INDUCTION OF AUTOPHAGY IN HUMAN CERVICAL CANCER CELL LINE (SIHA) BY CHAETOMORPHA LINUM (MULLER) KÃœTZ.
DOI:
https://doi.org/10.22159/ijpps.2018v10i6.24593Keywords:
Chaetomorpha, SiHa cell line, Autophagy, Western blot analysis, LC3B, GCMSAbstract
Objective: Algae is a rich source of various metabolites. Use of algae in traditional medicine was reported from very early days. The potential anticancerous activity of the algae was also reported by recent researches. In the present study, cell death mechanism induced by green alga Chaetomorpha linum (Muller) Kütz. was studied on human cervical cancer cell line, SiHa (Human papilloma virus-HPV 16+ve).
Methods: Cells were treated with chloroform fraction of the Chaetomorpha linum (CLC). DNA fragmentation, cell proliferation, nuclear morphology, localization of autophagosomes and expression of relevant proteins were studied. Cell cycle and mitochondrial membrane potential (MMP) were assessed by fluorescence assisted cell sorting (FACS). Gas chromatography liquid spectrometry (GCMS) analysis of CLC was done to detect the algal compounds.
Results: Preliminary experiments confirmed the absence of DNA fragmentation and altered nuclear morphology in the treated cells. In the Acridine Orange stained cells, the presence of autophagosomes was observed by both microscopy and FACS analysis. Decreased MMP was observed in the treated sets. Wound healing assay showed the inhibitory activity of CLC on SiHa cells. Increased expression of autophagic proteins, viz; LC3BII, p62/SQSTM, Beclin-1 and decreased expression of S6 were observed in the treated sets; indicating that autophagic cell death was induced. Preliminary qualitative chemical study and GCMS analysis detected the presence of saponins, glycosides, alkaloids, phenols, steroids and fatty acids in CLC.
Conclusion: All these findings clearly suggested that the chloroform fraction of the algae was responsible for induction of apoptosis followed by autophagic cell death in SiHa and can serve as a potential candidate for anticancer drug development in future.
Downloads
References
Faulkner DJ. Marine natural products. Nat Prod Rep 2001;18:1–49.
Al-Saif SSA, Abdel-Raouf N, El-Wazanani HA, Aref IA. Antibacterial substances from marine algae isolated from Jeddah coast of Red sea, Saudi Arabia. Saudi J Biol Sci 2014;21:57-64.
Kolanjinathan K, Ganesh P, Govindarajan M. Antibacterial activity of ethanol extract of seaweeds against fish bacterial pathogens. Eur Rev Med Pharmacol Sci 2009;13:173-7.
Princely S, Dhanaraju MD. Assessment of phytochemical constituent in vitro antimicrobial and antioxidant potential of Ulva extracts from Vishakhapatnam coast. Asian J Pharm Clin Res 2017;10:87-95.
Sheu JH, Huang SY, Wang GH, Duh CY. Study on cytotoxic oxygenated desmosterols isolated from the red alga Galaxaura marginata. J Nat Prod 1997;60:900-3.
Kwon MJ, Nam TJ. A polysaccharide of the marine alga Capsosiphon fulvescens induces apoptosis in AGS gastric cancer cells via an IGF-IR-mediated PI3K/Akt pathway. Cell Biol Int 2007;31:768-75.
Fischel JL, Lemee R, Formento P, Caldani C, Moll JL, Pesando D. Cell growth inhibitory effects of caulerpenyne, a sesquiterpenoid from the marine alga Caulerpa taxifolia. Anticancer Res 1995;15:2155-60.
Barbier P, Guise S, Huitorel P, Amade P, Pesando D, Briand C. Caulerpenyne from Caulerpa taxifolia has an antiproliferative activity on tumor cell line SK-N-SH and modifies the microtubule network. Life Sci 2001;70:415–29.
Harada H, Noro T, Kamei Y. Selective antitumor activity in vitro from marine algae from Japan Coasts. Biol Pharm Bull 1997;20:541-6.
Ahmad A, Usman H, Natsir H, Karim A. Isolation and characterization of a bioactive protein from green algae Halimeda macrobola acting as antioxidant and anticancer agent. Am J Biomed Life Sci 2014;2:134-40.
Moo-Puc R, Robledo D, Freile-Pelegrin Y. In vitro cytotoxic and antiproliferative activities of marine macroalgae from Yucatán, Mexico. Cienc Mar 2009;35:345–58.
