The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells

Özdil, Berrin; Aktug, Huseyin; Bozok Cetintas, Vildan; Doğan, Eda; Yildirim, Zafer; Duzgun, Zekeriya

The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells

Eda DOĞAN, (Ege Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, İzmir, Türkiye)

Zekeriya DÜZGÜN, (Giresun Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, Giresun, Türkiye)

Zafer YILDIRIM, (Ege Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji Anabilim Dalı, İzmir, Türkiye)

Berrin ÖZDİL, (Ege Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, İzmir, Türkiye)

Hüseyin AKTUĞ, (Ege Üniversitesi, Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, İzmir, Türkiye)

Vildan BOZOK ÇETİNTAŞ (Ege Üniversitesi, Tıp Fakültesi ,Tıbbi Biyoloji Anabilim Dalı, İzmir, Türkiye)

Turkish Journal of Biology

6 0

Yıl: 2021 Cilt: 45 Sayı: 1 Sayfa Aralığı: 26 - 34 Metin Dili: İngilizce İndeks Tarihi: 29-07-2022

The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells

Öz:

PIKfyve is an evolutionarily conserved lipid and protein kinase enzyme that has pleiotropic cellular functions. The aim of the present study was to investigate the effects of phosphatidylinositol-3-phosphate 5-kinase (PIKfyve) inhibitor, YM201636, on nonsmall cell lung cancer (NSCLC) cells growth, tumorigenicity, and claudin (CLDN) expressions. Three NSCLC cell lines (Calu-1, H1299 and HCC827) were used to compare the effects of YM201636. Cytotoxic effects of YM201636 were analysed using XTT assay. Malignancy potential of cells assesses with wound healing and soft agar colony-forming assays. mRNA and protein expressions of claudins were analysed by qRT-PCR and immunofluorescence staining. Our results revealed that YM201636 inhibited the proliferation and malignancy potential of Calu-1, H1299, and HCC827 cells in a dose-dependent manner. After YM201636 treatment CLDN1, -3 and -5 expressions increased significantly in HCC827 cells. CLDN3 and -5 expressions also significantly increased in Calu-1 cell line. YM201636 treatment significantly reduced the CLDN1 and increased the CLDN5 expression in H1299 cells. Immunofluorescence staining of CLDN1, -3 and -5 proteins showed a significant increase after YM201636 treatment. Besides, YM201636 induced EGFR mRNA expression in all NSCLC cell lines. Our results have shown that YM201636 inhibits tumorigenicity of NSCLC cells. Furthermore, estimated glomerular filtration rate (EGFR) pathway is important signalling involved in the regulation of claudins. Understanding the mechanisms of PIKfyve inhibitors may improve cancer treatment particularly for EGFR overactivated NSCLC.

Anahtar Kelime: Claudin epidermal growth factor receptor YM201636 PIKfyve nonsmall cell lung cancer

