鞣酸对LLC/cMOAT细胞多药耐药性的逆转机制  

Reversal Mechanism of Tannic Acid on Multidrug Resistant of LLC/cMOAT Cells

作　　者：蒋旭琴[1] 包明红[1] 梅晓冬[1] 

机构地区：[1]安徽医科大学附属省立医院呼吸内科,合肥230001

出　　处：《肿瘤防治研究》2008年第9期630-634,共5页Cancer Research on Prevention and Treatment

摘　　要：目的本研究探讨鞣酸(Tannic acid,TA)对LLC/cMOAT细胞多药耐药性的逆转作用及其可能机制。方法采用MTT法检测LLC/CMV和LLC/cMOAT细胞对各种化疗药物的敏感性以及在不同浓度的TA处理下细胞的存活状态;确定TA的非细胞毒性浓度以及在该浓度下各梯度浓度处理时细胞的存活率;采用流式细胞仪技术检测使用非细胞毒性浓度的TA处理前后LLC/cMOAT细胞内柔红霉素(DNR)浓度变化、细胞周期阻滞及对凋亡的影响。结果LLC/cMOAT细胞对羟基喜树碱(CPT-11),阿霉素(ADM),顺铂(DDP)和长春新碱(VCR)均有不同程度的耐药,其耐药倍数分别是2.08、3.02、4.01和9.31,对依托泊苷(VP-16)、紫衫醇(TAX)无明显耐药,10.0μmol/L浓度以下的TA对LLC/CMV和LLC/cMOAT细胞无明显细胞毒性,2.5、5.0、10.0μmol/L浓度的TA能显著降低LLC/cMOAT细胞的IC50(P<0.05),其逆转倍数分别为5.09、6.33、7.76倍;细胞内DNR荧光强度随着TA浓度的增高而明显增强;TA作用细胞12h可使G1期细胞数百分比由(46.35±3.74)%增至(66.43±5.87)%,阻滞细胞于G1期;TA与VCR联合处理细胞24h和48h时凋亡率分别为12.1%和19.0%,与处理前(1.65%)相比提高明显;使用2.5μmol/L、5.0μmol/L和10.0μmol/L浓度的TA与VCR共同处理细胞24h后,凋亡率由处理前的4.96%分别提高到11.2%、17.9%和25.1%。结论TA能部分逆转LLC/cMOAT细胞的多药耐药,显著提高耐药细胞内DNR荧光强度并呈浓度依赖性,阻滞细胞周期于G1期,并可诱导细胞凋亡,呈时间、剂量依赖性,其机制可能与抑制化疗药物外排、增加细胞内药物浓度以及诱导细胞凋亡有关。Objective To investigate the reversal effects of tannic acid（TA） on the multidrug resistance of LLC/cMOAT cells and the possible mechanism. Methods MTT assay was used to determine the sensitivity of LLC/CMV and LLC/cMOAT cells to the various kinds of chemotherapeutics, and the activity of LLC/CMV and LLC/cMOAT cells treated with different concentrations of TA. The non-cytotoxic concentrations of TA were determined for the ceils and the reversal effects of the chemotherapeutics were observed. The intracellular concentration of DNR, the cell cycle distribution and the apoptosis rate of the cells treated with different concentrations of TA were determined by flow cytometry（FCM）. Results LLC/cMOAT cells were resistant to CPT-11, ADM, DDP, VCR to a certain extent, and not resistant to VP-16,TAX. At 10. 0μmot/L and below, TA was not significantly cytotoxic to LLC/CMV and LLC/ cMOAT cells. TA at the concentration of 2. 5,5.0,10. 0 μmol/L may remarkably decrease IC50 of LLC/ cMOAT cells（P（0. 05）. The intracellular DNR fluorescence intensity in LLC/cMOAT cells was significantly enhanced with the increase of TA concentration. The cells at G, stage increased from （46. 35 ± 3.74）% to （66. 43 ±5.87） % when treated with TA at 5.0 μmol/L for 12 hours. The cell apoptosis was enhanced in a time-and concentration-dependent manner by treating with TA. Conclusion TA is able to reverse the multidrug resistance of LLC/cMOAT cells and may remarkably raise drug concentrations in concentration-dependent manner,TA is able to block cell cycle at G1 stage, and its function of inducing apoptosis acts in time-and close-depended manner. The mechanism may involve the decrease of chemotherapeutics excretion,the increase of intracellular drug concentration, growth arrest at G1 and apoptosis.

关 键 词：鞣酸 cMOAT 多药耐药 逆转 细胞凋亡 

分 类 号：R730.53[医药卫生—肿瘤]