, Wu DC, Chiu YJ, Ho YW and Kuo PL: Nesfatin-1/Nucleobindin-2 enhances cell migration, invasion, and epithelial-mesenchymal transition via LKB1/AMPK/TORC1/ZEB1 pathways in colon cancer. Oncotarget. 7:31336–31349. 2016. View Article : Google Scholar : PubMed/NCBI
15	Taliaferro-Smith L, Nagalingam A, Zhong D, Zhou W, Saxena NK and Sharma D: LKB1 is required for adiponectin-mediated modulation of AMPK-S6K axis and inhibition of migration and invasion of breast cancer cells. Oncogene. 28:2621–2633. 2009. View Article : Google Scholar : PubMed/NCBI
16	Marcus AI and Zhou W: LKB1 regulated pathways in lung cancer invasion and metastasis. J Thorac Oncol. 5:1883–1886. 2010. View Article : Google Scholar : PubMed/NCBI
17	Li X, Teng S, Zhang Y, Zhang W, Zhang X, Xu K, Yao H, Yao J, Wang H, Liang X and Hu Z: TROP2 promotes proliferation, migration and metastasis of gallbladder cancer cells by regulating PI3K/AKT pathway and inducing EMT. Oncotarget. 8:47052–47063. 2017.PubMed/NCBI
18	Li N, Huang D, Lu N and Luo L: Role of the LKB1/AMPK pathway in tumor invasion and metastasis of cancer cells (Review). Oncol Rep. 34:2821–2826. 2015. View Article : Google Scholar : PubMed/NCBI
19	Ahmad I, Muneer KM, Tamimi IA, Chang ME, Ata MO and Yusuf N: Thymoquinone suppresses metastasis of melanoma cells by inhibition of NLRP3 inflammasome. Toxicol Appl Pharmacol. 270:70–76. 2013. View Article : Google Scholar : PubMed/NCBI
20	Kolli-Bouhafs K, Boukhari A, Abusnina A, Velot E, Gies JP, Lugnier C and Rondé P: Thymoquinone reduces migration and invasion of human glioblastoma cells associated with FAK, MMP-2 and MMP-9 down-regulation. Invest New Drugs. 30:2121–2131. 2012. View Article : Google Scholar : PubMed/NCBI
21	Zhou Y, Zhu Y, Fan X, Zhang C, Wang Y, Zhang L, Zhang H, Wen T, Zhang K, Huo X, et al: NID1, a new regulator of EMT required for metastasis and chemoresistance of ovarian cancer cells. Oncotarget. 8:33110–33121. 2017.PubMed/NCBI
22	Zhao S, Sun H, Jiang W, Mi Y, Zhang D, Wen Y, Cheng D, Tang H, Wu S, Yu Y, et al: miR-4775 promotes colorectal cancer invasion and metastasis via the Smad7/TGFβ-mediated epithelial to mesenchymal transition. Mol Cancer. 16:122017. View Article : Google Scholar : PubMed/NCBI
23	Khan MA, Tania M, Wei C, Mei Z, Fu S, Cheng J, Xu J and Fu J: Thymoquinone inhibits cancer metastasis by downregulating TWIST1 expression to reduce epithelial to mesenchymal transition. Oncotarget. 6:19580–19591. 2015. View Article : Google Scholar : PubMed/NCBI
24	Weng J, Zhang H, Wang C, Liang J, Chen G, Li W, Tang H and Hou J: miR-373-3p targets DKK1 to promote EMT-induced metastasis via the Wnt/β-catenin pathway in tongue squamous cell carcinoma. Biomed Res Int. 2017:60109262017. View Article : Google Scholar : PubMed/NCBI
25	Chen J, Zhang H, Chen Y, Qiao G, Jiang W, Ni P, Liu X and Ma L: miR-598 inhibits metastasis in colorectal cancer by suppressing JAG1/Notch2 pathway stimulating EMT. Exp Cell Res. 352:104–112. 2017. View Article : Google Scholar : PubMed/NCBI
26	Wei S, Wang L, Zhang L, Li B, Li Z, Zhang Q, Wang J, Chen L, Sun G, Li Q, et al: ZNF143 enhances metastasis of gastric cancer by promoting the process of EMT through PI3K/AKT signaling pathway. Tumour Biol. 37:12813–12821. 2016. View Article : Google Scholar : PubMed/NCBI
27	Li J, Liu J, Li P, Mao X, Li W, Yang J and Liu P: Loss of LKB1 disrupts breast epithelial cell polarity and promotes breast cancer metastasis and invasion. J Exp Clin Cancer Res. 33:702014. View Article : Google Scholar : PubMed/NCBI
28	Liang X, Wang P, Gao Q and Tao X: Exogenous activation of LKB1/AMPK signaling induces G(1) arrest in cells with endogenous LKB1 expression. Mol Med Rep. 9:1019–1024. 2014. View Article : Google Scholar : PubMed/NCBI
29	Luo L, Huang W, Tao R, Hu N, Xiao ZX and Luo Z: ATM and LKB1 dependent activation of AMPK sensitizes cancer cells to etoposide-induced apoptosis. Cancer Lett. 328:114–119. 2013. View Article : Google Scholar : PubMed/NCBI
30	Goodwin JM, Svensson RU, Lou HJ, Winslow MM, Turk BE and Shaw RJ: An AMPK-independent signaling pathway downstream of the LKB1 tumor suppressor controls Snail1 and metastatic potential. Mol Cell. 55:436–450. 2014. View Article : Google Scholar : PubMed/NCBI
31	Roy BC, Kohno T, Iwakawa R, Moriguchi T, Kiyono T, Morishita K, Sanchez-Cespedes M, Akiyama T and Yokota J: Involvement of LKB1 in epithelial-mesenchymal transition (EMT) of human lung cancer cells. Lung Cancer. 70:136–145. 2010. View Article : Google Scholar : PubMed/NCBI
32	Kopsiaftis S, Sullivan KL, Garg I, Taylor JR III and Claffey KP: AMPKα2 regulates bladder cancer growth through SKP2-mediated degradation of p27. Mol Cancer Res. 14:1182–1194. 2016. View Article : Google Scholar : PubMed/NCBI
33	Zhan P, Zhao S, Yan H, Yin C, Xiao Y, Wang Y, Ni R, Chen W, Wei G and Zhang P: α-enolase promotes tumorigenesis and metastasis via regulating AMPK/mTOR pathway in colorectal cancer. Mol Carcinog. 56:1427–1437. 2017. View Article : Google Scholar : PubMed/NCBI