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目的 探讨氟哌啶醇 (Hal)对人红白血病耐药细胞K5 6 2 /Dox的逆转耐药作用 ,以及对P 糖蛋白 (P gp)和肿胀激活的氯离子通道的影响。方法 应用乳酸脱氢酶法 (LDH) ,测定Hal对瘤细胞增殖的抑制作用。以半定量逆转录聚合酶链反应 (RT PCR) ,分析经Hal处理后 3种耐药基因mRNA表达的变化。将瘤细胞荷载氯离子敏感染料MQAE后 ,以荧光分光光度计测定低渗环境中Hal对K5 6 2 /Dox细胞肿胀激活的氯离子通道的影响 ;应用ZM库尔特血球计数仪及 2 5 6频道测定仪 ,测定低渗环境中瘤细胞的体积变化 ,以判断Hal对细胞调节性体积减小 (RVD)的影响。结果 Hal对K5 6 2 /Dox细胞的耐药性具有明显的逆转作用 ,在 12 .5 0 ,6 .2 5和 3.12 μmol/L浓度时 ,其对K5 6 2 /Dox细胞耐药性的逆转倍数分别为 8.6 1,4 .35和 2 .2 5。RT PCR结果显示 ,用 12 .5 0 μmol/LHal处理K5 6 2 /Dox细胞后 ,P gp和多药耐药相关蛋白 (MRP)mRNA表达水平均降低 ,并呈现时间依赖性 ,分别较原水平下降 76 .3%和 6 4 .6 % (P <0 .0 5 ) ;谷胱甘肽硫转移酶π(GSTπ)mRNA的表达水平于用药后第 2天下降6 6 .1% ,第 3天回升 (P <0 .0 5 )。K5 6 2 /Dox细胞的氯离子浓度检测结果显示 ,单纯低渗刺激可使K5 6 2 /Dox细胞中MQAE荧光强度下降 (34.4 6± 5 .91
Objective To investigate the reversal drug resistance of haloperidol (Hal) to human erythroleukemia cell line K5 6 2 / Dox and its effect on P gp and swelling activated chloride channel. Methods Lactate dehydrogenase (LDH) was used to determine the inhibitory effect of Hal on tumor cell proliferation. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to analyze the mRNA expression changes of the three drug-resistance genes after treatment with Hal. Tumor cells were loaded with chloride sensitive dye MQAE, the fluorescence spectrophotometer Hal in K562 / Dox cells in the determination of the impact of swelling on chloride channel K562 / Dox; using ZM Coulter hematocrit and 256 Channel analyzer to determine the volume change of tumor cells in hypotonic environment in order to judge the effect of Hal on cell-mediated reduction of volume (RVD). Results Hal had a significant reversal of drug resistance in K5 6 2 / Dox cells, and reversed the drug resistance of K5 6 2 / Dox cells at 12.5, 6.25 and 3.12 μmol / L Multiples were 8.6 1,4 .35 and 2 .25 respectively. RT-PCR results showed that the mRNA expression levels of P gp and multidrug resistance-related protein (MRP) in K562 / Dox cells treated with 12.5 μmol / L LH were decreased and presented in a time-dependent manner Decreased by 76.3% and 6.46% respectively (P <0.05). The expression of GSTπ mRNA decreased by 61.6% Day rebound (P <0. 05). The chloride ion concentration of K5 6 2 / Dox cells showed that the fluorescence intensity of MQAE decreased in K5 6 2 / Dox cells (34.4 6 ± 5 .91