The expression of p53 and hsp70 proteins after treatment with Annona muricata Linn leaf for activating apoptotic and lead to homeostasis program of Raji cells

Okid Parama Astirin, Adi Prayitno, Anif Nur Artanti, Meutia Srikandi Fitria, Dyah Ayu Witianingsih, Dwimei Ayudewandari Pranatami, Suhartono Taat Putra

Abstract


Purpose: Organic extracts of plant Annonaceae enhances apoptosis in animal cells and get the drives to reach a new homeostasis. The incidence rate of nasopharyngeal cancer in Indonesia is quite high. Protein 53Kd (p53) play a role in apoptosis process, being heat shock protein 70 (hsp70) play a role in homeostasis. The aim of this research is to identify the apoptotic effects of Annona muricata Linn leaf toward Raji cells by observing the p53 and hsp70 expression.

Methods: Apoptotic assay was performed in 24 wells micro-culture plate. Raji cells were prepared as 2 × 104 cells in 100 ml RPMI media per well. Roswell Park Memorial Institute (RPMI) medium was created and solvent was controlled with Dimethyl Sulfoxide (DMSO) solvent 0.25. Apoptotic test was performed by calculating trypan-blue-dye exclution. The cells were then grown in micro-culture plate with media plus extract non-lethal concentration of partition and fractionation of Annona muricata Linn leaf. The sampling was performed for 24 hours. The number of living cells was calculated in each of these well and incubation time were determined. Immunohistochemical staining was done to identify the expression of p53 and hsp70.

Results: The results showed that Raji cells treated with partition of Annona muricata Linn leaf in ethyl acetate solvent 133.00 % resulted in higher apoptosis. Another results showed that Raji cells treated with fractionation Annona muricata Linn leaf in ethyl acetate solvent 103.20 % resulted in higher apoptosis. The expression of p53 after treatment with fractionation Annona muricata Linn leaf was higher than before while hsp70 expression after treatment with fractionation Annona muricata Linn leaf was lower.

Conclusion: The conclusion is the higher the dose of Annona muricata Linn the higher the p53 expression thereby activates apoptosis process The higher dose of Anonna muricata Linn also leads to lower hsp70 expression indicating stable homeostasis of Raji cells.

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Cite this article as: Astirin OP, Prayitno A, Artanti AN, Fitria MS, Witianingsih DA, Pranatami DA, Putra ST. The expression of p53 and hsp70 proteins after treatment with Annona muricata Linn leaf for activating apoptotic and lead to homeostasis program of Raji cells. Int J Cancer Ther Oncol 2014; 2(2):02028. DOI: 10.14319/ijcto.0202.8


Keywords


Annona Muricata Linn Leaf; p53; hsp70; Apoptosis; Homeostasis; Raji Cell

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References


Hildesheim A, West S, DeVeyra E, et al. Herbal medicine use, Epstein-Barr virus, and risk of nasopharyngeal carcinoma. Cancer Res 1992; 52:3048-51.

Astirin O, Artanti A, Fitria M, et al. Annonaa muricata Linn Leaf Induce Apoptosis in Cancer Cause Virus. Journal of Cancer Therapy 2013; 4:1244-50.

Yoshizaki T, Ito M, Murono S, et al. Current understanding and management of nasopharyngeal carcinoma. Auris Nasus Larynx 2012; 39: 137-44.

Yu MC, Yuan JM. Epidemiology of nasopharyngeal carcinoma. Semin Cancer Biol 2002; 12:421-9.

Chang ET, Adami HO. The enigmatic epidemiology of nasopharyngeal carcinoma. Cancer Epidemiol Biomarkers Prev 2006; 15:1765-77.

Cao SM, Simons MJ, Qian CN. The prevalence and prevention of nasopharyngeal carcinoma in China. Chin J Cancer 2011; 30:114-9.

Smardova J, Pavlova S, Svitakova M, et al. Analysis of P53 Status in Human Cell Line Using a Functional Assay in Yeast: Detection of New no Sense p53 Mutattion in Codon 124. Oncol Rep 2005; 14:901-7.

Hicks SD, Lewis L, Ritchie J, et al. Evaluation of cell proliferation, apoptosis, and DNA-repair genes as potential biomarkers for ethanol-induced CNS alterations. BMC Neurosci 2012; 13:128.

Adham M, Kurniawan AN, Muhtadi AI, et al. Nasopharyngeal carcinoma in Indonesia: epidemiology, incidence, signs, and symptoms at presentation. Chin J Cancer 2012; 31:185-96.

Selye, H. Stress in Health and Disease. Batterworths, Boston and London . Ann Intern Med 1977; 87:799.

Vogel EH, Castro ME, Solar PA, Soto FA. Enhancement of Pavlovian conditioned immunosuppression in rats. Acta Neurobiol Exp (Wars) 2007; 67:71-81.

Tao Q, Chan AT. Nasopharyngeal carcinoma: molecular pathogenesis and therapeutic developments. Expert Rev Mol Med 2007; 9:1-24.

