Tetramethypyrazine inhibits renal cell carcinoma cells through inhibition of NKG2D signaling pathways

Luan,Yun; Liu,Junli; Liu,Xiaoli; Xue,Xia; Kong,Feng; Sun,Chao; Wang,Jue; Liu,Ling; Jia,Hongying

doi:10.3892/ijo.2016.3670

Tetramethypyrazine inhibits renal cell carcinoma cells through inhibition of NKG2D signaling pathways

Authors:
- Yun Luan
- Junli Liu
- Xiaoli Liu
- Xia Xue
- Feng Kong
- Chao Sun
- Jue Wang
- Ling Liu
- Hongying Jia
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Affiliations: Central Research Laboratory, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China, Clinical Molecular Biology Laboratory, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China, Department of Hematology, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China, Department of Pharmacy, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China, Center of Evidence-Based Medicine, The Second Hospital of Shandong University, Jinan, Shandong 250033, P.R. China
Published online on: August 23, 2016 https://doi.org/10.3892/ijo.2016.3670
Pages: 1704-1712

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Abstract

Tetramethypyrazine (TMP), one of the active compounds extracted from the traditional Chinese medicinal herb (Chuanxiong), has been verified as an anticancer compound against several types of cancer. However, understanding of the molecular mechanisms have not been fully elucidated. In the present study, the anticancer efficacy of TMP was investigated in human clear cell renal cell carcinoma (ccRCC) cells. We showed that TMP significantly inhibited ccRCC cell viability, proliferation, apoptosis, invasion and migration through the methods of MTT, flow cytometry, wound healing and transwell assays. Furthermore, reverse transcription polymerase chain reaction (RT-PCR), western blotting and immunofluorescence results demonstrated TMP upregulation of the expression of NKG2D ligands (NKG2DLs) MHC class I chain-related molecules A and B (MICA/B) and epithelial cell expression marker of E-cadherin, and downregulation of mesenchymal cell expression markers of vimentin and fibronectin. Taken together, the inhibition of TMP on ccRCC cells might be mediated via inhibition of NKG2D related signaling pathway to further suppress epithelial-mesenchymal transition (EMT) progression. The binding of NKG2D to its ligands activates NK cells, giving the rationale for studies on the utilization of TMP as a potential cancer therapeutic compound to increase NK cells-mediated cytotoxicity against high MICA/B expression in cancer cells.

