Identification of the role of TRPM8 in glioblastoma and its effect on proliferation, apoptosis and invasion of the U251 human glioblastoma cell line

  • Authors:
    • Jianping Zeng
    • Ye Wu
    • Siyi Zhuang
    • Liping Qin
    • Shushan Hua
    • Rajneesh Mungur
    • Jianwei Pan
    • Yu Zhu
    • Renya Zhan
  • View Affiliations

  • Published online on: August 5, 2019     https://doi.org/10.3892/or.2019.7260
  • Pages: 1517-1526
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Abstract

Glioblastoma multiforme (GBM) is the most commonly occurring brain cancer, and is characterized by its poor patient outcomes. The present study examined the mRNA expression levels of the transient receptor potential melastatin (TRPM) family in various types of cancer using the ONCOMINE database, along with their corresponding expression profiles in an array of cancer cell lines based on the Cancer Cell Line Encyclopedia (CCLE) datasets. Kaplan‑Meier plotter survival analysis via the Chinese Glioma Genome Atlas (CGGA) database was also used to evaluate the prognostic value of transient receptor potential melastatin 8 (TRPM8). For the activity test on the TRPM8 channel, patch‑clamp recordings and Ca2+ measurements by fluorescence imaging of Fluo‑4am were performed. Short hairpin RNA (shRNA) targeting TRPM8 was designed, synthesized and then transfected into the U251 cells via Lipofectamine 2000. The expression of extracellular singnal‑regulated kinase (ERK), cyclin D1 and Bcl‑2 were detected by performing western blotting and immunofluorescence. The apoptosis, proliferation and invasion of glioma cells were detected by using flow cytometry, and CCK‑8 and Transwell invasion assays. In the present study, TRPM8 was distinctively upregulated in GBM cell lines. TRPM8 is functional and has the characteristic of outward rectification, which was verified via electrophysiology and Ca2+ fluorescence imaging in U251 cells. The western blot and immunofluorescence results revealed that the expression of ERK, cyclin D1 and Bcl‑2 were decreased in the shRNA interference group. The CCK‑8 assay demonstrated that the proliferation ability of U251 cells in the U251/TRPM8 group was higher than that in the U251 group and U251/Con group (P<0.05). The result of the Transwell invasion assay indicated that the invasion of human glioblastoma U251 cells was positively correlated with the expression level of TRPM8. Collectively, the results of the present study indicated that Ca2+‑permeable TRPM8 nonselective cation channels contribute to survival, proliferation, apoptosis, and local tumor invasion of glioblastoma. Therefore, TRPM8 is a promising biomarker for aggressiveness of GBM, and a potential target in future anti‑glioblastoma therapies.

References

1 

Louis DN: Molecular pathology of malignant gliomas. Annu Rev Pathol. 1:97–117. 2006. View Article : Google Scholar : PubMed/NCBI

2 

Jiang T, Mao Y, Ma W, Mao Q, You Y, Yang X, Jiang C, Kang C, Li X, Chen L, et al: CGCG clinical practice guidelines for the management of adult diffuse gliomas. Cancer Lett. 375:263–273. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Li MY, Yang P, Liu YW, Zhang CB, Wang KY, Wang YY, Yao K, Zhang W, Qiu XG, Li WB, et al: Low c-Met expression levels are prognostic for and predict the benefits of temozolomide chemotherapy in malignant gliomas. Sci Rep. 6:211412016. View Article : Google Scholar : PubMed/NCBI

4 

Stupp R, Dietrich PY, Ostermann Kraljevic S, Pica A, Maillard I, Maeder P, Meuli R, Janzer R, Pizzolato G, Miralbell R, et al: Promising survival for patients with newly diagnosed glioblastoma multiforme treated with concomitant radiation plus temozolomide followed by adjuvant temozolomide. J Clin Oncol. 20:1375–1382. 2002. View Article : Google Scholar : PubMed/NCBI

5 

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

6 

Wetsel WC: Sensing hot and cold with TRP channels. Int J Hyperthermia. 27:388–398. 2011. View Article : Google Scholar : PubMed/NCBI

7 

Park YR, Chun JN, So I, Kim HJ, Baek S, Jeon JH and Shin SY: Data-driven analysis of TRP channels in cancer: Linking variation in gene expression to clinical significance. Cancer Genomics Proteomics. 13:83–90. 2016.PubMed/NCBI

8 

Liu Z, Wu H, Wei Z, Wang X, Shen P, Wang S, Wang A, Chen W and Lu Y: TRPM8: A potential target for cancer treatment. J Cancer Res Clin Oncol. 142:1871–1881. 2016. View Article : Google Scholar : PubMed/NCBI

9 

Alptekin M, Eroglu S, Tutar E, Sencan S, Geyik MA, Ulasli M, Demiryurek AT and Camci C: Gene expressions of TRP channels in glioblastoma multiforme and relation with survival. Tumour Biol. 36:9209–9213. 2015. View Article : Google Scholar : PubMed/NCBI

