Prodigiosin, the red pigment of Serratia marcescens, shows cytotoxic effects and apoptosis induction in HT-29 and T47D cancer cell lines.

In this study, a red pigment of Serratia marcescens PTCC 1111 was purified and identified for antiproliferative activities in HT-29 and T47D cancer cell lines. (1)H-NMR spectroscopy and LC/MS analysis confirmed prodigiosin structure. The antiproliferative effects of prodigiosin were determined by employing the MTT assay. The changes in cell cycle pattern were studied with 4',6-diamidino-2-phenylindole (DAPI) reagent using flow cytometry assay, and Annexin V-PI method was used for apoptotic analysis. Results of MTT assay showed that HT-29 cells were more sensitive to prodigiosin than T47D cells. Prodigiosin-treated HT-29 cells showed increase in S phase and decrease in G2/M, but treated T47D cells showed cell cycle pattern relatively similar to Roswell Park Memorial Institute medium (RPMI). Apoptotic effect of prodigiosin was higher than doxorubicin in HT-29 cells. The data reported here indicate that prodigiosin is a promising antineoplastic agent that triggers apoptosis in different cancer cell lines.

Comparative cellular and molecular analysis of cytotoxicity and apoptosis induction by doxorubicin and Baneh in human breast cancer T47D cells.

It is now widely accepted that dietary phytochemicals inhibit cancer progression and enhance the effects of conventional chemotherapy. In this report, we comparatively studied the cellular and molecular aspects of apoptosis induction by the methanolic extract of Baneh fruit skin in comparison to Doxorubicin (Dox), a well-known anticancer drug, in human breast cancer T47D cells. The MTT assay was used to determine the antiproliferative effects. The flow cytometric and microscopic analyses were done to evaluate the apoptosis induction. Furthermore, western blot analyses have been done to study the role of key molecular players of apoptosis including caspase 3 and PARP. The Baneh extract showed strong antiproliferative activity against T47D cells in a dose- and time-dependent manner that was comparable to and even stronger than Dox in certain concentrations. Analysis of Baneh-treated cells by flow cytometry and fluorescence microscopy indicated strong apoptosis induction and nuclear morphological alterations similar to or greater than Dox. Finally, molecular analysis of apoptosis by western blotting proved activation of caspase 3 followed by poly ADP ribose polymerase (PARP) cleavage more efficiently in Baneh than in Dox treated cancer cells. These findings indicate that Baneh extract contains phytochemicals which act as inhibitor of cell proliferation and inducer of apoptosis in human breast cancer T47D cells that makes it a potentially good candidate for new anticancer drug development.

Preparation and evaluation of poly (caprolactone fumarate) nanoparticles containing doxorubicin HCI.

BACKGROUND AND THE PURPOSE OF THE STUDY: Biodegradable Poly(caprolactone fumarate) (PCLF) has been used as bioresorbable sutures. In this study, doxorubicin HCl (Dox) loaded PCLF nanoparticles were prepared and characterized. MATERIAL AND METHODS: PCLFs were synthesized by polycondensation of PCL diols (Mws of 530, 1250 and 2000) with fumaryl chloride. The degradation of PCLF in NaOH, water and phosphate buffer saline (PBS), was determined in terms of changes in Mw. Nanoparticles (NPs) were prepared by two methods. In microemulsion polymerization method, dichloromethane containing PCLF and photoinitiator were combined with the water containing surfactants and then the mixture was placed under light for crosslinking. In nanoprecipitation method, the organic solvent containing PCLF was poured into the stirring water. The effect of several variables including concentration of PCLF, polyvinyl alcohol (PVA), Dox and Trypan blue (Trb) and the Mw of PCLF and PVA on NP size and loading were evaluated. RESULT: PCLF 530, 1250 and 2000 in PBS or water were not degraded over 28 days. Nanoprecipitaion method gave spherical (revealed by SEM images) stable NPs of about 225 with narrow size distribution and a zeta potential of -43 mV. The size of NP increased significantly by increase in Mw or concentration of PCLF. Although PVA was not necessary for formation of NPs, but it decreased with NP size. Dox loading and EE were 2.5-6.8% and 15-20%, respectively. Increasing the drug concentration increased the drug loading (DL) and NP size. The entrapment efficiency (EE) for Trb ranged from 1% for PCLF530 to 6% for PCLF2000. An increase in PCLF concentration resulted in an increase in EE. Dox and Trb release showed a burst followed by 80% and 78% release during 3 and 4 days respectively. CONCLUSION: PCLF possessed suitable characteristics for preparation of nanoparticulate drug delivery system such as desired NP size, stability and degradation time. Although PCLF530 NPs were the smallest, but their DL were lower than PCLF1250 and 2000 NPs.

Molecular analysis of MEN1 expression in MCF7, T47D and MDA-MB 468 breast cancer cell lines treated with adriamycin using RT-PCR and immunocytochemistry.

BACKGROUND AND THE PURPOSE OF THE STUDY: MEN1 is an important tumor suppressor gene that encodes a nuclear protein called menin. Recent data suggest that interactions between menin and other proteins have important roles in control of the cell cycle and apoptosis. In addition, estrogen receptor (ER), an important prognostic factor is differentially expressed in breast cancer cells. In this study the MEN1 gene and protein expression in MCF7, T47D and MDA-MB-468 breast cancer cell lines with different ER status following exposure to adriamycin (ADR) was investigated. MATERIALS AND METHODS: Cytotoxicity of ADR on these cell lines was determined using MTT assay. The mRNA and protein levels were analyzed in tested cell lines using RT-PCR and immunocytochemistry (ICC) assays, respectively. RESULTS: ADR cytotoxicity was highest on MDA-MB-468 and lowest on MCF7 cells. MEN1 mRNA showed significant decrease after ADR exposure only in the MDA-MB-468 cell line. Menin protein expression was higher in MDA-MB-468 and lower in MCF7 cells. CONCLUSION: Differential molecular responses to adriamycin were observed in cancer cell lines. Molecular data also suggest that MEN1 as a new biomarker can be used in combination with current biomarkers for prediction of response to chemotherapy.