弓形虫感染小鼠血清对小鼠B16黑色素瘤细胞增殖和凋亡的影响

Proliferation and apoptosis of B16 melanoma cells in the sera of mice infected with Toxoplasma gondii

目的观察RH株弓形虫感染小鼠血清在体外对小鼠B16黑色素瘤细胞增殖以及凋亡的影响。方法B16细胞在含5%、10%和20%RH株弓形虫感染小鼠血清RPM1640中培养24和48 h,四甲基氮噻唑蓝(MTT)法检测B16细胞增殖抑制率;在10%、20%感染小鼠血清中培养24 h,Annexin V/PI染色,流式细胞检测细胞凋亡,HE染色观察细胞形态改变。结果正常血清能促进B16细胞增殖。5%、10%和20%弓形虫感染小鼠血清与B16细胞共孵育24和48 h,细胞生长抑制率分别为(9.06±0.36)%(、14.77±0.94)%(、23.74±0.5)%和(14.82±0.77)%(、24.56±1.04)%、(39.77±2.82)%,与对照组比较差异有统计学意义(P<0.05);10%、20%弓形虫感染鼠血清组B16细胞24 h凋亡率分别为(26.01±3.27)%和(44.55±2.87)%,显著高于对照组凋亡率(5.01±2.62)(P<0.05)。弓形虫感染鼠血清作用的B16细胞生长密度明显降低,细胞核固缩、浓染。结论RH株弓形虫感染血清能够抑制B16细胞增殖并促进其凋亡。

Objective To observe the proliferation and apoptosis of B16 melanoma cells in the sera of mice infected with RH strain Toxoplasma gondii in vitro. Methods The B16 cells were incubated with RPMI1640 containing 5%,10%,and 20% infected mice sera for 24 h and 48 h in vitro,and the rate of growth inhibition was measured with MTT.The B16 cells were incubated with 10% and 20% infected mice sera for 24 h,the rate of cell apoptosis was examined by flow cytometry with Annexin V/propidium iodide fluorescent staining,and the morphological changes in B16 cells were observed by HE staining. Results The sera of the control promoted B16 cell proliferation.B16 cells were treated with 5%,10%,and 20% infected sera for 24 h and 48 h and had a rate of cell growth inhibition of（9.06±0.36）%,（14.77±0.94）%,（23.74±0.5）%,and（14.82±0.77）%,（24.56±1.04）%,（39.77±2.82）%,respectively;there was a significant difference in comparison to the control（P〈0.05）.The apoptosis rate of B16 cells in 5% and 10% infected sera groups was（26.01±3.27）% and（44.55±2.87）%,respectively;this rate was significantly higher than that in the control group（5.01±2.62,P〈0.05）.The cell density of experimental groups decreased and cells exhibited karyopyknosis and dark staining.Conclusion The sera of mice infected with T.gondii can inhibit the proliferation and promote the apoptosis of B16 cells in vitro.