10 

Zhang L and Barritt GJ: TRPM8 in prostate cancer cells: A potential diagnostic and prognostic marker with a secretory function? Endocr Relat Cancer. 13:27–38. 2006. View Article : Google Scholar : PubMed/NCBI

11 

Burke RC, Bardet SM, Carr L, Romanenko S, Arnaud-Cormos D, Leveque P and O'Connor RP: Nanosecond pulsed electric fields depolarize transmembrane potential via voltage-gated K+, Ca2+ and TRPM8 channels in U87 glioblastoma cells. Biochim Biophys Acta Biomembr. 1859:2040–2050. 2017. View Article : Google Scholar : PubMed/NCBI

12 

Yang M, Li Y, Chilukuri K, Brady OA, Boulos MI, Kappes JC and Galileo DS: L1 stimulation of human glioma cell motility correlates with FAK activation. J Neurooncol. 105:27–44. 2011. View Article : Google Scholar : PubMed/NCBI

13 

Polivka J Jr, Polivka J, Holubec L, Kubikova T, Priban V, Hes O, Pivovarcikova K and Treskova I: Advances in experimental targeted therapy and immunotherapy for patients with glioblastoma multiforme. Anticancer Res. 37:21–33. 2017. View Article : Google Scholar : PubMed/NCBI

14 

Stoyanov GS and Dzhenkov DL: On the concepts and history of glioblastoma Multiforme-Morphology, genetics and epigenetics. Folia Med (Plovdiv). 60:48–66. 2018. View Article : Google Scholar : PubMed/NCBI

15 

Jhaveri N, Chen TC and Hofman FM: Tumor vasculature and glioma stem cells: Contributions to glioma progression. Cancer Lett. 380:545–551. 2016. View Article : Google Scholar : PubMed/NCBI

16 

Hardee ME and Zagzag D: Mechanisms of glioma-associated neovascularization. Am J Pathol. 181:1126–1141. 2012. View Article : Google Scholar : PubMed/NCBI

17 

Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJ, Belanger K, Brandes AA, Marosi C, Bogdahn U, et al: Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. New Engl J Med. 352:987–996. 2005. View Article : Google Scholar : PubMed/NCBI

18 

Sturm D, Bender S, Jones DT, Lichter P, Grill J, Becher O, Hawkins C, Majewski J, Jones C, Costello JF, et al: Paediatric and adult glioblastoma: Multiform (epi)genomic culprits emerge. Nat Rev Cancer. 14:92–107. 2014. View Article : Google Scholar : PubMed/NCBI

19 

Roderick HL and Cook SJ: Ca2+ signalling checkpoints in cancer: Remodelling Ca2+ for cancer cell proliferation and survival. Nat Rev Cancer. 8:361–375. 2008. View Article : Google Scholar : PubMed/NCBI

20 

Sun J, Mu H, Dai K and Yi L: Calreticulin: A potential anti-cancer therapeutic target. Pharmazie. 72:503–510. 2017.PubMed/NCBI

21 

Ohshima Y, Takata N, Suzuki-Karasaki M, Yoshida Y, Tokuhashi Y and Suzuki-Karasaki Y: Disrupting mitochondrial Ca2+ homeostasis causes tumor-selective TRAIL sensitization through mitochondrial network abnormalities. Int J Oncol. 51:1146–1158. 2017. View Article : Google Scholar : PubMed/NCBI

22 

Rizzuto R, Pinton P, Ferrari D, Chami M, Szabadkai G, Magalhães PJ, Di Virgilio F and Pozzan T: Calcium and apoptosis: Facts and hypotheses. Oncogene. 22:8619–8627. 2003. View Article : Google Scholar : PubMed/NCBI

23 

Fiorio Pla A and Gkika D: Emerging role of TRP channels in cell migration: From tumor vascularization to metastasis. Front Physiol. 4:3112013. View Article : Google Scholar : PubMed/NCBI

24 

Hecquet CM, Zhang M, Mittal M, Vogel SM, Di A, Gao X, Bonini MG and Malik AB: Cooperative interaction of trp Melastatin channel transient receptor potential (TRPM2) with its splice variant TRPM2 Short variant is essential for endothelial cell apoptosis. Circ Res. 114:469–479. 2014. View Article : Google Scholar : PubMed/NCBI

25 

Landsberg JW and Yuan JX: Calcium and TRP channels in pulmonary vascular smooth muscle cell proliferation. News Physiol Sci. 19:44–50. 2004.PubMed/NCBI

26 

Hantute-Ghesquier A, Haustrate A, Prevarskaya N and Lehen'kyi V: TRPM family channels in cancer. Pharmaceuticals (Basel). 11(pii): E582018. View Article : Google Scholar : PubMed/NCBI

27 

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

28 

Wong KK, Banham AH, Yaacob NS and Nur Husna SM: The oncogenic roles of TRPM ion channels in cancer. J Cell Physiol. Feb 2–2019.doi: 10.1002/jcp.28168 (Epub ahead of print).