Paul S, Kundu R. Antiproliferative activity of methanolic extracts from two green algae, Enteromorpha intestinalis and Rizoclonium riparium on HeLa cells. DARU J Pharm Sci 2013;21:72.
Forman D, Bray F, Brewster DH, Gombe Mbalawa C, Kohler B, Ferlay J, et al. Cancer incidence in five continents, Vol. X (electronic version) Lyon, IARC; 2013. Available from: http://ci5.iarc.fr [Last accessed on 01 Nov 2013]
Majumder I, Chatterjee S, Kundu R. A study of the antiproliferative potentiality of some major macroalgae from sundarbans on cervical cancer cell line. J Algal Biomass Utln 2015;6:212.
Kang H, Alvarado AS. Flow cytometry methods for the study of cell-cycle parameters of planarian stem cells. Dev Dyn 2009;238:1111–7.
Hu H, Ahn HN, Yang X, Lee YS, Kang KS. Ganoderma lucidum extract induces cell cycle arrest and Apoptosis in mcf-7 human breast cancer cell. Int J Cancer 2002;102:250–3.
Pozo Guisado E, Alvarez Barrientos A, Mulero Navarro S, Santiago Josefat B, Fernandez Salguero PM. The antiproliferative activity of resveratrol results in apoptosis in MCF-7 but not in MDA-MB-231 human breast cancer cells: cell-specific alteration of the cell cycle. Biochem Pharmacol 2002;64:1375-86.
Hamori E, Arndt Jovin DJ, Grimwade BG, Jovin TM. Selection of viable cells with known DNA content. Cytomet 1980;1:1980.
Loken MR. Simultaneous quantitation of hoechst 33342 and immunofluorescence on viable cells using a fluorescence activated cell sorter. Cytomet 1980;1:136–42.
Trease GE, Evans WC. Pharmacognosy. 11th ed. London: MacMillan Publishers Ltd.; 1989. p. 60-75.
Feng Z, Zhang H, Levine AJ, Jin S. The coordinate regulation of the p53 and mTOR pathways in cells. Proc Natl Acad Sci USA 2005;102:8204-9.
Glick D, Barth S, Macleod KF. Autophagy: cellular and molecular mechanisms. J Pathol 2010;221:3-12.
Tang J, Di J, Cao H, Bai J, Zheng J. p53-mediated autophagic regulation: A prospective strategy for cancer therapy. Cancer Lett 2015;63:101-7.
Kim YS, Shin JH, Bae MK. Autophagy activity in pulmonary metastatic tumor tissues from colorectal cancer: a pilot study. Yonsei Med J 2014;55:1484-8.
Marks N, Berg MJ, Guidotti A, Saito M. Activation of caspase-3 and apoptosis in cerebellar granule cells. J Neurosci Res 1998;52:326-41.
Rajkumar S, Jebanesan A. Mosquitocidal activities of octacosane from Moschosma polystachyum Linn. (lamiaceae). J Ethnopharmacol 2004;90:87-9.
Ahsan T, Chen J, Zhao X, Irfan M, Wu Y. Extraction and identification of bioactive compounds (eicosane and dibutyl phthalate) produced by Streptomyces strain KX852460 for the biological control of Rhizoctonia solaniAG-3 strain KX852461 to control target spot disease in tobacco leaf. AMB Express 2017;7:54.
Ansari KF, Lal C. Synthesis and evaluation of some new benzimidazole derivatives as potential antimicrobial agents. Eur J Med Chem 2009;44:2294-9.
Kristina S, Marijeta M, Katja E, Ivan S, Magdalena G, Grace KP, et al. Synthesis, the antiviral and antitumor activity of 2-substituted-5-amidino-benzimidazoles. Bioorg Med Chem 2007;15:4419.
Sondhi SM, Rani M, Sigh J, Roy P, Agrawal SK, Saxena AK. Solvent-free synthesis, anti-inflammatory and anticancer activity evaluation of tricyclic and tetracyclic benzimidazole derivatives. Bioorg Med Chem Lett 2010;20:2306.
Mu YM, Yanase T, Nishi Y, Tanaka A, Saito M, Nawata H, et al. Saturated FFAs, palmitic acid and stearic acid, induce apoptosis in human granulosa cells. Endrocrinology 2001;142:3590-7.
Ulloth JE, Casiano CA, De Leon M. Palmitic and stearic fatty acids induce caspase-dependent and-independent cell death in nerve growth factor differentiated PC12 cells. J Neurochem 2003;84:655-68.
Published
How to Cite
Issue
Section
