EXPLORING THE ANTI-CERVICAL CANCER POTENTIAL OF CASSIA ANGUSTIFOLIA: INSIGHTS FROM HELA CELL LINE, AND MOLECULAR DOCKING
Main Article Content
Keywords
Cassia angustifolia, HeLa cell line, anticancer phytocompounds, gene expression, molecular docking
Abstract
Cassia angustifolia (CA: senna makkai) is a medicinal plant that belongs to Saudi Arabia and Egypt and is widely cultivated in Pakistan. This study was designed for the in vitro anticancer assessment of Cassia extracts on cervical (HeLa) cancer and noncancer human embryonic kidney (HEK-293T) cell lines. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) and crystal violet assays were performed to determine the cytotoxicity of extracts. For the mRNA expression analysis of p53 and Bcl2, the SYBR Green-based Real-time PCR was performed. Molecular interaction analysis and binding affinity were explored between target proteins (TP53, BCL2, EGFR, and HER2) and compounds through computational approaches. GCMS was used to identify the phytocompounds with anticancer potential using ethanol (ECA) and n-hexane (HCA) solvent systems. GCMS analysis showed the presence of bioactive phytochemicals in both CA extracts. These extracts demonstrated dose-dependent growth inhibition of tumor cells with an IC50 value of 50 and 76 µg/mL. Significant downregulation of TP53 (tumor suppressor) and upregulation of the BCL-2 (antiapoptotic) proteins was observed after treatment with CV extracts. Phenol, 2-methyl-5-(1,2,2-trimethylcyclopentyl)-, (S)- was observed as the most potential bioactive compound towards TP53 (-7.5) (kcal/mol) protein, followed by Isophytol (-6.9). This work highlights cancer-target proteins involved in proliferation and drug-like compounds for cervical cancer treatment. Based on all these findings, it is concluded that ECA extract has promising anticancer potential and might be valued for the advancement of novel plant-derived drugs for treating and managing cervical cancer.
References
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28. Nawaz, A., Jamal, A., Arif, A., and Parveen, Z. (2021). In vitro cytotoxic potential of Solanum nigrum against human cancer cell lines. Saudi Journal of Biological Sciences, 28(8), 4786-4792.
29. Nelson, V. K., Sahoo, N. K., Sahu, M., Sudhan, H. H., Pullaiah, C. P., and Muralikrishna, K. S. (2020). In vitro anticancer activity of Eclipta alba whole plant extract on colon cancer cell HCT-116. BMC complementary medicine and therapies, 20, 1-8.
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44. Tulashie, S. K., Adjei, F., Abraham, J., and Addo, E. (2021). Potential of neem extracts as natural insecticide against fall armyworm (Spodoptera frugiperda (JE Smith)(Lepidoptera: Noctuidae). Case Studies in Chemical and Environmental Engineering, 4, 100130.
45. Umar HI, Siraj B, Ajayi A, Jimoh TO, and Chukwuemeka PO (2021). Molecular docking studies of some selected gallic acid derivatives against five non-structural proteins of novel coronavirus. Journal of Genetic Engineering and Biotechnology, 19(1): https://doi.org/10.1186/s43141-021-00120-7
46. Umesh HR, Ramesh KV, and Devaraju KS. Molecular docking studies of phytochemicals against trehalose-6-phosphate phosphatases of pathogenic microbes. Beni-Suef University Journal of Basic and Applied Sciences, 2020;9(1):1–14. doi: 10.1186/s43088-019-0028-6
47. Zhu, S., Jiao, W., Xu, Y., Hou, L., Li, H., Shao, J., and Kong, D. (2021). Palmitic acid inhibits prostate cancer cell proliferation and metastasis by suppressing the PI3K/Akt pathway. Life Sciences, 286: 120046.
2. Ahmed, S. I., Hayat, M. Q., Tahir, M., Mansoor, Q., Ismail, M., Keck, K., and Bates, R. B. (2016). Pharmacologically active flavonoids from the anticancer, antioxidant and antimicrobial extracts of Cassia angustifolia Vahl. BioMed Central complementary and alternative medicine, 16(1): 1-9.
3. Aung, T. N., Qu, Z., Kortschak, R. D., and Adelson, D. L. (2017). Understanding the effectiveness of natural compound mixtures in cancer through their molecular mode of action. International journal of molecular sciences, 18(3), 656.
