Effect of Lycium depressum Root on Bax , Bcl-2 Gene Expressions and Evaluation of Caspase 3 and 6 Activity on Uterine Cervical Cancer cells line (Hela)

Subject Areas : Journal of Animal Biology

Yasamin Khani ¹ , Tahereh Naji ² , Soheila Zamanlui Benisi ³ , Saeid Mohamadi Motamed ⁴

1 - Department of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences Unit, Islamic Azad University, Tehran, Iran
2 - Department of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences Unit, Islamic Azad University, Tehran, Iran
3 - Department of Stem Cells and Cell Therapy, Research Institute of Tissue Engineering and Regenerative Medicine, Central Tehran Branch, Islamic Azad University, Tehran, Iran
4 - Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences Unit, Islamic Azad University, Tehran, Iran

Received: 2023-01-31 Accepted : 2023-04-21 Published : 2023-11-22

Keywords: Gene expression, cervical cancer, Apoptosis, Lycium depressum,

Abstract :

Cervical cancer is the third cause of death caused by cancer in women. After breast cancer, cervical cancer is the most common cancer in women. Today, due to the numerous side effects of chemotherapy and radiation therapy that are caused to the patient as a result of their use, as well as the resistance of cancer cells to common treatments, researchers have turned to new drugs with greater effectiveness and less toxicity. The aim of the present study was to evaluate the effect of Lycium depressum root on the expression of Bcl2 and Bax genes and to evaluate the activity of caspases 3 and 6 in cervical cancer cells (Hela). In this study, firstly, the extract of Lycium depressum root was obtained by soaking method, then cervical cancer cells were prepared and cultured, in order to calculate the optimal effective dose of the extract, the MTT technique was used. The concentrations of 25 and 50 mg/ml were chosen as optimal doses. Then the expression of Bcl2 and Bax genes was evaluated by Real-time PCR technique. Next, in order to evaluate the activity of caspases 3 and 6 in effective doses of the extract, cervical cancer cells (Hela) were affected and protein expression was measured using the immunocytochemistry (ICC) technique. The results of this research showed that the roots of the Turkmen oleander plant can cause changes in the expression of important Bcl2 and Bax genes in the path of apoptosis induction. In addition, it increases the protein expression of caspase 3 and 6 in cancer cells. Therefore, it can be hoped that in additional research on the extract of Lycium depressum root, it will be considered a candidate for the treatment of complementary medicine.

References:

1. Abdullaev F., 2001. Plant-derived agent’s cancer. Journal of Pharmacology, 15(3):345-354.


2. Ahmadi R., Rahimi S., Ehteshamzad N., 2017. The effect of hydroalcoholic Ziziphus spina-christi leaf extract on viability of breast cancer cell line (MCF7) and evaluation of Bax and Bcl2 genes expression level. Kashan university of medicinal Journal, 21(5):407-413.


3. Baharlou R., Atashzar MR., Vasmehjani AA., Rahimi E., Khoshmirsafa M., Seif F., et al., 2016. Reduced levels of T-helper 17-associated cytokines in the serum of patients with breast cancer: indicators for following the course of disease. Central European Journal Immunology, 41(1):78.


4. Bedoui S., Herold MJ., Strasser A., 2020. Emerging connectivity of programmed cell death pathways and its physiological implications. Nature Reviews Molecular Cell Biology, 21(11):675-695.


5. Cowling V., Downward J., 2002. Caspase-6 is the direct activator of caspase-8 in the cytochrome c-induced apoptosis pathway: absolute requirement for removal of caspase-6 prodomain. Cell Death and Differentiation, 9(10):1046-1056.


6. Cox JT., Castle PE., Kitchener HC., 2006. Achievements and limitations of cervical cytology screening. Vaccine, 24: S63-S70.


7. Fitzmaurice C., Allen C., Barber RM., Barregard L., Bhutta ZA., Brenner H. 2017. Global burden of disease cancer collaboration. global, regional, and national cancer Incidence, mortality, years of life lost, years lived with disability, and disability-adjusted Life-years for 32 cancer groups, 1990 to 2015: A Systematic Analysis for the Global Burden of Disease Study. Jama Oncology, 3(4):524-548.


