氧分压变化对NK92细胞生存及黏附性的影响

The effect of oxygen partial pressure on cell viability and adhesion in vitro

目的探讨体外培养环境中氧分压变化对NK92细胞迁移特性及相关基因表达的影响。方法将NK92细胞株分别置于1%、5%、10%、15%及21%氧,5%CO2环境下培养6小时;采用流式细胞技术检测细胞活力,甲苯胺蓝法检测黏附能力;RT-PCR半定量技术检测迁移相关基因的表达。结果不同氧分压条件下NK92细胞活率无显著区别;1%、5%、10%氧压下NK92细胞黏附促进率显著高于15%及21%氧压组(P<0.05);10%氧压下NK92细胞CCR1、5、6、7及CXCR4 mRNA的水平显著高于常氧(21%O2)对照组(P<0.05);低氧(1%O2)显著下调NK92细胞CCR5、7、CX3CR1及CXCR4基因的表达,上调HIF-1α及CCR1基因的表达。结论体外培养环境中氧分压变化能影响NK92细胞黏附性及相关基因表达的变化,但不影响其生存;NK92细胞低氧环境下生存可能与HIF-1α及CCR1基因表达上调有关。体外细胞功能及药物效应的研究不仅要考虑模拟体内的CO浓度,还应考虑氧环境因素对细胞的影响。

Objective：To test the effect of PO2 on cell viability, adhesive ability and resultant gene expression in NK92 cell. Methods： NK92 cells were cultured under different PO2 （1% ,5 % , 10%, 15% and 21% ） and 5% CO2 for 6 h. Cell viability was analyzed by FACS. Toluidine blue was used to detect cell adhesive ability and RT-PCR was performed to test the gene expression of CCRs, CXCs and HIF-1 ot at mRNA level. Results：There was no significant difference in cell viability cultured under different oxygen tension. Cell adhesive ability was increased when cells were cultured under 1% ,5%, 10% oxygen tension compared with the cells that cultured under 15 % and 21% oxygen tension （ P 〈 0.05 ）. Ten percent oxygen could significantly increase the expression of CCR1,5,6,7 and CXCR4 at mRNA level than the control （ 21% O2 ） （ P 〈 0. 05 ）. Although hypoxia （ 1% O2 ） could inhibit the expression of CCRS, 7 and CXCR4, the expression of HIF-1α and CCR1 was increased under hypoxia circumstance（ P 〈 0. 05 ）. Conclusion ：The variety of oxygen tension in the in vitro culture circumstance could affect the cell adhesive ability and some functional-related gene expression of NK92 cell, but had no significant influence in cell viability. Cell viability may be maintained by the increased expression of HIF-1α and CCR1. The results of our study show that in vitro experiments for cell function research or for medicine efficiency test, it is important to consider the effect of oxygen tension as well as the concentration of CO2.