Expression of canonical transient receptor potential channels in U-2 OS and MNNG-HOS osteosarcoma cell lines

Lässig,Florian; Klann,Anja; Bekeschus,Sander; Lendeckel,Uwe; Wolke,Carmen

doi:10.3892/ol.2021.12568

Expression of canonical transient receptor potential channels in U-2 OS and MNNG-HOS osteosarcoma cell lines

Authors:
- Florian Lässig
- Anja Klann
- Sander Bekeschus
- Uwe Lendeckel
- Carmen Wolke
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Affiliations: Institute of Medical Biochemistry and Molecular Biology, University Medicine Greifswald, D-17475 Greifswald, Germany, Institute of Forensic Medicine, University Medicine Greifswald, D-17489, Greifswald, Germany, Zentrum für Innovationskompetenz (ZIK) plasmatis, Leibniz Institute for Plasma Science and Technology (INP), D-17489 Greifswald, Germany
Published online on: February 21, 2021 https://doi.org/10.3892/ol.2021.12568
Article Number: 307

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Abstract

In U-2 OS and MNNG-HOS osteosarcoma cells, small interfering RNA-mediated knockdown of the angiotensin-(1-7) receptor, Mas, increases cell proliferation. Whether alterations in canonical transient receptor potential channels (TRPC) expression contribute to this effect is not clear. In the present study, a basic description of TRPC subtype expression in osteosarcoma cell lines was provided. The pharmacological modulators of the angiotensin-(1-7) receptor, Mas, AVE0991 (agonist), or D-Ala⁷-Ang-(1-7) (antagonist) were applied to elucidate a possible role of Mas in the regulation of TRPC mRNA levels. The contribution of other G-protein coupled receptors (GPCR) or receptor tyrosine kinases to TRCP expression was studied by applying the selective pharmacological blockers of either PI3 kinase or MEK/Erk1/2 signaling, Ly294002 and PD98059. AVE0991 and D-Ala⁷-Ang-(1-7) exhibited no or marginal effects on TRPC mRNA expression. Ly294002 provoked a 9.6- and 5.9-fold increase in the amounts of TRPC5 mRNA in MNNG-HOS and U-2 OS cells, respectively. Additionally, Ly294002 increased TRPC6 mRNA levels; however, it had no effect on TRPCs 1, 3 and 4. Administration of PD98059 increased the amounts of TRPC6 and TRPC4 ~2-fold. In conclusion, the present study demonstrated that Mas-dependent alterations in osteosarcoma cell line proliferation were not mediated by any changes in TRPC subtype gene expression. The data shows in principle, and consistent with the literature, that the signaling pathways examined can regulate the expression of TRPCs at the mRNA level. Therefore, direct and signaling pathway-specific pharmacological targeting of TRPC subtypes may represent an option for improving the treatment of osteosarcoma.