8. Ghorbani A., Ghorbanihesari T., Mortazavian M., 2012. Effect of aqueous extract – alcohol b and its fractions brtksyr cervical cancer cells. The Iranian Journal of Obstetrics, Gynecology and Infertility, 15(22):9-16.


9. Guo Y., Srinivasula SM., Druilhe A., Fernandes-Alnemri T., Alnemri ES., 2002. Caspase-2 induces apoptosis by releasing proapoptotic proteins from mitochondria. Journal of Biological Chemistry, 277(16):13430-13437.


10. Harlev E., Nevo E., Lansky EP., Lansky S., Bishayee A., 2012. Anticancer attributes of desert plants: a Anticancer Drugs, 23:255-271.


11. Horn LC., Raptis G., Fischer U., 2002. Familial cancer history in patients with carcinoma of the cervix uteri. European Journal Obstetrics and Gynecology and Reproductive Biology, 101(1):54-57.


12. Horner-Johnson W., Dobbertin K., Andresen EM., Iezzoni LI., 2014. Breast and cervical cancer screening disparities associated with disability severity. Women’s Health, 24(1):e147-e153.


13. Jafari M., Moradi Y., Khodadost M., Sekhavati , Anabad HA., Mansori K., et al., 2015. Trend of the esophageal cancer incidence in Iran. International Journal of Travel Medicine Global Health, 3(2):127-131.


14. Lazhiry A., Vlak J., 1993. Herb plants. Translated to Persian by: Zaman S. Tehran: Ghoghnoos Pub.


15. Leece P., Kendall C., Touchie C., Pottie K., Angel JB., Jaffey J.,2010. Cervical cancer screening among HIV-positive women. Retrospective cohort study from a tertiary care HIV clinic. Canadian Family Physician, 56(12):e425-431.


16. Moshtaghi , Esmaeilzadeh Bahabadi S., Mazaheri MA., Najafi SH., Dahmardeh Ghalenoo F. 2016. Increasing of caspase3 gene expression in Mcf7 breast cancer cell line by Nigella sativa hydro-alcoholic extract. Shahid Sadoughi University Journals, 24(1):1-1.


17. Saxena A., Viswanathan S., Moshynska O., Tandon P., Sankaran K., Sheridan DP., 2004. Mcl‐1 and Bcl‐2/Bax ratio are associated with treatment response but not with Rai stage in B‐cell chronic lymphocytic leukemia. American Journal of Hematology, 75(1):22-33.


18. Sonnenschein C., Soto AM., 2008. Theories of carcinogenesis: an emerging perspective. Seminars in Cancer Biology, 18(5):372-377.


19. Thompson CB., 1995. Apoptosis in the pathogenesis and treatment of disease. Science, 267(5203):1456-1462.


20. Vogelstein B., Kinzler KW., 2004. Cancer genes and the pathways they control. Nature Medicine,10(8):789-799.


21. Wang S-J., Zheng C-J., Peng C., Zhang H., Jiang Y-P., Han T., et al., 2013. Plants and cervical cancer: an overview. Expert opinion on Investigational Drugs, 22(9):1133-1156.


22. Xiao X., Ren W., Zhang N., Bing T., Liu X., Zhao Z., Shangguan D. 2019. Comparative study of the chemical constituents and bioactivities of the extracts from fruits, leaves and root barks of Lycium barbarum. Molecules, 24(8):1585.


23. Xiong L., Deng N., Zheng B., Li T., Liu RH., 2021. Goji berry (Lycium spp.) extracts exhibit antiproliferative activity via modulating cell cycle arrest, cell apoptosis, and the p53 signaling pathway. Food and Function, 12(14):6513-6525.


24. Yao X., Peng Y., Xu LJ., Li L., Wu QL., Xiao PG., 2011. Phytochemical and biological studies of Lycium medicinal plants. Chemistry and Biodiversity, 8(6):976-1010.


25. Zhu C.P., Zhang S.H. 2013. Lycium barbarum polysaccharide inhibits the proliferation of HeLa cells by inducing apoptosis. Journal of the Science of Food and Agriculture, 93:149-560.