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1	Yan Y, Zhao J, Cao C, Jia Z, Zhou N, Han S, Wang Y, Xu Y, Zhao J, Yan Y, et al: Tetramethylpyrazine promotes SH-SY5Y cell differentiation into neurons through epigenetic regulation of Topoisomerase IIβ. Neuroscience. 278:179–193. 2014. View Article : Google Scholar : PubMed/NCBI
2	Wang L, Zhang X, Cui G, Chan JY, Wang L, Li C, Shan L, Xu C, Zhang Q, Wang Y, et al: A novel agent exerts antitumor activity in breast cancer cells by targeting mitochondrial complex II. Oncotarget. Mar 27–2016.(Epub ahead of print).
3	Kao TK, Chang CY, Ou YC, Chen WY, Kuan YH, Pan HC, Liao SL, Li GZ and Chen CJ: Tetramethylpyrazine reduces cellular inflammatory response following permanent focal cerebral ischemia in rats. Exp Neurol. 247:188–201. 2013. View Article : Google Scholar : PubMed/NCBI
4	Liu X, Liu H, Zeng Z, Zhou W, Liu J and He Z: Pharmacokinetics of ligustrazine ethosome patch in rats and anti-myocardial ischemia and anti-ischemic reperfusion injury effect. Int J Nanomed. 6:1391–1398. 2011. View Article : Google Scholar
5	Jia Y, Wang Z, Zang A, Jiao S, Chen S and Fu Y: Tetramethylpyrazine inhibits tumor growth of lung cancer through disrupting angiogenesis via BMP/Smad/Id-1 signaling. Int J Oncol. 48:2079–2086. 2016.PubMed/NCBI
6	Kim M, Kim SO, Lee M, Lee JH, Jung WS, Moon SK, Kim YS, Cho KH, Ko CN and Lee EH: Tetramethylpyrazine, a natural alkaloid, attenuates pro-inflammatory mediators induced by amyloid β and interferon-γ in rat brain microglia. Eur J Pharmacol. 740:504–511. 2014. View Article : Google Scholar : PubMed/NCBI
7	Wang XJ, Xu YH, Yang GC, Chen HX and Zhang P: Tetramethylpyrazine inhibits the proliferation of acute lymphocytic leukemia cell lines via decrease in GSK-3β. Oncol Rep. 33:2368–2374. 2015.PubMed/NCBI
8	Zhang P, Zheng BB, Wang HY, Chen JH, Liu XY and Guo XL: DLJ14, a novel chemo-sensitization agent, enhances therapeutic effects of adriamycin against MCF-7/A cells both in vitro and in vivo. J Pharm Pharmacol. 66:398–407. 2014. View Article : Google Scholar : PubMed/NCBI
9	Yi B, Liu D, He M, Li Q, Liu T and Shao J: Role of the ROS/AMPK signaling pathway in tetramethylpyrazine-induced apoptosis in gastric cancer cells. Oncol Lett. 6:583–589. 2013.PubMed/NCBI
10	Chen Z, Pan X, Georgakilas AG, Chen P, Hu H, Yang Y, Tian S, Xia L, Zhang J, Cai X, et al: Tetramethylpyrazine (TMP) protects cerebral neurocytes and inhibits glioma by down regulating chemokine receptor CXCR4 expression. Cancer Lett. 336:281–289. 2013. View Article : Google Scholar : PubMed/NCBI
11	Zheng CY, Xiao W, Zhu MX, Pan XJ, Yang ZH and Zhou SY: Inhibition of cyclooxygenase-2 by tetramethylpyrazine and its effects on A549 cell invasion and metastasis. Int J Oncol. 40:2029–2037. 2012.PubMed/NCBI
12	Pan H, Jiang T, Cheng N, Wang Q, Ren S, Li X, Zhao C, Zhang L, Cai W and Zhou C: Long non-coding RNA BC087858 induces non-T790M mutation acquired resistance to EGFR-TKIs by activating PI3K/AKT and MEK/ERK pathways and EMT in non-small-cell lung cancer. Oncotarget. Jul 9–2016.(Epub ahead of print).
13	Bonyadi Rad E, Hammerlindl H, Wels C, Popper U, Ravindran Menon D, Breiteneder H, Kitzwoegerer M, Hafner C, Herlyn M, Bergler H, et al: Notch4 signaling induces a mesenchymal-epithelial-like transition in melanoma cells to suppress malignant behaviors. Cancer Res. 76:1690–1697. 2016. View Article : Google Scholar : PubMed/NCBI
14	Raza U, Saatci Ö, Uhlmann S, Ansari SA, Eyüpoğlu E, Yurdusev E, Mutlu M, Ersan PG, Altundağ MK and Zhang J: The miR-644a/CTBP1/p53 axis suppresses drug resistance by simultaneous inhibition of cell survival and epithelial-mesenchymal transition in breast cancer. Oncotarget. Jul 8–2016.(Epub ahead of print). PubMed/NCBI
15	Matusali G, Tchidjou HK, Pontrelli G, Bernardi S, D’ Ettorre G, Vullo V, Buonomini AR, Andreoni M, Santoni A, Cerboni C and Doria M: Soluble ligands for the NKG2D receptor are released during HIV-1 infection and impair NKG2D expression and cytotoxicity of NK cells. FASEB J. 27:2440–2450. 2013. View Article : Google Scholar : PubMed/NCBI
16	Guerra N, Tan YX, Joncker NT, Choy A, Gallardo F, Xiong N, Knoblaugh S, Cado D, Greenberg NM and Raulet DH: NKG2D-deficient mice are defective in tumor surveillance in models of spontaneous malignancy. Immunity. 28:571–580. 2008. View Article : Google Scholar : PubMed/NCBI
