COMPARATIVE EVALUATION OF ANTI-PROLIFERATIVE ACTIVITY OF SOLANUM NIGRUM METHANOLIC AND AQUEOUS EXTRACT ON HELA, SIHA AND C33A CELLS

摘要

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. Keywords: Cervical cancer cell line, Solanum nigrum , DNA laddering, Caspase-independent apoptosis, Annexin V, Autophagy.