Tunicamycin induced inhibition of calpain 1 and 2 enzyme activity in ovarian cancer cells

Seda M. Sari Kilicaslan; Zerrin İncesu

doi:10.18203/2319-2003.ijbcp20230689

Authors

Seda M. Sari Kilicaslan Faculty of Education, Anadolu University, Eskisehir, Turkey
Zerrin İncesu Department of Biochemistry Science, Faculty of Pharmacy, Anadolu University, Eskisehir, Turkey

DOI:

https://doi.org/10.18203/2319-2003.ijbcp20230689

Keywords:

IHOSE, SKOV-3, Fibronectin, Calpain

Abstract

Background: Tunicamycin (TN) is an antitumor agent and induced intracellular calcium levels in many cells, however its molecular mechanism is still needed to be explored. Calpeptin (Calp) is an inhibitor of both calpain 1 and 2 (CAPN-1/2) enzymes, and plays a fundamental role in tumor mechanism. In this study, the effects of TN and Calp were investigated on CAPN-1/2 enzyme activity in normal and ovarian cancer cells adhered to fibronectin.

Methods: 24uM TN, 50uM Calp, and combined TN and Calp (TN+Calp) were applied for 1 and 12 hours to FN-bound (FN+) and non-FN-bound (FN-) normal human ovarian epithelial (IHOSE) and ovarian cancer (SKOV-3) cells. The activation of CAPN-1/2 was measured by the luminescent method and the significance of the results was analyzed with the t-test.

Results: CAPN-1/2 enzyme activity (at 12 hour) was present in both cell lines, but the level of enzyme is higher in IHOSE cells compared to SKOV-3 cells. The results showed that 1 hour TN and TN+Calp applications stimulated CAPN-1/2 enzyme activity in IHOSE cells but did not show any stimulating effect in SKOV-3 cells. After 12-hour of treatment, the cells with TN, Calp or TN+Calp showed an inhibitory effect on CAPN-1/2 enzyme in both FN+IHOSE and SKOV-3 cells. At 12-hour TN+Calp administration was determined to be the most effective inhibitor in FN+ SKOV-3 since it inhibited CAPN1/2 activity statistically significantly more than both Calp and TN administrations.

Conclusions: The effects of TN, Calp and TN+Calp applications on the CAPN-1/2 enzyme varied according to the cell type, normal or cancer cells, and whether the cell was bound to FN and the incubation period. 12 h administrations of TN, Calp or TN+Calp inhibited the CAPN-1/2 enzyme in both FN+ IHOSE and SKOV-3 cells.

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References

Kovacs L, Su Y. The Critical Role of Calpain in Cell Proliferation. J Biomol Res Ther. 2014;3(3):1000112.

Carragher ON, Frame CM. Calpain: a role in cell transformation and migration. Int J of Biochem Cell Biol. 2002;34:1539-43.

Chen J, Wu Y, Zhang L, Fang X, Hu X. Evidence for calpains in cancer metastasis. J Cell Physiol. 2019;234(6):8233-40.

Kondakova IV, Yunusova NV, Spirina LV, Kolomiets LA, Villert AB. Association between intracellular proteinase activities and the content of locomotor proteins in tissues of primary tumors and metastases of ovarian cancer. Russ J Bioorg Chem. 2014;40:681-7.

Storr SJ, Safuan S, Woolston CM, Abdel-Fatah T, Deen S, Chan SY, Martin SG. Calpain-2 expression is associated with response to platinum based chemotherapy, progression-free and overall survival in ovarian cancer. J Cellular Mol Med. 2012;16:2422-8.

Salehin D, Fromberg I, Haugk C, Dohmen B, Georg T, Bohle RM, et al. Immunhistochemical analysis for expression of calpain 1, calpain 2 and calpastatin in ovarian cancer. Eur J Gynaecol Oncol. 2011;32(6):628-35.

Zhang S, Deen S, Storr SJ, Chondrou PS, Nicholls H, Yao A, et al. Calpain system protein expression and activity in ovarian cancer. J Cancer Res Clin Oncol. 2019;145(2):345-61.

Shapovalov I, Harper D, Greer PA. Calpain as a therapeutic target in cancer. Expert Opin Ther Targets. 2022;26(3):217-31.

Buckley BJ, Whorton AR. Tunicamycin increases intracellular calcium levels in bovine aortic endothelial cells. Am J Physiol. 1997;273(4):C1298-305.

Guha P, Kaptan E, Gade P, Kalvakolanu DV, Ahmed H. Tunicamycin induced endoplasmic reticulum stress promotes apoptosis of prostate cancer cells by activating mTORC1. Oncotarget. 2017;15;8(40):68191-207.

Kratochvílová K, Horak P, Ešner M, Souček K, Pils D, Anees M, et al. Tumor suppressor candidate 3 (TUSC3) prevents the epithelial-to-mesenchymal transition and inhibits tumor growth by modulating the endoplasmic reticulum stress response in ovarian cancer cells. Int J Cancer. 2015;15;137(6):1330-40.

Fang J, Liu X, Bolanos L, Barker B, Rigolino C, Cortelezzi A, et al. A calcium- and calpain-dependent pathway determines the response to lenalidomide in myelodysplastic syndromes. Nat Med. 2016;22(7):727-34.

Ono Y, Saido TC, Sorimachi H. Calpain research for drug discovery: challenges and potential. Nat Rev Drug Discov. 2016;15(12):854-76.

Chen B, Tang J, Guo YS, Li Y, Chen ZN, Jiang JL. Calpains are required for invasive and metastatic potentials of human HCC cells. Cell Biol Int. 2013;37(7):643-52.

Li CL, Yang D, Cao X, Wang F, Hong DY, Wang J, Shen XC, Chen Y. Fibronectin induces epithelial-mesenchymal transition in human breast cancer MCF-7 cells via activation of calpain. Oncol Lett. 2017;13(5):3889-95.

Chen Q, Thompson J, Hu Y, Lesnefsky EJ. Tunicamycin-Induced Endoplasmic Reticulum Stress Damages Complex I in Cardiac Mitochondria. Life (Basel). 2022;12(8):1209.

Hou H, Ge C, Sun H, Li H, Li J, Tian H. Tunicamycin inhibits cell proliferation and migration in hepatocellular carcinoma through suppression of CD44s and the ERK1/2 pathway. Cancer Sci. 2018;109(4):1088-100.

Rose AH, Huang Z, Mafnas C, Hara JH, Hoffmann FW, Hashimoto AS, et al. Calpain-2 Inhibitor Therapy Reduces Murine Colitis and Colitis- associated Cancer Inflamm Bowel Dis. 2015;21(9):2005-15.

Lu X, Li G, Liu S, Wang H, Chen B. MiR-585-3p suppresses tumor proliferation and Migration by directly targeting CAPN9 in high grade serous ovarian cancer. J Ovarian Res. 2021;14,90.