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1	Gautier M, Dhennin-Duthille I, Ay AS, Rybarczyk P, Korichneva I and Ouadid-Ahidouch H: New insights into pharmacological tools to TR(i)P cancer up. Br J Pharmacol. 171:2582–2592. 2014. View Article : Google Scholar : PubMed/NCBI
2	Wu LJ, Sweet TB and Clapham DE: International Union of Basic and Clinical Pharmacology. LXXVI. Current progress in the mammalian TRP ion channel family. Pharmacol Rev. 62:381–404. 2010. View Article : Google Scholar : PubMed/NCBI
3	Capiod T: The need for calcium channels in cell proliferation. Recent Patents Anticancer Drug Discov. 8:4–17. 2013. View Article : Google Scholar
4	Hodeify R, Yu F, Courjaret R, Nader N, Dib M, Sun L, Adap E, Hubrack S and Machaca K: Regulation and role of store-operated Ca²⁺ entry in cellular proliferation. Calcium entry channels in non-excitable cells. Kozak JA and Putney JW Jr: CRC Press/Taylor & Francis; Boca Raton, FL: pp. 215–240. 2018
5	Bomben VC and Sontheimer HW: Inhibition of transient receptor potential canonical channels impairs cytokinesis in human malignant gliomas. Cell Prolif. 41:98–121. 2008. View Article : Google Scholar : PubMed/NCBI
6	Cai R, Ding X, Zhou K, Shi Y, Ge R, Ren G, Jin Y and Wang Y: Blockade of TRPC6 channels induced G2/M phase arrest and suppressed growth in human gastric cancer cells. Int J Cancer. 125:2281–2287. 2009. View Article : Google Scholar : PubMed/NCBI
7	Hwang JA, Hwang MK, Jang Y, Lee EJ, Kim JE, Oh MH, Shin DJ, Lim S, Ji G, Oh U, et al: 20-O-β-d-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginseng, inhibits colon cancer growth by targeting TRPC channel-mediated calcium influx. J Nutr Biochem. 24:1096–1104. 2013. View Article : Google Scholar : PubMed/NCBI
8	Jiang HN, Zeng B, Zhang Y, Daskoulidou N, Fan H, Qu JM and Xu SZ: Involvement of TRPC channels in lung cancer cell differentiation and the correlation analysis in human non-small cell lung cancer. PLoS One. 8:e676372013. View Article : Google Scholar : PubMed/NCBI
9	Gaunt HJ, Vasudev NS and Beech DJ: Transient receptor potential canonical 4 and 5 proteins as targets in cancer therapeutics. Eur Biophys J. 45:611–620. 2016. View Article : Google Scholar : PubMed/NCBI
10	Huang Z, Fan G and Wang D: Downregulation of calbindin 1, a calcium-binding protein, reduces the proliferation of osteosarcoma cells. Oncol Lett. 13:3727–3733. 2017. View Article : Google Scholar : PubMed/NCBI
11	Wang Y, Yang Z, Meng Z, Cao H, Zhu G, Liu T and Wang X: Knockdown of TRPM8 suppresses cancer malignancy and enhances epirubicin-induced apoptosis in human osteosarcoma cells. Int J Biol Sci. 10:90–102. 2013. View Article : Google Scholar : PubMed/NCBI
12	Berna-Erro A, Redondo PC and Rosado JA: Store-operated Ca²⁺ entry. Adv Exp Med Biol. 740:349–382. 2012. View Article : Google Scholar : PubMed/NCBI
13	Clapham DE, Runnels LW and Strübing C: The TRP ion channel family. Nat Rev Neurosci. 2:387–396. 2001. View Article : Google Scholar : PubMed/NCBI
14	Huang YW, Chang SJ, Harn HI, Huang HT, Lin HH, Shen MR, Tang MJ and Chiu WT: Mechanosensitive store-operated calcium entry regulates the formation of cell polarity. J Cell Physiol. 230:2086–2097. 2015. View Article : Google Scholar : PubMed/NCBI
15	Abed E, Labelle D, Martineau C, Loghin A and Moreau R: Expression of transient receptor potential (TRP) channels in human and murine osteoblast-like cells. Mol Membr Biol. 26:146–158. 2009. View Article : Google Scholar : PubMed/NCBI
16	Ender SA, Dallmer A, Lässig F, Lendeckel U and Wolke C: Expression and function of the ACE2/angiotensin(1–7)/Mas axis in osteosarcoma cell lines U-2 OS and MNNG-HOS. Mol Med Rep. 10:804–810. 2014. View Article : Google Scholar : PubMed/NCBI
17	Tirupula KC, Desnoyer R, Speth RC and Karnik SS: Atypical signaling and functional desensitization response of MAS receptor to peptide ligands. PLoS One. 9:e1035202014. View Article : Google Scholar : PubMed/NCBI
18	Sahr A, Wolke C, Maczewsky J, Krippeit-Drews P, Tetzner A, Drews G, Venz S, Gürtler S, van den Brandt J, Berg S, et al: The angiotensin-(1–7)/Mas axis improves pancreatic β-cell function in vitro and in vivo. Endocrinology. 157:4677–4690. 2016. View Article : Google Scholar : PubMed/NCBI