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

de Campos CB, Zhu YX, Sepetov N, Romanov S, Bruins LA et al. (2020). Identification of PIKfyve kinase as a target in multiple myeloma. Haematologica 105: 1641-1649.
Dukes JD, Whitley P, Chalmers AD (2012). The PIKfyve inhibitor YM201636 blocks the continuous recycling of the tight junction proteins claudin-1 and claudin-2 in MDCK cells. PLoS One 7:e28659.
Fang H, Wang Y, Xu L, Zhou S, Bai J et al. (2019). EGFR inhibitor gefitinib regulates barrier function in human epidermal keratinocytes via the modulation of the expression of claudins. International Journal of Molecular Medicine 43: 1522-1530.
Hou JZ, Xi ZQ, Niu J, Li W, Wang X, et al. (2019). Inhibition of PIKfyve using YM201636 suppresses the growth of liver cancer via the induction of autophagy. Oncology Reports 41:1971- 1979.
Hung CY, Yang WB, Wang SA, Hsu TI, Chang WC, et al. (2014). Nucleolin enhances internal ribosomal entry site (IRES)- mediated translation of Sp1 in tumorigenesis. Biochimica et Biophysica Acta 1843:2843-2854.
Ikonomov OC, Altankov G, Sbrissa D, Shisheva A (2018). PIKfyve inhibitor cytotoxicity requires AKT suppression and excessive cytoplasmic vacuolation. Toxicology and Applied Pharmacology 356: 151-158.
Ikonomov OC, Sbrissa D, Shisheva A (2006). Localized PtdIns 3,5- P2 synthesis to regulate early endosome dynamics and fusion. American Journal of Physiology-Cell Physiology 291: C393- 404. J efferies HB, Cooke FT, Jat P, Boucheron C, Koizumi T et al. (2008). A selective PIKfyve inhibitor blocks PtdIns(3,5)P(2) production and disrupts endomembrane transport and retroviral budding. EMBO Reports 9: 164-170.
Khan N, Asif AR (2015). Transcriptional regulators of claudins in epithelial tight junctions. Mediators of Inflammation 2015: 219843.
Kim H, Kim SH, Hwang D, An J, Chung HS, et al. (2020). Extracellular pyruvate kinase M2 facilitates cell migration by upregulating claudin-1 expression in colon cancer cells. Biochemistry and Cell Biology 98: 219-226.
Kim J, Jahng WJ, Di Vizio D, Lee JS, Jhaveri R et al . (2007). The phosphoinositide kinase PIKfyve mediates epidermal growth factor receptor trafficking to the nucleus. Cancer Research 67: 9229-9237.
Krause G, Winkler L, Piehl C, Blasig I, Piontek J et al . (2009). Structure and function of extracellular claudin domains. Annals of the New York Academy of Sciences 1165: 34-43.
Ma L, Yin W, Ma H, Elshoura I, Wang L (2019). Targeting claudin-3 suppresses stem cell-like phenotype in nonsquamous nonsmall-cell lung carcinoma. Lung Cancer Management 8:LMT04.
Martin S, Harper CB, May LM, Coulson EJ, Meunier FA et al . (2013). Inhibition of PIKfyve by YM-201636 dysregulates autophagy and leads to apoptosis-independent neuronal cell death. PLoS One 8:e60152.
Martin TA, Jiang WG (2009). Loss of tight junction barrier function and its role in cancer metastasis. Biochimica et Biophysica Acta 1788: 872-891.
McCartney AJ, Zolov SN, Kauffman EJ, Zhang Y, Strunk BS et al . (2014). Activity-dependent PI(3,5)P2 synthesis controls AMPA receptor trafficking during synaptic depression. Proceedings of the National Academy of Sciences 111:E4896-4905.
Merikallio H, Kaarteenaho R, Paakko P, Lehtonen S, Hirvikoski P et al . (2011). Impact of smoking on the expression of claudins in lung carcinoma. European Journal of Cancer 47: 620-630.
Moldvay J, Fabian K, Jackel M, Nemeth Z, Bogos K et al . (2017). Claudin-1 Protein Expression Is a Good Prognostic Factor in Non-Small Cell Lung Cancer, but only in Squamous Cell Carcinoma Cases. Pathology & Oncology Research 23: 151- 156.
Moldvay J, Jackel M, Paska C, Soltesz I, Schaff Z et al . (2007). Distinct claudin expression profile in histologic subtypes of lung cancer. Lung Cancer 57: 159-167.
Oppelt A, Haugsten EM, Zech T, Danielsen HE, Sveen A et al . (2014). PIKfyve, MTMR3 and their product PtdIns5P regulate cancer cell migration and invasion through activation of Rac1. Biochemical Journal 461: 383-390.
Oppelt A, Lobert VH, Haglund K, Mackey AM, Rameh LE et al . (2013). Production of phosphatidylinositol 5-phosphate via PIKfyve and MTMR3 regulates cell migration. EMBO Reports 14: 57-64.
Salvador E, Burek M, Forster CY (2016). Tight Junctions and the Tumor Microenvironment. Current Pathobiology Reports 4: 135-145.
Sbrissa D, Ikonomov OC, Fu Z, Ijuin T, Gruenberg J et al . (2007). Core protein machinery for mammalian phosphatidylinositol 3,5-bisphosphate synthesis and turnover that regulates the progression of endosomal transport. Novel Sac phosphatase joins the ArPIKfyve-PIKfyve complex. Journal of Biological Chemistry 282: 23878-23891.
Sbrissa D, Ikonomov OC, Strakova J, Shisheva A (2004). Role for a novel signaling intermediate, phosphatidylinositol 5-phosphate, in insulin-regulated F-actin stress fiber breakdown and GLUT4 translocation. Endocrinology 145: 4853-4865.
Shin K, Fogg VC, Margolis B (2006). Tight junctions and cell polarity. Annual Review of Cell and Developmental Biology 22: 207-235.
Shisheva A (2001). PIKfyve: the road to PtdIns 5-P and PtdIns 3,5- P(2). Cell Biology International 25:1201-1206.
Shisheva A (2008). PIKfyve: Partners, significance, debates and paradoxes. Cell Biology International 32: 591-604.
Siegel RL, Miller KD, Jemal A (2020). Cancer statistics, 2020. CA: A Cancer Journal for Clinicians 70: 7-30.
Singh AB, Harris RC (2004). Epidermal growth factor receptor activation differentially regulates claudin expression and enhances transepithelial resistance in Madin-Darby canine kidney cells. Journal of Biological Chemistry 279: 3543-3552.
Soini Y (2011). Claudins in lung diseases. Respiratory Research 12: 70.
Sun BS, Yao YQ, Pei BX, Zhang ZF, Wang CL (2016). Claudin-1 correlates with poor prognosis in lung adenocarcinoma. Thoracic Cancer 7: 556-563.
Tan X, Thapa N, Sun Y, Anderson RA (2015). A kinase-independent role for EGF receptor in autophagy initiation. Cell 160: 145- 160.
Zhang L, Wang Y, Zhang B, Zhang H, Zhou M et al . (2017). Claudin-3 expression increases the malignant potential of lung adenocarcinoma cells: role of epidermal growth factor receptor activation. Oncotarget 8: 23033-23047.