4. Baeshen NA, Albeshri AO, Baeshen NN, Attar R, Karkashan A, Abbas B, Baeshen MN (2022). In silico screening of some compounds derived from the desert medicinal plant Rhazya stricta for the potential treatment of COVID-19. Sci. Rep 12(1): 11120. https://doi.org/10.1038/s41598-022-15288-2
5. Cang, S., Iragavarapu, C., Savooji, J., Song, Y., and Liu, D. (2015). ABT-199 (venetoclax) and BCL-2 inhibitors in clinical development. Journal of hematology & oncology, 8(1), 1-8.
6. Çankaya N, Azarkan SY, Tanış E (2022). The molecular interaction of human anti-apoptotic proteins and in silico ADMET, drug-likeness and toxicity computation of N-cyclohexylmethacry lamide. Drug Chem. Toxicol 45(5), 1963-1970. https://doi.org/10.1080/01480545.2021.1894711
7. Entezari, M., and Sheikhan, S. (2018). Evaluation of Bcl2 gene expression in MCF-7 Human Breast Cancer Cells under treatment of Centaurea Behen extract and Cisplatin. Archives of Advances in Biosciences, 9(4): 11-16.
8. Ezzat, S. M., Shouman, S. A., Elkhoely, A., Attia, Y. M., Elsesy, M. S., El Senousy, A. S., & El Tanbouly, N. (2018). Anticancer potentiality of lignan-rich fraction of six Flaxseed cultivars. Scientific Reports, 8(1), 544.
9. Federico, C., Sun, J., Muz, B., Alhallak, K., Cosper, P. F., Muhammad, N., and Azab, A. K. (2021). Localized delivery of cisplatin to cervical cancer improves its therapeutic efficacy and minimizes its side effect profile. International Journal of Radiation Oncology* Biology* Physics, 109(5), 1483-1494.
10. Gorji-Bahri, G., Moradtabrizi, N., and Hashemi, A. (2021). Uncovering the stability status of the reputed reference genes in breast and hepatic cancer cell lines. Plos one, 16(11): e0259669.
11. Graur, F., Furcea, L., Mois, E., Biliuta, A., Rozs, A. T., Negrean, V., and Al Hajjar, N. (2016). Analysis of p53 Protein Expression in Hepatocellular Carcinoma. Journal of Gastrointestinal and Liver Diseases, 25(3).
12. He, M., Xia, L., and Li, J. (2021). Potential mechanisms of plant-derived natural products in the treatment of cervical cancer. Biomolecules, 11(10), 1539.
13. Islam, M. T., Ali, E. S., Uddin, S. J., Shaw, S., Islam, M. A., Ahmed, M. I., and Atanasov, A. G. (2018). Phytol: A review of biomedical activities. Food and chemical toxicology, 121: 82-94.
14. Kalsoom, A., Altaf, A., Jilani, M. I., Sattar, H., Maqbool, T., and Muhammad, T. (202). In vitro antiproliferative potential of Cassia angustifolia extracts on HepG2 cells to combat liver cancer. International Journal of Applied and Experimental Biology.
15. Kaur, S., Pandit, K., Chandel, M., and Kaur, S. (2020). Antiproliferative and apoptogenic effects of Cassia fistula L. n-hexane fraction against human cervical cancer (HeLa) cells. Environmental Science and Pollution Research, 27(25): 32017-32033.
16. Kikuchi, H. Yuan, B. Hu, X and Okazaki, M. Chemopreventive and anticancer activity of flavonoids and its possibility for clinical use by combining with conventional chemotherapeutic agents. American Journal of Cancer Research, 2019, 9, 1517–1535.
17. Kiran, A., Altaf, A., Sarwar, M., Malik, A., Maqbool, T., and Ali, Q. (2023). Phytochemical profiling and cytotoxic potential of Arnebia nobilis root extracts against hepatocellular carcinoma using in-vitro and in-silico approaches. Scientific Reports, 13(1), 11376.
18. Kumari, S., Singh, A., Sangal, R., and Sharma, N. R. (2021). KAP study on cervical cancer and human papillomavirus vaccine acceptability among adolescent girls of Eastern UP: a cross sectional study. International Journal of Reproduction, Contraception, Obstetrics and Gynecology, 10(5), 2031-2036.
19. Lipinski, C.A., Lombardo, F., Dominy, B.W., Feeney, P.J., 1997. Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Advance Drug Delivery Reviews, 23 (1–3), 3–25.