17	López-Larrea C, Suárez-Alvarez B, López-Soto A, López-Vázquez A and Gonzalez S: The NKG2D receptor: Sensing stressed cells. Trends Mol Med. 14:179–189. 2008. View Article : Google Scholar : PubMed/NCBI
18	Cosman D, Müllberg J, Sutherland CL, Chin W, Armitage R, Fanslow W, Kubin M and Chalupny NJ: ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. Immunity. 14:123–133. 2001. View Article : Google Scholar : PubMed/NCBI
19	López-Soto A, Zapico LH, Acebes-Huerta A, Rodrigo L and Gonzalez S: Regulation of NKG2D signaling during the epithelial-to-mesenchymal transition. Oncoimmunology. 2:e258202013. View Article : Google Scholar : PubMed/NCBI
20	Ni D, Ma X, Li HZ, Gao Y, Li XT, Zhang Y, Ai Q, Zhang P, Song EL, Huang QB, et al: Downregulation of FOXO3a promotes tumor metastasis and is associated with metastasis-free survival of patients with clear cell renal cell carcinoma. Clin Cancer Res. 20:1779–1790. 2014. View Article : Google Scholar : PubMed/NCBI
21	Jia HY, Liu JL, Yuan MZ, Zhou CJ, Sun WD, Zhao JJ, Wang J, Liu L and Luan Y: Regulation roles of MICA and NKG2D in human renal cancer cells. Asian Pac J Cancer Prev. 16:3901–3905. 2015. View Article : Google Scholar : PubMed/NCBI
22	Subramani R, Gonzalez E, Nandy SB, Arumugam A, Camacho F, Medel J, Alabi D and Lakshmanaswamy R: Gedunin inhibits pancreatic cancer by altering sonic hedgehog signaling pathway. Oncotarget. Mar 14–2016. View Article : Google Scholar : (Epub ahead of print). PubMed/NCBI
23	Friese MA, Wischhusen J, Wick W, Weiler M, Eisele G, Steinle A and Weller M: RNA interference targeting transforming growth factor-beta enhances NKG2D-mediated antiglioma immune response, inhibits glioma cell migration and invasiveness, and abrogates tumorigenicity in vivo. Cancer Res. 64:7596–7603. 2004. View Article : Google Scholar : PubMed/NCBI
24	Yang F, Shao Y, Yang F, Liu M, Huang J, Zhu K, Guo C, Luo J, Li W, Yang B, et al: Valproic acid upregulates NKG2D ligand expression and enhances susceptibility of human renal carcinoma cells to NK cell-mediated cytotoxicity. Arch Med Sci. 9:323–331. 2013. View Article : Google Scholar : PubMed/NCBI
25	Mikami S, Mizuno R, Kosaka T, Saya H, Oya M and Okada Y: Expression of TNF-α and CD44 is implicated in poor prognosis, cancer cell invasion, metastasis and resistance to the sunitinib treatment in clear cell renal cell carcinomas. Int J Cancer. 136:1504–1514. 2015. View Article : Google Scholar
26	Hill B, De Melo J, Yan J, Kapoor A, He L, Cutz JC, Feng X, Bakhtyar N and Tang D: Common reduction of the Raf kinase inhibitory protein in clear cell renal cell carcinoma. Oncotarget. 5:7406–7419. 2014. View Article : Google Scholar : PubMed/NCBI
27	Lawrie CH, Larrea E, Larrinaga G, Goicoechea I, Arestin M, Fernandez-Mercado M, Hes O, Cáceres F, Manterola L and López JI: Targeted next-generation sequencing and non-coding RNA expression analysis of clear cell papillary renal cell carcinoma suggests distinct pathological mechanisms from other renal tumour subtypes. J Pathol. 232:32–42. 2014. View Article : Google Scholar
28	Zhang X, Yan L, Jiao W, Ren J, Xing N, Zhang Y, Zang Y, Wang J and Xu Z: The clinical and biological significance of MICA in clear cell renal cell carcinoma patients. Tumour Biol. 37:2153–2159. 2016. View Article : Google Scholar
29	López-Soto A, Huergo-Zapico L, Galván JA, Rodrigo L, de Herreros AG, Astudillo A and Gonzalez S: Epithelial-mesenchymal transition induces an antitumor immune response mediated by NKG2D receptor. J Immunol. 190:4408–4419. 2013. View Article : Google Scholar : PubMed/NCBI
30	Eisele G, Wischhusen J, Mittelbronn M, Meyermann R, Waldhauer I, Steinle A, Weller M and Friese MA: TGF-β and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells. Brain. 129:2416–2425. 2006. View Article : Google Scholar : PubMed/NCBI
31	Kunigal S, Lakka SS, Joseph P, Estes N and Rao JS: Matrix metalloproteinase-9 inhibition down-regulates radiation-induced nuclear factor-kappa B activity leading to apoptosis in breast tumors. Clin Cancer Res. 14:3617–3626. 2008. View Article : Google Scholar : PubMed/NCBI