Thymoquinone could increase the efficacy of tamoxifen induced apoptosis in human breast cancer cells: an in vitro study

Objective: Thymoquinone [TQ], as the main component of Nigella Sativa plant, shows anticancer properties. This study was aimed to evaluate the combined effect of TQ and Tamoxifen [TAM] on viability and apoptosis of human breast cancer cell lines

Materials and Methods: In this experimental study, estrogen positive MCF-7 and estrogen negative MDA-MB-231 human breast cancer cell lines were induced by TAM [2 microM] or different doses of TQ [50, 75, 100, 150 microM], individually or in combination. Cell viability and apoptosis were investigated by MTT assay and TdT-mediated deoxy-uracil nick end labeling [TUNEL] assay; Acridine orange [AO]/Ethidium bromide [EB] staining respectively. Data were analyzed by one way ANOVA and P<0.05 was considered significant

Results: In 24 hours treatment, TAM and all doses of TQ, solely or in combination, significantly reduced cell viability of both cell lines, except in MCF-7 cells treated with 50 microM TQ, and MDA-MB-231 cells treated with 50 or 75 microM TQ [P<0.01]. After 48 hours treatment, cell viability of both cell lines was reduced in all treated groups [P<0.05]. Remarkable apoptotic index was observed in combination treatment of MCF-7 or MDA-MB-231 cell lines with TAM and TQ [P<0.001]

Conclusion: The synergistic effect of TQ and TAM on human breast cancer cell lines showed cell viability reduction as well as apoptosis induction, independent to estrogen

Hydrostatic pressure affects in vitro maturation of oocytes and follicles and increases granulosa cell death

This study examines the effects of hydrostatic pressure on in vitro maturation [IVM] of oocytes derived from in vitro grown follicles. In this experimental study, preantral follicles were isolated from 12-day-old female NMRI mice. Each follicle was cultured individually in Alpha Minimal Essential Medium [alpha-MEM] under mineral oil for 12 days. Then, follicles were induced for IVM and divided into two groups, control and experiment. In the experiment group follicles were subjected to 20 mmHg pressure for 30 minutes and cultured for 24-48 hours. We assessed for viability and IVM of the oocytes. The percentage of apoptosis in cumulus cells was determined by the TUNEL assay. A comparison between groups was made using the student's t test. The percentage of metaphase II oocytes [MII] increased in hydrostatic pressure-treated follicles compared to controls [p<0.05]. Cumulus cell viability reduced in hydrostatic pressure-treated follicles compared to controls [p<0.05]. Exposure of follicles to pressure increased apoptosis in cumulus cells compared to controls [p<0.05]. Hydrostatic pressure, by inducing apoptosis in cumulus cells, participates in the cumulus oocyte coupled relationship with oocyte maturation

Antiproliferatory Effects of Crab Shell Extract on Breast Cancer Cell Line (MCF7) / 한국유방암학회지

PURPOSE: Breast cancer is the most common type of cancer in women. Despite various pharmacological developments, the identification of new therapies is still required for treating breast cancer. Crab is often recommended as a traditional medicine for cancer. This study aimed to determine the in vitro effect of a hydroalcoholic crab shell extract on a breast cancer cell line. METHODS: In this experimental study, MCF7 breast cancer cell line was used. Crab shell was powdered and a hydroalcoholic (70degrees ethanol) extract was prepared. Five concentrations (100, 200, 400, 800, and 1,000 microg/mL) were added to the cells for three periods, 24, 48, and 72 hours. The viability of the cells were evaluated using trypan blue and 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Cell apoptosis was determined using the terminal deoxynucleotidyl transferase dUTP nick end labeling method. Nitric oxide (NO) level was assessed using the Griess method. Data were analyzed using analysis of variance, and p<0.05 was considered significant. RESULTS: Cell viability decreased depending on dose and time, and was significantly different in the groups that were treated with 400, 800, and 1,000 microg/mL doses compared to that in the control group (p<0.001). Increasing the dose significantly increased apoptosis (p<0.001). NO secretion from MCF7 cells significantly decreased in response to different concentrations of the extract in a dose- and time-dependent manner (p<0.050). CONCLUSION: The crab shell extract inhibited the proliferation of MCF7 cells by increasing apoptosis and decreasing NO production.

Hydrostatic pressure improves in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries

Cryopreservation has limited successes and in-vitro maturation is used to improve its results. Hydrostatic pressure [HP] plays an important role in follicular development. This study was designed to examine the effects of HP on in-vitro maturation of oocytes and cell death in cumulus cells derived from vitrified-warmed mouse ovaries. Preovulatory follicles were harvested from non-vitrified and vitrified-warmed 6-8 week-old female NMRI mouse ovaries and randomly assigned to following groups: non-vitrified [control], non-vitrified with HP exposure [treatment I], vitrified-warmed [treatment II] and vitrified-warmed with HP exposure [treatment III]. The follicles of treatments I and III were subjected to HP [20 mmHg] for 30 min and after that all groups were cultured for 24h and assessed for in-vitro maturation of oocytes. The viability and apoptosis of cumulus cells and oocytes were assessed using supravital nuclear staining and TUNEL assay, respectively. Oocytes harvested follicles in both control and treatment II had a significantly lower percentage of metaphase II oocytes [MII] than the treatment I and III [23.5 +/- 3.1, 15.03 +/- 4.6 and 32.7 +/- 3.2, 25.5 +/- 4.6; respectively] [p<0.05]. Viability of the cumulus cells reduced in treatment I, II and III [83.4, 83.3 and 77.7%] compared to control [86.9%], [p<0.05]. The apoptotic index in cumulus and oocyte complexes in treatments I and III [10.7 +/- 0.8 and 15.3 +/- 0.8] was higher than in control and treatment II [6.7 +/- 0.5 and 9.7 +/- 0.5] [p<0.05]. These results demonstrate that HP had a mild effect on cell death incidence in cumulus cells without any effect on oocyte. However, it can be used as a mechanical force to improve in-vitro maturation of oocytes derived from vitrified-warmed mouse ovaries