20. Mariamenatu, A. H., and Abdu, E. M. (2021). Overconsumption of omega-6 polyunsaturated fatty acids (PUFAs) versus deficiency of omega-3 PUFAs in modern-day diets: the disturbing factor for their “balanced antagonistic metabolic functions” in the human body. Journal of Lipids, 1-15.
21. Masuku, N. P., and Lebelo, S. L. (2019). Investigation of the effects of Kigelia africana (Lam.) Benth. Extracts on TM3 Leydig cells. Asian Journal of Pharmacuetical and Clinical Research, 12(10): 87-92.
22. Mbugua, R. W., Njagi, E. M., Ngule, C. M., and Mwitari, P. (2019). Gene expression mediated antiproliferative potential and safety of selected medicinal plants against cancerous and normal cells. BioRxiv, 578948.
23. Mendie LE, and Hemalatha S (2022). Molecular docking of phytochemicals targeting GFRs as therapeutic sites for cancer: An in silico study. Applied Biochemistry and Biotechnology, 1-17 (194): 215–231. https://doi.org/10.1007/s12010-021--03791-7
24. Mohamad Rosdi, M. N., Mohd Arif, S., Abu Bakar, M. H., Razali, S. A., Mohamed Zulkifli, R., and Ya’akob, H. (2018). Molecular docking studies of bioactive compounds from Annona muricata Linn as potential inhibitors for Bcl-2, Bcl-w and Mcl-1 antiapoptotic proteins. Apoptosis, 23, 27-40.
25. Momtazi-Borojeni, A. A., Ghasemi, F., Hesari, A., Majeed, M., Caraglia, M., and Sahebkar, A. (2018). Anticancer and radio-sensitizing effects of curcumin in nasopharyngeal carcinoma. Current pharmaceutical design, 24(19), 2121-2128.
26. Morris, J. L., Gillet, G., Prudent, J., and Popgeorgiev, N. (2021). Bcl-2 family of proteins in the control of mitochondrial calcium signalling: an old chap with new roles. International Journal of Molecular Sciences, 22(7), 3730.
27. Nakamura, M., Obata, T., Daikoku, T., and Fujiwara, H. (2019). The association and significance of p53 in gynecologic cancers: the potential of targeted therapy. International Journal of Molecular Sciences, 20(21), 5482
28. Nawaz, A., Jamal, A., Arif, A., and Parveen, Z. (2021). In vitro cytotoxic potential of Solanum nigrum against human cancer cell lines. Saudi Journal of Biological Sciences, 28(8), 4786-4792.
29. Nelson, V. K., Sahoo, N. K., Sahu, M., Sudhan, H. H., Pullaiah, C. P., and Muralikrishna, K. S. (2020). In vitro anticancer activity of Eclipta alba whole plant extract on colon cancer cell HCT-116. BMC complementary medicine and therapies, 20, 1-8.
30. Ouyang, L.; Luo, Y.; Tian, M.; Zhang, S.-Y.; Lu, R.; Wang, J.-H.; Kasimu, R.; Li, X. Plant natural products: From traditional compounds to new emerging drugs in cancer therapy. Cell Proliferation. 2014, 47, 506–515.
31. Park, S. H., Kim, M., Lee, S., Jung, W., and Kim, B. (2021). Therapeutic potential of natural products in treatment of cervical cancer: a review. Nutrients, 13(1), 154.
32. Pillaiyar, T., Meenakshisundaram, S., and Manickam, M. (2020). Recent discovery and development of inhibitors targeting coronaviruses. Drug discovery today, 25(4), 668-688.
33. Purkiewicz, A., Czaplicki, S., and Pietrzak-Fiećko, R. (2022). The Occurrence of Squalene in Human Milk and Infant Formula. International Journal of Environmental Research and Public Health, 19(19): 12928.
34. Ravindranath PA, Forli S, Goodsell DS, Olson AJ, Sanner MF (2015). AutoDockFR: advances in protein-ligand docking with explicitly specified binding site flexibility. PLoS Computional Biology, 11(12): e1004586. https://doi.org/10.1371/journal.pcbi.1004586
35. Sadia, H., Bhinder, M. A., Irshad, A., Zahid, B., Ahmed, R., Ashiq, S., and Akbar, A. (2020). Determination of expression profile of p53 gene in different grades of breast cancer tissues by real time PCR. African Health Sciences, 20(3): 1273-1282.
