Apoptotic and necrotic effects of pectic acid on rat pituitary GH3/B6 tumor cells

ABSTRACT

Pectin is composed of complex polysaccharides that can inhibit cancer metastasis and proliferation with no evidence of toxicity. In the present study, the apoptotic and necrotic effects of pectic acid [PA] on the rat pituitary GH3/B6 tumor cells has been investigated. GH3/B6 cells were cultured in the Ham's F12 medium enriched with 15% horse serum and 2.5% fetal bovine serum for 3 days. Then, they were treated by various amounts of PA in different periods [6, 24 and 48 hours]. Bromocriptine was used as positive control and the cell viability was detected by MTT test. The nuclear morphology of cells was explored by florescent stains including acridine orange [AO]/ethidium bromide [EB]. In addition, percentages of apoptotic and necrotic cells were studied with triphosphate nick-end labeling [TUNEL] assay, cell cycle analysis and propidium iodide [PI] staining. Long-term incubation with PA results in increased cell death and DNA damage as detected by MTT assay and AO/EB staining. TUNEL assay showed that PA [100 micro g/ml to 1 mg/ml] could induce apoptosis in a dose-dependent manner, while higher concentrations of PA [2.5 and 5 mg/ml] induced necrosis which was confirmed by PI staining. Furthermore, cell cycle analysis indicated that PA induced sub G1 events, and DNA fragmentation was also correlated with the number of the apoptotic cells. It can be concluded that PA is responsible for apoptosis in the rat pituitary tumor cells. Therefore, one may suggest that this group of polysaccharides can be used in treatment of pituitary tumors