29 

Okamoto Y, Ohkubo T, Ikebe T and Yamazaki J: Blockade of TRPM8 activity reduces the invasion potential of oral squamous carcinoma cell lines. Int J Oncol. 40:1431–1440. 2012.PubMed/NCBI

30 

Kijpornyongpan T, Sereemaspun A and Chanchao C: Dose-dependent cytotoxic effects of menthol on human malignant melanoma A-375 cells: Correlation with TRPM8 transcript expression. Asian Pac J Cancer Prev. 15:1551–1556. 2014. View Article : Google Scholar : PubMed/NCBI

31 

Gkika D and Prevarskaya N: Molecular mechanisms of TRP regulation in tumor growth and metastasis. Biochim Biophys Acta. 1793:953–958. 2009. View Article : Google Scholar : PubMed/NCBI

32 

Sideris M, Emin EI, Abdullah Z, Hanrahan J, Stefatou KM, Sevas V, Emin E, Hollingworth T, Odejinmi F, Papagrigoriadis S, et al: The role of KRAS in endometrial cancer: A mini-review. Anticancer Res. 39:533–539. 2019. View Article : Google Scholar : PubMed/NCBI

33 

Liu F, Yang X, Geng M and Huang M: Targeting ERK, an Achilles' Heel of the MAPK pathway, in cancer therapy. Acta Pharm Sin B. 8:552–562. 2018. View Article : Google Scholar : PubMed/NCBI

34 

Sugden PH and Clerk A: Regulation of the ERK subgroup of MAP kinase cascades through G protein-coupled receptors. Cell Signal. 9:337–351. 1997. View Article : Google Scholar : PubMed/NCBI

35 

Krueger JS, Keshamouni VG, Atanaskova N and Reddy KB: Temporal and quantitative regulation of mitogen-activated protein kinase (MAPK) modulates cell motility and invasion. Oncogene. 20:4209–4218. 2001. View Article : Google Scholar : PubMed/NCBI

36 

Samatar AA and Poulikakos PI: Targeting RAS-ERK signalling in cancer: Promises and challenges. Nat Rev Drug Discov. 13:928–942. 2014. View Article : Google Scholar : PubMed/NCBI

37 

Cobb MH, Hepler JE, Cheng M and Robbins D: The mitogen-activated protein kinases, ERK1 and ERK2. Semin Cancer Biol. 5:261–268. 1994.PubMed/NCBI

38 

Chen WT, Hsu FT, Liu YC, Chen CH, Hsu LC and Lin SS: Fluoxetine induces apoptosis through extrinsic/intrinsic pathways and inhibits ERK/NF-κB-modulated anti-apoptotic and invasive potential in hepatocellular carcinoma cells in vitro. Int J Mol Sci. 20(pii): E7572019. View Article : Google Scholar : PubMed/NCBI

39 

McIlwain DR, Berger T and Mak TW: Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol. 5:a0086562013. View Article : Google Scholar : PubMed/NCBI

40 

Elmore S: Apoptosis: A review of programmed cell death. Toxicol Pathol. 35:495–516. 2007. View Article : Google Scholar : PubMed/NCBI

41 

Wondergem R, Ecay TW, Mahieu F, Owsianik G and Nilius B: HGF/SF and menthol increase human glioblastoma cell calcium and migration. Biochem Biophys Res Commun. 372:210–215. 2008. View Article : Google Scholar : PubMed/NCBI

42 

Wondergem R and Bartley JW: Menthol increases human glioblastoma intracellular Ca2+, BK channel activity and cell migration. J Biomed Sci. 16:902009. View Article : Google Scholar : PubMed/NCBI

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October 2019
Volume 42 Issue 4

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Copy and paste a formatted citation
APA
Zeng, J., Wu, Y., Zhuang, S., Qin, L., Hua, S., Mungur, R. ... Zhan, R. (2019). Identification of the role of TRPM8 in glioblastoma and its effect on proliferation, apoptosis and invasion of the U251 human glioblastoma cell line. Oncology Reports, 42, 1517-1526. https://doi.org/10.3892/or.2019.7260
MLA
Zeng, J., Wu, Y., Zhuang, S., Qin, L., Hua, S., Mungur, R., Pan, J., Zhu, Y., Zhan, R."Identification of the role of TRPM8 in glioblastoma and its effect on proliferation, apoptosis and invasion of the U251 human glioblastoma cell line". Oncology Reports 42.4 (2019): 1517-1526.
Chicago
Zeng, J., Wu, Y., Zhuang, S., Qin, L., Hua, S., Mungur, R., Pan, J., Zhu, Y., Zhan, R."Identification of the role of TRPM8 in glioblastoma and its effect on proliferation, apoptosis and invasion of the U251 human glioblastoma cell line". Oncology Reports 42, no. 4 (2019): 1517-1526. https://doi.org/10.3892/or.2019.7260