刚地弓形虫培养上清抑制THP-1细胞株增殖

Culture supernatant of Toxoplasma gondii inhibits the proliferation of human acute monocytic leukemia cell line THP-1

目的提取弓形虫体外细胞共培养上清,研究上清对人急性单核细胞白血病细胞株THP-1增殖及细胞周期的影响。方法取对数生长期的THP-1细胞(浓度为5×105/mL)分别接种于不同细胞培养瓶,对照组加入含10%胎牛血清RPMI-1640,实验组加入相同体积不同数量(2×107、4×107和8×107/mL)弓形虫速殖子培养上清孵育不同时间后,MTT法检测THP-1细胞增殖率;流式细胞仪检测细胞周期;Western blot检测细胞核转录因子NF-κB/P65与周期蛋白CYCLIND1表达或活性。结果弓形虫培养上清呈时间剂量依赖性抑制THP-1细胞株增殖,使细胞周期在G0/G1期产生阻滞;8×107/mL数量组的THP-1细胞48 h抑制率可达(23.71±0.56)%,G0/M1期细胞比例达(58.53±1.03)%,较对照组比较有明显差异(P<0.05),且处理组THP-1细胞株的NF-κB/P65、cyclin D1蛋白表达量下降。结论刚地弓形虫培养上清能够通过NF-κB信号途径下调cyclin D1蛋白表达引起人急性单核细胞白血病细胞株THP-1 G0/G1期阻滞。

Objective The find how to get culture supernatants of Toxoplasma gondii and investigate the inhibitive effects of culture supernatants of Toxoplasma gondii on the proliferation and on the cell cycle of Human acute monocytic leukemia cell line THP-1 in vitro.Methods THP-1 cells（concentration of 5×104/mL） were harvested in different cell culture plates.The cells were treated for different hours with different concentrations of Toxoplasma gondii culture supernatant.Growth inhibition rate was tested with the MTT method;Cell cycle was checked by Flow Cytometer.Western blot was used to detect the level of NF-κB/P65 and cyclin D1 of cells.Results The culture supernatants of Toxoplasma gondii inhibited proliferation of THP-1 cells in a time-dose dependent manner.Cell cycle was significantly stopped at G0/G1phase by the culture supernatants with FCM technology.The culture supernatant of Toxoplasma gondii reduced the expressions of gene NF-κB、CYCLIND1 of THP-1 cells.Conclusions The culture supernatant of Toxoplasma gondii may inhibit THP-1 cell and arrest the cell cycle of THP-1 cells at G0/ G1 phase mainly by regulating the expression of gene NF-κB、cyclin D1.