CAJ | 학술논문

目的探讨细胞膜抗原富含脯氨酸和谷氨酰胺的剪切因子(SFPQ)在多柔比星(ADR)耐药的白血病细胞中的作用.方法以急性髓系白血病细胞株HL-60及其ADR耐药株HL-60/ADR细胞为研究对象,通过免疫荧光法比较HL-60和HL-60/ADR细胞膜SFPQ表达丰度；应用单抗5D12封闭细胞膜SFPQ,通过MTT法测定5D12对HL-60、HL-60/ADR细胞增殖的影响,计算ADR的半数抑制浓度(JC50)和5D12对细胞增殖的促进率；流式细胞术检测5D12对罗丹明在细胞内蓄积的影响.结果 SFPQ在HL-60细胞膜表达高于HL-60/ADR细胞.5D12封闭膜SFPQ作用后,ADR作用HL-60细胞48 h的IC50由(0.19±0.03) μg/ml升高至(0.95 ±0.13) μg/ml,HL-60/ADR细胞的JC50由(4.41±2.42) μg/ml升高至(21.33±4.26) μg/ml.1、3、5、8、12 μg/ml的5D12作用细胞96 h后, HL-60细胞增殖促进率分别为(9.12±2.02)％、(16.63±0.92)％、(19.04±0.25)％、(24.17±0.53)％、(34.04±3.20)％,HL-60/ADR细胞分别为(7.40±2.23)％、(8.72±2.38)％、(10.47±3.78)％、(11.57±1.49)％、(13.97±0.91)％.结论核蛋白SFPQ在HL-60细胞膜表达丰度高于HL-60/ADR细胞；该蛋白在细胞膜易位表达不改变药物在细胞内的蓄积,通过抑制细胞增殖维持HL-60细胞对ADR的敏感性.
목적 탐토세포막항원부함포안산화곡안선알적전절인자(SFPQ)재다유비성(ADR)내약적백혈병세포중적작용.방법 이급성수계백혈병세포주HL-60급기ADR내약주HL-60/ADR세포위연구대상,통과면역형광법비교HL-60화HL-60/ADR세포막SFPQ표체봉도；응용단항5D12봉폐세포막SFPQ,통과MTT법측정5D12대HL-60、HL-60/ADR세포증식적영향,계산ADR적반수억제농도(JC50)화5D12대세포증식적촉진솔；류식세포술검측5D12대라단명재세포내축적적영향.결과 SFPQ재HL-60세포막표체고우HL-60/ADR세포.5D12봉폐막SFPQ작용후,ADR작용HL-60세포48 h적IC50유(0.19±0.03) μg/ml승고지(0.95 ±0.13) μg/ml,HL-60/ADR세포적JC50유(4.41±2.42) μg/ml승고지(21.33±4.26) μg/ml.1、3、5、8、12 μg/ml적5D12작용세포96 h후, HL-60세포증식촉진솔분별위(9.12±2.02)％、(16.63±0.92)％、(19.04±0.25)％、(24.17±0.53)％、(34.04±3.20)％,HL-60/ADR세포분별위(7.40±2.23)％、(8.72±2.38)％、(10.47±3.78)％、(11.57±1.49)％、(13.97±0.91)％.결론 핵단백SFPQ재HL-60세포막표체봉도고우HL-60/ADR세포；해단백재세포막역위표체불개변약물재세포내적축적,통과억제세포증식유지HL-60세포대ADR적민감성.
Objective To investigate the anti-leukemia adriamycin (ADR) effect of membrane protein Splicing factor proline/glutamine-rich (SFPQ).Methods HL-60 and its adriamycin-resistant cells HL-60/ADR were cultured in vitro.Expression of SFPQ on HL-60 and HL-60/ADR cell membranes were examined by immunofluorescence.McAb 5D12 was used to block membrane SFPQ protein activity.ADR susceptibility and cell proliferation were analyzed by MTT assay.IC50 values of ADR in HL-60 and HL-60/ADR cell lines and up-regulations in cell proliferation induced by 5D12 were calculated.Intracellular accumulation of rhodamine in HL-60 and HL-60/ADR cells were measured using fluorescence-activated cell sorting.Results Expression of SFPQ on cell membrane was higher in HL-60 cells compared to the HL-60/ADR cell line.After membraneblocking with 5D12,ADR sensitivity was decreased in vitro compared with the untreated cells [the 48 h IC50 value,HL-60 cell line (0.19±0.03) μg/ml vs (0.95±0.13) μg/ml,HL-60/ADR cell line (14.41±2.42) μg/ml vs (21.33±4.26) μg/ml].Blocking of membrane SFPQ by 5D12 did not affect the intracellular accumulation of rhodamine in these cells,however,5D12 induced HL-60 and HL-60/ADR cell proliferation,following 1,3,5,8 and 12 μg/ml 5D12 treatment for 96 h were (9.12±2.02) ％,(16.63±0.92) ％,(19.04±0.25) ％,(24.17±0.53) ％,(34.04±3.20) ％ (HL-60),and (7.40±2.23) ％,(8.72±2.38) ％,(10.47±3.78) ％,(11.57±1.49) ％,(13.97±0.91) ％ (HL-60/ADR),respectively.Conclusion Nuclear protein-SFPQ contributes to HL-60′ ssensitivity to adriamycin by increasing its surface expression and promoting cell proliferation,but the protein has no significant effect on the intracellular accumulation of the drug.