重组人PDCD5蛋白对干扰素γ诱导人宫颈癌SiHa细胞凋亡的增敏作用的初步探讨

RhPDCD5 protein can enhance the apoptosis of SiHa ceivical cells induced by IFNγ

目的 初步探讨重组人PDCD5蛋白对干扰素γ诱导人宫颈癌SiHa细胞凋亡的增敏作用。方法人宫颈癌SiHa细胞培养于含 1× 1 0、 1× 1 0 2 、 1× 1 0 3 、 1× 1 0 4IU/ml浓度的IFNγ的无血清培养基中 ,培养 2 0、 2 4、4 8、 72h ,以MTT法指征细胞增殖程度 ,选择适宜的作用浓度及时间。将不同浓度的rhPDCD5蛋白与选定浓度的IFNγ同时加入SiHa细胞中共同培养 ,以AnnexinV -PI标记进行流式细胞仪分析 ,检测细胞凋亡程度。结果 单次给予不同浓度的IFNγ ,培养 2 0h ,与对照组相比 ,1× 1 0 3 组、 1× 1 0 4组的MTT值显著下降 (P <0 0 5 ) ,同时测定细胞凋亡率发现 ,1× 1 0 2 、 1× 1 0 3 、 1× 1 0 4IU/mlIFNγ组SiHa细胞的凋亡率分别为 5 6 7%、 1 2 91 %、 2 0 88% ,随干扰素的浓度上升而增加。 1× 1 0 3 IU/mlIFNγ组加入 1 0、 2 0mg/LrhPDCD5蛋白后细胞凋亡率为 2 4 74 %、2 6 89% ,1× 1 0 4IU/mlIFNγ组加入 1 0、 2 0mg/LrhPDCD5蛋白后细胞凋亡率为 32 6 9%、 32 2 7%。结论干扰素可能通过诱导凋亡而抑制SiHa细胞的生长。rhPDCD5蛋白对干扰素γ诱导人宫颈癌SiHa细胞系凋亡有增敏作用。

Objective To study the effect of rhPDCD5 protein on apoptosis of SiHa cervical cancer cells induced by IFNγ.Methods Human SiHa cervical cancer cells were treated with IFNγ 1×10～1×10 4 IU/ml prepared in serum-free DMEM. The cell proliferation was determined after 20,24,48,72 hours of incubation by MTT assay. Appropriate cell density and incubation time were selected for further study. The cells was treated with different concentrations of rhPDCD5 protein in the presence of IFNγ1×10 3 -1×10 4 IU/ml. Annexin V-labeled FACS assay were applied to study the apoptosis of SiHa cervical cancer cells. Results Incubated with IFNγ 1×10 3～1×10 4 IU/ml for 20 hours suppressed the proliferation of human SiHa cervical cancer cells significantly. The SiHa apoptotic cells were 5 67%,12 91% and 20 88%,increasing with the IFNγ of 1×10 2, 1×10 3 and 1×10 4 IU/ml. The apoptotic rote of SiHa cells that treated with 1×10 3IU/ml of IFNγplus 10 mg/L and 20 mg/L of rhPDCD5 protein were 24 74% and 26 89%, respectively. The apoptotic rate of SiHa cervical cancer cells treated with 1×10 4 IU/ml IFNγplus 10 mg/L and 20mg/L of rhPDCD5 protein were 32 69% and 32 27% respectively. Conclusions IFNγ could inhibits proliferation of SiHa cervical cancer cells by inducing apoptosis. RhPDCD5 protein enhances the apoptotic effect induced by IFNγ in SiHa cells.