Genotoxic effects of gossypol acetic acid on cultured murine erythroleukemia cells

Abstract

Gossypol acetic acid, a male anti-fertility drug, was evaluated for its effects on cell multiplication, chromosomes, scheduled and unscheduled DNA synthesis, and the surface ultrastructure in cultured murine erythroleukemia cells (clone 6A11A). Gossypol treatments inhibited cell multiplication at 10 and 20 micrograms/ml concentrations and this inhibitory effect increased with elevated dosage and prolonged treatment. Gossypol significantly depressed the mitotic index but did not alter chromosome numbers or increase the frequency of chromosomal structural abnormalities. Cell fraction techniques revealed that gossypol induced a negative effect on cellular DNA synthesis at concentrations as low as 3.3 micrograms/ml after 24 hr of treatment. The number of cells undergoing DNA synthesis decreased with increasing dosages and durations of drug exposure. An unscheduled DNA synthesis assay (UDS) found gossypol to be an active UDS-inducing agent at certain dose levels and treatment times, as measured by increase in net nuclear gain and percentage of UDS cells (ANOVA, Bonferroni test, P less than 0.05). A scanning electron microscope study revealed that 10 micrograms/ml treatment with gossypol caused changes in mouse erythroleukemia cell surface ultrastructure characterized by general balding and the appearance of holes, often after 48 hr of treatment.

Differential cytotoxic effects of azadirachtin on Spodoptera frugiperda and mouse cultured cells

“…SF-9 cells were cultured aseptically in Grace's medium containing 10% fetal calf serum and maintained at 27 • C in a non-humidified incubator. The cell doubling time for SF-9 under optimum conditions ranges between 20-24 h. The fast growing mouse erythroleukemia cells (MEL-GM 86) grow in suspension and double every 12-14 h. The cells were purchased from the Human Cell Repository (Camden, NJ) and maintained in Dulbecco's Modified Eagle's Medium containing 10% fetal calf serum at 37 • C in a humidified incubator with 7.5% CO 2 in air (Majumdar et al, 1991).…”

“…Centrifuged cells were fixed in 3:1 methanol/glacial acetic acid and stained with Giemsa stain. (Majumdar et al, 1991(Majumdar et al, , 1997. Slides were made in triplicate and 500 random cells were counted per slide to calculate the average percent of enlarged and damaged cells for the control and treatment groups.…”

Silica Gel Chromatographic Methods for Identification, Isolation and Purification of Gossypol Acetic Acids Prepared in Lab

“…Its toxicity was investigated on different cancer cell lines such as Her2 positive breast cancer cells [15]. It is able to cause up-regulation of somatostatin receptors 2 and 5 in DU-145 prostate cancer cells [16], and cultured murine erythroleukemia cells [17]. It can inhibit the growth of colorectal cancer and hepatocellular carcinoma [18] and the like.…”