In vitro cytotoxicity assay of Fucoidan extracted from Turbinaria conoides against cancer cell lines MCF7, A549, and normal cell line L929

Abstract The main aim of the study is to quantify the cytotoxic property of the Fucoidan extracted from the Turbinaria conoides using the MTT assay with the standard fucose. Fucoidan was extracted using the soaked water method and it was determined using the HPLC procedure the obtained Test sample Fucoidan extracted from the Turbinaria conoides and standard fucose was subjected to the cytotoxicity assay against the MCF7 Human breast cancer cell line, A549 lung cancer cell line, and L929 normal mouse fibroblast cell line. From the results it was found that the Test sample showed good IC50 value for MCF7 cell line then A549 with an increasing concentration 24 hours incubation at 37°C The IC50 for MCF7 was 115.21 µg/ml and A549 396.46µg/ml and the Fucoidan extract was checked for its cytotoxicity against the normal mouse fibroblast cell line L929, Fucoidan was found non-lethal to the L929 mouse fibroblast normal cell line. Standard fucose also gave a significant result towards MCF7 and against the L929. This indicates that the Fucoidan extracted from Tubinaria conoides shows better anticancer potential in it. Hence its application can be further extended in the pharmacological fields.

Diallyl Disulfide from Garlic Induces Apoptosis through a Caspase Dependent Pathway in Human Breast Cancer Cell Line, MCF-7

PURPOSE: Diallyl disulfide (DADS), an organosulfur compound in garlic, has been reported to be effective in inhibiting the growth of several human tumor cell lines. The aim of this study was to determine whether DADS induced growth inhibition in MCF-7 breast cancer cell lines and to understand the molecular mechanism by which DADS acts. METHODS: MCF-7 cell lines were incubated with various concentrations of DADS for various time intervals and the cytotoxicity was determined by MTT assay. We examined the changes of intracellular proteins related to apoptosis, such as bcl-2, bax and PARP in cells treated with DADS. To study the expression level of bcl-2 and bax, which serve as modulators of apoptosis, we performed RT-PCR and western blot analysis. RESULTS: MCF-7 cells treated with DADS led to the suppression of viability and proliferation in both a time and concentration dependent manner. Microscopic observation revealed typical features of apoptosis in the DADS-treated cells, further verified in nuclear DAPI staining. Flow cytometry analysis with FITC-annexinV and propidium iodide (PI) demonstrated that the apoptotic cell population with AnnexinV /PI increased dramatically from ~0.8% to ~75% after 24h exposure to 500nM DADS in MCF-7 cells. Cellcycle analysis demonstrated that the number of apoptotic cells increased with the increasing time of the DADS treatment. Additionally, thermore, we investigated the effects of DADS on apoptosis related gene expression in MCF-7 cells. PARP cleavage was markedly increased in the DADS treated cells with time. This result indicated that DADS induced the caspase-dependent apoptotic pathway. We also found down-regulation of bcl-2, however no significant change of Bax expression was observed after DADS treatment. CONCLUSION: Taken together, these results indicate that DADS induces apoptosis by activating a caspase pathway involving the activation of Bcl-2 but not of Bax. Our findings suggest chemotherapeutic potentials of DADS in human breast cancer.

The Effect of Fentanyl on Proliferation and Cell Cycle of Human Breast Carcinoma Cell Line MCF-7 / 中国医师杂志

Objective To study the effects and its mechanism of fentanyl(Fen) on the proliferation and cell cycle of human breast carcinoma line MCF-7. Methods MCF-7 cells were cultured in the medium with Fen, naloxone(Nx) or both the medicines at different concentration for different time. MTT method was employed to evaluate the level of the cell proliferation. The distribution of the cell cycle was detected with the flow cytometry (FCM). The expression levels of p53 and p21/WAF1 in the cells were determined by SP immunocytochemical staining method. Results Fen at≥0.1?mol/L concentration inhibited MCF-7 cells proliferation in dose- and time-dependent manners, and its IC 50 for 72h was 0.81?0.02 ?mol/L. However, the antiprolifeative effect of Fen was not antagonized by Nx. Fen significantly enhanced the ratio of G 0/G 1 phase MCF-7 cells, and decreased the proliferation index of MCF-7 cells in dose-dependent manner. Fen also upregulated the expression of p53 and p21/WAF1 in MCF-7 cells. Conclusion The data suggested that the inhibitory effect of Fen on MCF-7 cell growth might be mediated by blocking cell cycle progression from G 1 to S phase, and upregulating the expression of p53 and p21/WAF1.