19	Chilukoti RK, Mostertz J, Bukowska A, Aderkast C, Felix SB, Busch M, Völker U, Goette A, Wolke C, Homuth G, et al: Effects of irbesartan on gene expression revealed by transcriptome analysis of left atrial tissue in a porcine model of acute rapid pacing in vivo. Int J Cardiol. 168:2100–2108. 2013. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−ΔΔC(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Chen J, Luan Y, Yu R, Zhang Z, Zhang J and Wang W: Transient receptor potential (TRP) channels, promising potential diagnostic and therapeutic tools for cancer. Biosci Trends. 8:1–10. 2014. View Article : Google Scholar : PubMed/NCBI
22	Bacsa B, Tiapko O, Stockner T and Groschner K: Mechanisms and significance of Ca²⁺ entry through TRPC channels. Curr Opin Physiol. 17:25–33. 2020. View Article : Google Scholar : PubMed/NCBI
23	Chen X, Sooch G, Demaree IS, White FA and Obukhov AG: Transient receptor potential canonical (TRPC) channels: Then and now. Cells. 9:2020. View Article : Google Scholar
24	Odell AF, Scott JL and Van Helden DF: Epidermal growth factor induces tyrosine phosphorylation, membrane insertion, and activation of transient receptor potential channel 4. J Biol Chem. 280:37974–37987. 2005. View Article : Google Scholar : PubMed/NCBI
25	Xu J, Wang H, Hu Y, Zhang YS, Wen L, Yin F, Wang Z, Zhang Y, Li S, Miao Y, et al: Inhibition of CaMKIIα activity enhances antitumor effect of fullerene C60 nanocrystals by suppression of autophagic degradation. Adv Sci (Weinh). 6:18012332019. View Article : Google Scholar : PubMed/NCBI
26	Asghar MY, Magnusson M, Kemppainen K, Sukumaran P, Löf C, Pulli I, Kalhori V and Törnquist K: Transient receptor potential canonical 1 (TRPC1) channels as Regulators of sphingolipid and VEGF receptor expression: Impolications for thyroid cancer cell migration and proliferation. J Biol Chem. 290:16116–16131. 2015. View Article : Google Scholar : PubMed/NCBI
27	Tao X, Zhao N, Jin H, Zhang Z, Liu Y, Wu J, Bast RC Jr, Yu Y and Feng Y: FSH enhances the proliferation of ovarian cancer cells by activating transient receptor potential channel C3. Endocr Relat Cancer. 20:415–429. 2013. View Article : Google Scholar : PubMed/NCBI
28	Shen B, Kwan HY, Ma X, Wong CO, Du J, Huang Y and Yao X: cAMP activates TRPC6 channels via the phosphatidylinositol 3-kinase (PI3K)-protein kinase B (PKB)-mitogen-activated protein kinase kinase (MEK)-ERK1/2 signaling pathway. J Biol Chem. 286:19439–19445. 2011. View Article : Google Scholar : PubMed/NCBI
29	Tetzner A, Gebolys K, Meinert C, Klein S, Uhlich A, Trebicka J, Villacañas Ó and Walther T: G-protein-coupled receptor MrgD is a receptor for angiotensin-(1–7) involving adenylyl cyclase, cAMP, and phosphokinase A. Hypertension. 68:185–194. 2016. View Article : Google Scholar : PubMed/NCBI
30	Thilo F, Liu Y, Loddenkemper C, Schuelein R, Schmidt A, Yan Z, Zhu Z, Zakrzewicz A, Gollasch M and Tepel M: VEGF regulates TRPC6 channels in podocytes. Nephrol Dial Transplant. 27:921–929. 2012. View Article : Google Scholar : PubMed/NCBI
31	Jardin I, Diez-Bello R, Lopez JJ, Redondo PC, Salido GM, Smani T and Rosado JA: TRPC6 channels are required for proliferation, migration and invasion of breast cancer cell lines by modulation of Orai1 and Orai3 surface exposure. Cancers (Basel). 10:3312018. View Article : Google Scholar
32	Heiser JH, Schuwald AM, Sillani G, Ye L, Müller WE and Leuner K: TRPC6 channel-mediated neurite outgrowth in PC12 cells and hippocampal neurons involves activation of RAS/MEK/ERK, PI3K, and CAMKIV signaling. J Neurochem. 127:303–313. 2013. View Article : Google Scholar : PubMed/NCBI
33	Numaga-Tomita T, Nishida M, Putney JW Jr and Mori Y: TRPC3 amplifies B-cell receptor-induced ERK signalling via protein kinase D-dependent Rap1 activation. Biochem J. 473:201–210. 2016. View Article : Google Scholar : PubMed/NCBI
34	Li X, Lu W, Fu X, Zhang Y, Yang K, Zhong N, Ran P and Wang J: BMP4 increases canonical transient receptor potential protein expression by activating p38 MAPK and ERK1/2 signaling pathways in pulmonary arterial smooth muscle cells. Am J Respir Cell Mol Biol. 49:212–220. 2013. View Article : Google Scholar : PubMed/NCBI