氟对人成骨样细胞株凋亡的影响

王军舰; 王海彬

doi:10.11847/zgggws2005-21-12-24

氟对人成骨样细胞株凋亡的影响

Effect of NaF on apoptosis in human osteoblast-like cell line

摘要

摘要:
目的研究氟化钠(NaF)对人成骨样细胞株凋亡的影响及其机制。
方法四甲基偶氮噻唑蓝(MTT)法测定细胞的增殖能力, 萤光染剂(Hoechst 33342)染色, DNA梯形(DNA Ladder)琼脂糖电泳, 流式细胞仪分析检测细胞凋亡, RT-PCR半定量分析抗凋亡蛋白Bcl-2和促凋亡蛋白Bax mRNA表达, 显色法检测凋亡相关激酶半胱氨酸蛋白酶3(Caspase-3)的活性。
结果 NaF(4, 6, 8×10^-3mol/L)可以剂量依赖性地抑制细胞增殖(P < 0.01), Hoechst33342染色、琼脂糖电泳和流式细胞仪均可检测到氟对成骨细胞凋亡的诱导作用。RT-PCR结果显示:NaF可以剂量依赖性地抑制Bcl-2 mRNA表达(P < 0.01), 提高Bax mRNA表达(P < 0.01), 并显著增强Caspase-3的活性(P < 0.01)。
结论 NaF可以显著的抑制人成骨样细胞株增殖和诱导其凋亡, 并可能是通过抑制Bcl-2表达, 促进Bax表达和提高Caspase-3活性来诱导人成骨样细胞株凋亡。

Abstract:
Objective To study the apoptotic effect of sodium fluoride(NaF)and its mechanisms in human osteoblast cell line OS-732.
Methods The effects of NaFon osteoblast cell proliferation and apoptosis were detected by methyl thia zolyl tetrazolium(MTT), Hoechst33342 fluorescence staining, agarose gel electrophoresis, flow cytometry(FCM), semi-quantitative RT-PCR and caspase-3 activity assays.
Results After cells were treated with NaF(4, 6, 8×10^-3 mol/L)for 24 h, the proliferation of OS-732 cells measured by MTT methods was inhibited in a dose dependent manner(P < 0.01).The morphological changes and the apoptotic peaks were observed by fluorescence staining and FCM respectively.In addition, the expression of Bcl-2 mRNA decreased whereas the expression of Bax mRNA increased measured by semi-quantitative RT-PCR.The activity of Caspase-3 also increased after cells were treated with NaF(6×10^-3 mol/L)for 24h(P < 0.01).Finally, DNA fragmentation was observed when cells were treated with NaF(4, 6, 8×10^-3 mol/L)for 48 h.
Conclusion NaF inhibited the proliferation of osteoblast cell line OS-732 and induced the cells to undergo apoptosis by increasing the activity of caspase-3, and the expression of Bax while decreasing the expression of Bcl-2.

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