APA	Doğan E, Duzgun Z, Yildirim Z, Özdil B, Aktug H, Bozok Cetintas V (2021). The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. , 26 - 34.
Chicago	Doğan Eda,Duzgun Zekeriya,Yildirim Zafer,Özdil Berrin,Aktug Huseyin,Bozok Cetintas Vildan The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. (2021): 26 - 34.
MLA	Doğan Eda,Duzgun Zekeriya,Yildirim Zafer,Özdil Berrin,Aktug Huseyin,Bozok Cetintas Vildan The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. , 2021, ss.26 - 34.
AMA	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. . 2021; 26 - 34.
Vancouver	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. . 2021; 26 - 34.
IEEE	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V "The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells." , ss.26 - 34, 2021.
ISNAD	Doğan, Eda vd. "The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells". (2021), 26-34.

APA	Doğan E, Duzgun Z, Yildirim Z, Özdil B, Aktug H, Bozok Cetintas V (2021). The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. Turkish Journal of Biology, 45(1), 26 - 34.
Chicago	Doğan Eda,Duzgun Zekeriya,Yildirim Zafer,Özdil Berrin,Aktug Huseyin,Bozok Cetintas Vildan The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. Turkish Journal of Biology 45, no.1 (2021): 26 - 34.
MLA	Doğan Eda,Duzgun Zekeriya,Yildirim Zafer,Özdil Berrin,Aktug Huseyin,Bozok Cetintas Vildan The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. Turkish Journal of Biology, vol.45, no.1, 2021, ss.26 - 34.
AMA	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. Turkish Journal of Biology. 2021; 45(1): 26 - 34.
Vancouver	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells. Turkish Journal of Biology. 2021; 45(1): 26 - 34.
IEEE	Doğan E,Duzgun Z,Yildirim Z,Özdil B,Aktug H,Bozok Cetintas V "The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells." Turkish Journal of Biology, 45, ss.26 - 34, 2021.
ISNAD	Doğan, Eda vd. "The effects of PIKfyve inhibitor YM201636 on claudins and malignancy potential of nonsmall cell cancer cells". Turkish Journal of Biology 45/1 (2021), 26-34.