The effects of Tilia (Tilia plathyphyllos) extract on gene expression of caspase 3 and caspase 9 in canine mammary gland cancer cell line (CF41.Mg)

Subject Areas : Veterinary Clinical Pathology

Zohreh Mostahsan ¹ (-D.V.M. Graduate, Faculty of Specialized Veterinary Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.)
Pejhman Mortazavi ² (Associate Professor, Department of Pathology, Faculty of Specialized Veterinary Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.)

Received: 2017-04-20 Accepted : 2017-10-04 Published : 2019-04-21

Keywords: Caspase 3, Tilia extract, Caspase 9, Canine mammary gland cancer cell line,

Abstract :

Mammary gland tumors arethe most common type of cancer in dogs and cats after skin tumors. Any disorder in the process of apoptosis causes the growth of cancer cells. Caspases 3 and 9 play an important role in the process of apoptosis as initiator and executioner caspases. This study was conducted to investigate the effect of Tilia (Tilia plathyphyllos) extract on gene expression of caspase 3 and caspase 9 in canine mammary gland cancer cell line. Canine mammary gland tumor cells (CF41.Mg) were passaged in the presence of 5, 10, 25, 50 and 100 mg/ml of Tilia extract for 24, 48 and 72 hours. Microculture tetrajolium test (MTT) was used after the passage to assay the number of live cells. To understand the mechanisms involved in induction of cell death at previously mentioned intervals, RNA was extracted and cDNA synthetizedand the rate of gene expression of caspase 3 and 9 was evaluated using specific primers with RT-PCR. According to the result, maximum  anti-tumor activity of this extract was observed after 48 hours with the  dose of 25 mg/ml. RT-PCR results indicated gene expression of caspase 3 and 9 after 24, 48 and 72 hours in the treated samples. The highest extent of gene expression was observed at 10 and 100 mg/ml of extract for caspase 3 and 10 mg/ml of extract for caspase 9. The results of the present study confirmed the inhibitory effects of Tilia extract on growth of canine mammary gland tumor cell line.

References:

• Brizi, M.R., Marrassini, C., Zettler, G., Ferraro, G., Anesini, C. (2012). Comparative antiproliferative action of two extracts from Tilia x viridis on normal and tumoral lymphocytes: relationship with antioxidant activity. Chinese Medicine, 3: 20-29.


• Cassali, G.D., Lavalle, G.E., De Nardi, A.B., Ferreira, E., Bertagnolli, A.C., Estrela-Lima, A., et al. (2011). Consensus for the Diagnosis, Prognosis and Treatment of Canine Mammary Tumors. Brazilian Journal of Veterinary Pathology, 4(2): 153-180.


• Janeklang, S., Nakaew, A., Vaeteewoottacharn, K., Seubwai, W., Boonsiri, P., Kismali, G., et al. (2014). In vitro and in vivo antitumor activity of tiliacorinine in human cholangiocarcinoma. Asian Pacific Journal of Cancer Prevention, 15(17): 7473-7478.


• Kim, K.H., Moon, E., Kim, S.Y., Choi, S.U. and Lee, K.R. (2012). Lignan constituents of Tilia amurensis and their biological evaluation on antitumor and anti - inflammatory activities. Food and Chemical Toxicology, 50(10): 3680-3686.


• Marrassini, C., Anesini, C. and Ferraro, G. (2011): HPLC fingerprint of a flower extract of Tilia x viridis and correlation with antiproliferative and antioxidant activity. Phytotherapy Research, 25(10): 1466-1471.


• Pan, D., Boon-Unge, K., Govitrapong, P. and Zhou, J. (2011). Emetine regulates the alternative splicing of caspase 9 in tumor cells. Oncology Letters, 2(6):1309-1312.


• Pecorino, L. (2012). Molecular Biology of Cancer. 3rd ed., UK: Oxford University Press, pp: 10-11.


• Ruberte, J., Sautet, J.Y., Gine, J.M., Lopez, C. and Rodriguez, A. (1990). Topography of lymphatic drainage from mammary glands of the bitch. Anatomia, Histologia, Embryologia, 19(4): 358-347.


• Sefidabi, R., Mortazavi, P. and Hoseni, S. (2016). Antiproliferative effect of berberine on canine mammary gland cancer cell culture. Biomedical Report, 6(1): 95-98.


• Slatter, D. (2003). Text Book of Small Animal Surgery.3rd ed., Elsevier Science, pp: 349-350.


• Yayalai, Y., Celik, I. and Bati B. (2014). Hepatoprotective and antioxidant activity of Linden (Tilia platyphyllos L.) infusion against ethanol-induced oxidative stress in rats. The Journal of Membrane Biology, 247(2): 181-188.


• Zargari, A. (1989). Medicinal Plants. 7th ed., Iran: Tehran University Press, pp: 399. [In Persian]