Burgos JS. Involvement of the Epstein-Barr virus in the nasopharyngeal carcinoma pathogenesis. Med Oncol 2005; 22:113-21.

Cregan SP, MacLaurin JG, Craig CG, et al. Bax-dependent caspase-3 activation is a key determinant in p53-induced apoptosis in neurons. J Neurosci 1999; 19:7860-9.

Bagatell R, Whitesell L. Altered Hsp90 function in cancer: a unique therapeutic opportunity. Mol Cancer Ther 2004; 3:1021-30.

Seoane JM, Varela-Centelles PI, Ramirez JR, et al. Heat shock proteins (HSP70 and HSP27) as markers of epithelial dysplasia in oral leukoplakia. Am J Dermatopathol 2006; 28:417-22.

Mese H, Sasaki A, Nakayama S, et al. Prognostic significance of heat shock protein 27 (HSP27) in patients with oral squamous cell carcinoma. Oncol Rep 2002; 9:341-4.

Macario AJ, Conway de Macario E. Sick chaperones, cellular stress, and disease. N Engl J Med 2005; 353:1489-501.

Morishima N. Control of cell fate by Hsp70: more than an evanescent meeting. J Biochem 2005; 137:449-53.

Freshney R Ian, Culture of Animal Cells: A manual of Basic Techniques 4th Ed., Wiley-Lies, Canada. 2000.

Mursyidi ,Achmad. Statistika Farmasi dan Biologi, Ghalia, Indonesia, Jakarta. 1985.

Prayitno A, Asnar E, Astirin O, Putra S. The Expression of CD8+ and MHC-I in Cervical Cancer with HPV Infection. Journal of Cancer Therapy 2013; 4:15-8.

Rohman A, Man YB, Riyanto S. Authentication analysis of red fruit (Pandanus Conoideus Lam) oil using FTIR spectroscopy in combination with chemometrics. Phytochem Anal 2011; 22:462-7.

Hamizah S, Roslida AH, Fezah O, et al. Chemopreventive potential of Annona muricata L leaves on chemically-induced skin papillomagenesis in mice. Asian Pac J Cancer Prev 2012; 13:2533-9.

Shan T, Ma Q, Guo K, et al. Xanthones from mangosteen extracts as natural chemopreventive agents: potential anticancer drugs. Curr Mol Med 2011; 11:666-77.

Tan W, Lu J, Huang M, et al. Anti-cancer natural products isolated from chinese medicinal herbs. Chin Med 2011; 6:27.

Teppo L, Pukkala E, Saxen E. Multiple cancer--an epidemiologic exercise in Finland. J Natl Cancer Inst 1985; 75:207-17.

Zhu J, Okumura H, Ohtake S, et al. Arsenic trioxide induces apoptosis in leukemia/lymphoma cell lines via the CD95/CD95L system. Oncol Rep 2003; 10:705-9.

Alberts B, Johnson A, Lewis J, et al. Molecular biology of the cell, 4th edition Garland Science, New York 2002.

Prayitno A, Darmawan R, Yuliadi I, Mudigdo A.The Expression of p53, Rb dan c-myc proteins in cervical cancer by immunohistochemistry stain. Journal of Biological Diversity 2005; 6.

Prayitno A. Cervical cancer with human papilloma virus and Epstein Barr virus positive. J Carcinog 2006; 5:13.

Prayitno A, Asnar E, Astirin O, et al. Heat Shock Protein 40 (Hsp40) and Hsp70 Protein Expression in Oral Squamous Cell Carcinoma (OSCC). Journal of Cancer Therapy 2013; 4:734-41.

Astirin O.P, Implantasi pada Rattus norvegicus setelah perlakuan dengan Actinomycin D. Thesis, Faculty of Pasca Sarjana, University of Gajah Mada, Yogyakarta, 1991.

Macario AJ, Lange M, Ahring BK, Conway de Macario E. Stress genes and proteins in the archaea. Microbiol Mol Biol Rev 1999; 63:923-67.

Horst M, Oppliger W, Rospert S, et al. Sequential action of two Hsp70 complexes during protein import into mitochondria. EMBO J 1997;16:1842-9.

Neupert W. Protein import into mitochondria. Annu Rev Biochem 1997; 66:863-917.

Ryan MT, Naylor DJ, Hoj PB, Clark MS, et al. The role of molecular chaperones in mitochondrial protein import and folding. Int Rev Cytol 1997; 174:127-93.

Jiang Z, Chen RY, Chen Y, Yu DQ. Donnaienin, a New Acetogenin Bearing a Hydroxylated Tetrahydrofuran Ring. J Nat Prod 1998; 61:86-8.

Rieser MJ, Gu ZM, Fang XP, et al. Five novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona muricata. J Nat Prod 1996; 59:100-8.




DOI: http://dx.doi.org/10.14319/ijcto.0202.8

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