COMPARATIVE EVALUATION OF ANTI-PROLIFERATIVE ACTIVITY OF SOLANUM NIGRUM METHANOLIC AND AQUEOUS EXTRACT ON HELA, SIHA AND C33A CELLS
Keywords:
Cervical cancer cell line, Solanum nigrum, DNA laddering, Caspase-independent apoptosis, Annexin V, AutophagyAbstract
Objective: Comparative evaluation of anti-proliferative activities of aqueous (SNA) and methanolic extract (SNM) of Solanum nigrum was studied on HeLa, SiHa (HPV-infected) and C33A (non-HPV) cell lines.
Methods: Cellular and nuclear morphology of treated cells were studied by phase contrast and fluorescence microscopy respectively. Cell cycle analysis was carried out by flow cytometry. DNA fragmentation assay and Annexin-V assay were conducted to study apoptosis induction. Expression of apoptotic proteins – Caspase and Bax were studied at translational level.
Results: SNM treated cells showed chromatin condensation, nuclear distortion and fragmentation as well as DNA laddering, whereas extensive cytoplasmic vacuolation was observed on treatment with SNA. In case of treatment with SNM, higher percentage of cells in sub G0 phase was observed in HeLa (40.83%) and C33A (44.72%) as compared to SiHa (11.95%). Flowcytometric assays showed higher percentage of late and early apoptotic cells in HeLa (58.5%) as compared to SiHa (31.3%) and C33A (41.5%) cell lines. Down regulation of Caspase and Bax was observed in all three cell lines treated with SNM and SNA. Cleaved Caspase was absent in case of all the three cell lines.
Conclusion: Absence of cleaved Caspase suggests that cell death by SNM might be taking place by Caspase –independent apoptotic pathway, while occurrence of cytoplasmic vacuoles in SNA treated cells suggest that cell death might be taking place by autophagic pathway.
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References
Kritikar KR, Basu BD, Blatter E. Indian medicinal plants. 2nd Ed. (2), Allahabad, Lalit Mohan Basu Publications; 1935.
Jainu M, Devi CS. Antiulcerogenic and ulcer healing effects of Solanum nigrum (L.) on experimental ulcer models: Possible mechanism for the inhibition of acid formation. J Ethnopharmacol 2006;104:156-63.
Jain R, Sharma A, Gupta S, Sarethy IP, Gabrani R. Solanum nigrum: Current perspective on therapeutic properties. Altern Med Rev 2011;16:78-85.
Patel S, Gheewala N, Suthar A, Shah A. In-vitro cytotoxicity activity of Solanum nigrum extract against HeLa cell line and vero cell line. Int J Pharm Pharm Sci 2009;1(1):38-46.
Wang W, Lu DP. An in vitro study of cytotoxic and antineoplastic effect of Solanum nigrum extracts on U266. Beijing Da Xue Xue Bao 2005;37:240-4.
Gabrani R, Jain R, Sharma A, Sarethy IP, Dang S, Gupta S. Antiproliferative Effect of Solanum nigrum on Human Leukemic Cell Lines. Indian J Pharm Sci 2012;74(5):451-3.
Tai CJ, Wang CK, Tai CJ, Lin JF, Lin CS, Jian JJ, Chang JJ, et al. Aqueous Extract of Solanum nigrum Leaves Induces Autophagy and Enhances Cytotoxicity of Cisplatin, Doxorubicin, Docetaxel, and 5-Fluorouracil in Human Colorectal Carcinoma Cells. J Evid Based Complementary Altern Med 2013. Available from: http: //dx.doi.org/ID.1155/2013/514719.
Vallikad E. Cervical cancer: The Indian perspective. Int J Gynaecol Obstet 2006;95(1):S215-S233.
Kurkure AP, Yeole BB. Social inequalities in cancer with special reference to South Asian countries. Asian Pac J Cancer Prev 2006;7(1):36-40.
WHO/ICO Information Centre on HPV and Cervical Cancer (HPV Information Centre). Human papillomavirus and related cancers in India. Summary Report 2010. Available from: www.hpvcentre.net/statistics/reports/IND.pdf.
Dohun Pyeon, Michael A Newton, Paul F Lambert, Johan A den Boon, Srikumar Sengupta, Carmen J Marsit, et al. Fundamental differences in cell cycle deregulation in human papillomavirus–positive and human papillomavirus–negative head/neck and cervical cancers. Cancer Res 2007;67:4605-19.
Li LY, Luo X, Wang X. Endonuclease G is an apoptotic DNase when released from mitochondria. Nat 2001;412:95–9.
Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE. Molecular characterization of mitochondrial apoptosis-inducing factor. Nat 1999;397:441–6.
Kroemer G, Galluzzi L, Vandenabeele P, Abrams J, Alnemri ES. Classification of cell death: recommendations of the nomenclature committee on cell death 2009. Cell Death Differ 2009;16:3–11.