Cytotoxicity and cell death mechanisms against cancer cell lines elicited by the extracts of physalis minima L.

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dc.contributor.authorKheng Leong, Ooi
dc.date.accessioned2015-07-30T04:43:00Z
dc.date.available2015-07-30T04:43:00Z
dc.date.issued2009
dc.description.abstractHerbal therapy is fast becoming an important alternative medicine for cancer treatment. Therefore, the aims of this study were to determine cytotoxicity of some medicinal herbs and to investigate cell death mechanism elicited by the most potent extract as well as its fractions. A total of 20 extracts form four anticancer plants (Pereskia grandifo/ia, Vernonia cinerea, Elephantopus scaber and Physalis minima) were screened against three different cancer cell lines. Physalis minima (Leletup-direct translation from Malay) chloroform extract was shown to exhibit remarkable cytotoxic effects on NCI-H23 (human lung adenocarcinoma), T-470 (human breast carcinoma) and Caov-3 (human ovarian carcinoma) cell lines, with EC50 derived at 2.80f.!g/ml, 3.80f.!g/ml and 5.10f.lg/ml, respectively. Different cell death mechanisms exerted by this extract on different cell lines. It was found that DNA fragmentation level of the extract-treated NCI-H23 and T-470 cells was higher than Caov-3 cells. Acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in all cell lines. Ultrastructural analysis and annexin V staining also demonstrated the presence of apoptotic programmed cell death in the extract-treated cell lines. Furthermore, the appearance of non-apoptotic (vacu?lar) morphology was observed in some treated Caov-3 cells and in minority of treated NCI-H23 and T-470 cells. Trypan blue exclusion assay ruled out necrosis as the main cause of death. Thus, cell death mechanism elicited by the Physalis minima chloroform extract appeared to be a mixture of programmed cell death in Caov-3 cells and a major apoptotic cell death in both NCI-H23 and T-470 cells. Fractionation of Physalis minima chloroform extract revealed 16 different fractions. Only some of the fractions exhibited cytotoxic activities against T-470 cells, with Fll being the most potent, exhibiting the lowest EC50 (3.60f.!g/ml). Fractions FlO, Fll and F13 exhibited typical DNA fragmentation associated with apoptosis in T-470 cells. Trypan blue exclusion assay demonstrated that necrosis did not play a major role in eliciting T-47D cell death of these fractions. Further investigations revealed both fraction F11 and chloroform extract induced major apoptotic cell death in T-47D cells, which was based on majority of weakly diffused annexin V stained cells and significant regulation of c-myc, caspase-3 and p53 mRNA expression levels. Meanwhile, a considerable number of T-47D cells treated with fractions FlO and Fl3 were stained with annexin V and propidium iodide, which further suggested the presence of a mixture programmed cell death. Physalins B, F and K were identified as major constituents in fraction F 11. Combination of these physalin compounds in the extract and fraction may have contributed to the cytotoxic activities and programmed celt death of T-47D cells. These findings warrant further research on Physalis minima plant for use in cancer therapy due to its apoptosis- and autophagy-dependent anticancer effect on cancer cell lines.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/1016
dc.language.isoenen_US
dc.subjectCell deathen_US
dc.subjectCancer cell linesen_US
dc.subjectPhysalis minima l.en_US
dc.titleCytotoxicity and cell death mechanisms against cancer cell lines elicited by the extracts of physalis minima L.en_US
dc.typeThesisen_US
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