Inhibition of proliferation and apoptosis of human colon cancer cell sw480 cell line induced by the co-culture supernatant of Toxoplasma gondii
10.3969/j.issn.1002-2694.2010.03.010
- VernacularTitle:刚地弓形虫细胞培养上清对人结肠癌细胞sw480增殖与凋亡的影响
- Author:
Jian GAO
;
Bin YE
;
Weihua WU
- Publication Type:Journal Article
- Keywords:
Toxoplasma gondii;
human colon cancer cell line sw480;
culture;
apoptosis
- From:
Chinese Journal of Zoonoses
2010;(3):229-234
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the inhibitive effect of the co-culture supernatant of Toxoplasma gondii on the proliferation of human colon cancer cell line sw480 and/or its induced apoptosis and necrosis in vitro,the co-culture model of sw480 cells (1x10~6) was established with different number of Toxoplasma gondii (the concentration of tachyzoites was 2×10~6,4×10~6,8×10~6,16×10~6 respectively),and the co-culture supernatants were gathered 72 hours later.The sw480 cells were treated for different hours with different models of the co-culture supernatant.The parasitism and proliferation of Toxoplasma gondii tachyzoites in sw480 cells was observed by Giemsa staining,and the growth inhibition rates of these cells were investigated by the CKK-8 method.Meanwhile,the apoptosis and necrosis of sw480 cells were detected with transmission electron microscopy (TEM) and fluorescence microscopy with Hochest 33258 staining to observe the morphological changes of sw480 cells.Agarose electrophoresis was used to detect the DNA change of sw480 cells,and flow cytometric analysis was used for investigation of rates of apoptosis and necrosis of these cells stained with Annexin-v-FITC/PI.It was found that the parasitism and proliferation of Toxoplasma gondii tachyzoites in sw480 cells could be observed;and as demonstrated by the CKK-8 method,the inhibitive effect of the co-culture supernatant depended upon the treatment time,and a maximum inhibition ratio of 44.55% was detected at 48 hours.The karyopyknosis and karyorrhexis in the treated sw480 cells were observed under the fluorescence microscope;and the cellular necrosis were observed under TEM.In addition,the DNA fragments in the treated cells were revealed by agarose electrophoresis.In the flow cytometric analysis,a maximum prophase apoptosis rate of 11.54% was detected at 48 hours after treatment with the supernatants,while the rate of prophase apoptosis decreased,and the rates of advanced stage/necrosis increased markedly at the treatment time beyond 48 hours.It is concluded that the co-culture supernatant of Toxoplasma gondii and human colon cancer cell sw480 cell line can induce inhibition of proliferation,apoptosis and necrosis of this cell line in vitro.