36. Salam, H. S., Tawfik, M. M., Elnagar, M. R., Mohammed, H. A., Zarka, M. A., and Awad, N. S. (2022). Potential Apoptotic Activities of Hylocereus undatus Peel and Pulp Extracts in MCF-7 and Caco-2 Cancer Cell Lines. Plants, 11(17): 2192.
37. Sattar H, Firyal S, Awan AR, Habib-ur-Rehman, Tayyab M, Hasni SM, Ali MM, Saeed S, Mehmood T, Aqib IA, Khan HM, Wasim M (2021). Genetic Association of Polymorphism and Relative mRNA Expression of Tumor Necrosis Factor-Alpha Gene in Mastitis in Sahiwal Cows. International Journal of Agricultural and Biological Engineering, 25:701‒708. https://doi.org/10.17957/IJAB/15.1719
38. Shivanika C, Kumar D, Ragunathan V, Tiwari P, Sumitha A (2020). Molecular docking, validation, dynamics simulations, and pharmacokinetic prediction of natural compounds against the SARS-CoV-2 main-protease. Journal of Biomolecular Structure and Dynamics. https://doi.org/10.1080/07391102. 2020.1815584
39. Singh, R., Letai, A., and Sarosiek, K. (2019). Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nature reviews Molecular cell biology, 20(3), 175-193.
40. Solowey, E., Lichtenstein, M., Sallon, S., Paavilainen, H., Solowey, E., and Lorberboum-Galski, H. (2014). Evaluating medicinal plants for anticancer activity. The Scientific World Journal, 2014.
41. Sultana T, Mitra AK, and Das S (2022). Evaluation of the anticancer potential of Excoecaria agallocha (L.) leaf extract on human cervical cancer (SiHa) cell line and assessing the underlying mechanism of action. Future journal of Pharmaceutical Sciences, 8(1): https://doi.org/10.1186/s43094-021-00389-y
42. Sun, C, Brown, A. J., Jhingran, A., Frumovitz, M., Ramondetta, L., and Bodurka, D. C. (2014). Patient preferences for side effects associated with cervical cancer treatment. International journal of gynecological cancer: official journal of the International Gynecological Cancer Society, 24(6), 1077.
43. Sung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A., and Bray, F. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians, 71(3), 209-249.
44. Tulashie, S. K., Adjei, F., Abraham, J., and Addo, E. (2021). Potential of neem extracts as natural insecticide against fall armyworm (Spodoptera frugiperda (JE Smith)(Lepidoptera: Noctuidae). Case Studies in Chemical and Environmental Engineering, 4, 100130.
45. Umar HI, Siraj B, Ajayi A, Jimoh TO, and Chukwuemeka PO (2021). Molecular docking studies of some selected gallic acid derivatives against five non-structural proteins of novel coronavirus. Journal of Genetic Engineering and Biotechnology, 19(1): https://doi.org/10.1186/s43141-021-00120-7
46. Umesh HR, Ramesh KV, and Devaraju KS. Molecular docking studies of phytochemicals against trehalose-6-phosphate phosphatases of pathogenic microbes. Beni-Suef University Journal of Basic and Applied Sciences, 2020;9(1):1–14. doi: 10.1186/s43088-019-0028-6
47. Zhu, S., Jiao, W., Xu, Y., Hou, L., Li, H., Shao, J., and Kong, D. (2021). Palmitic acid inhibits prostate cancer cell proliferation and metastasis by suppressing the PI3K/Akt pathway. Life Sciences, 286: 120046.
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Feb 22, 2024
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Aasia Kalsoom, Awais Altaf, Tahir Maqbool, Saira Aftab, Huma Sattar, Muhammad Sajjad, & Asia Kiran. (2024). EXPLORING THE ANTI-CERVICAL CANCER POTENTIAL OF CASSIA ANGUSTIFOLIA: INSIGHTS FROM HELA CELL LINE, AND MOLECULAR DOCKING. Journal of Population Therapeutics and Clinical Pharmacology, 31(2), 2174–2195. https://doi.org/10.53555/jptcp.v31i2.4594
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Author Biographies
Aasia Kalsoom
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan
Awais Altaf
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan
Tahir Maqbool
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan
Saira Aftab
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan
Huma Sattar
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan
Muhammad Sajjad
School of Biological Sciences, Punjab University, Lahore, Pakistan
Asia Kiran
Institute of Molecular Biology and Biotechnology (IMBB/CRiMM), The University of Lahore, Lahore, Pakistan