全反式维甲酸联合长春新碱对视网膜母细胞瘤细胞增生的抑制作用  

Inhibitory effect of all trans retinoic acid with vincristine on the proliferation of retinoblastoma cells

作　　者：白海霞[1] 李彬[1] 张浩[2] 顼晓琳[1] 申令[3] 畅颖[1] 

机构地区：[1]首都医科大学附属北京同仁医院北京同仁眼科中心北京市眼科研究所眼科与视觉科学北京市重点实验室,北京100005 [2]首都儿科研究所附属儿童医院眼科,北京100020 [3]哈尔滨医科大学附属第二医院眼科,150001

出　　处：《中华实验眼科杂志》2015年第5期419-423,共5页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81172393）

摘　　要：背景 肿瘤细胞对传统化学治疗药物产生耐药性是视网膜母细胞瘤（RB）化学疗法失败的主要原因,如何提高化学治疗药物的敏感性是RB治疗过程中亟待解决的问题,已有研究表明全反式维甲酸（ATRA）能够抑制肿瘤细胞的生长,但其是否能够提高肿瘤细胞对传统化学治疗药物的敏感性尚不清楚.目的 观察ATRA联合长春新碱对SO-RB50细胞增生的抑制作用. 方法 对SO-RB50细胞进行常规培养,培养液中分别添加不同浓度的ATRA或不同质量浓度的长春新碱,用细胞计数试剂盒-8（CCK-8）法分别测定ATRA及长春新碱作用48 h后对SO-RB50细胞的半数抑制浓度（IC50）.将常规培养的SO-RB50细胞分为正常对照组、ATRA组、长春新碱组和联合用药组,分别将不同药物根据IC50量添加至培养液进行细胞培养,采用CCK-8法于培养后每24小时检测细胞增生情况,连续测定6d,绘制细胞生长曲线.用流式细胞仪检测各组细胞用药72h后不同细胞周期的比例,annexin V/碘化丙啶（PI）法检测各组药物处理48 h后SO-RB50细胞的凋亡情况. 结果 ATRA作用于SO-RB50细胞的IC50值为12.84 μmol/L,长春新碱为0.11 μg/ml.正常对照组SO-RB50细胞生长曲线随着培养时间的延长逐渐上升,但ATRA和长春新碱处理后细胞增生曲线平缓,以联合用药组细胞增生量最低,曲线最平缓.药物作用48 h,正常对照组、ATRA组、长春新碱组和联合用药组细胞的吸光度（A450）分别为1.078±0.022、0.611 ±0.038、0.596±0.031和0.483±0.030,药物作用72 h分别为1.380±0.021、0.799±0.016、0.668±0.041和0.532±0.033,总体比较差异有统计学意义（F分组=1 115.207,P=0.000;F时间=257.781,P=0.000）,ATRA组、长春新碱组和联合用药组A450值均明显低于正常对照组,差异均有统计学意义（均P=0.000）.SO-RB50细胞经药物处理后72 h,各组细胞周期发生变化,与正常对照组比较,ATRA组G0/G1期细胞比例显著增加,S期细胞百�Background Drug resistance is the main cause of failure after chemotherapy of retinoblastoma （RB）,and how to improve the sensitivity of RB cells to chemotherapic drug become an urgent issue.All trans retinoic acid （ATRA） can inhibit the growth of tumor cells.However,whether ATRA increases the sensitivity of RB cells to chemotherapic drug is unclear.Objective This study aimed to investigate the inhibitory effect of ATRA with vincristine on the proliferation of SO-RB50 cells.Methods SO-RB50 cells were cultured routinely.Different concentrations of ATRA or vincristine were added into the medium for 48 hours for the determination of IC50 by cell counting kit-8 （CCK-8） method.Cultured cells were divided into normal control group,ATRA group,vincristine group and combined drug group.The cells were treated by ATRA or vincristine with the dose of IC50,and the proliferation of the cells was detected every day at the 24-hour interval for 6 consecutive days.The percentage of the cells in different cell cycles was analyzed 72 hours after treatment using flow cytometry.Cell apoptosis rate was detected and calculated 48 hours after treatment by annexin V/propidium iodide （PI） method.Results The IC50 of ATRA was approximately 12.84 μ mol/L,and that of vincristine was 0.11 μg/ml.The growth curve of SO-RB50 cells was gradually raised as the lapse of time,but the curves were relatively low in the ATRA group,vincristine group and combined drug group,with the lowest curve in the combined drug group.The proliferation values of the cells （A450）were 1.078±0.022,0.611 ±0.038,0.596 ±0.031 and 0.483 ±0.030 in 48 hours after treatment,and those in 72 hours were 1.380± 0.021,0.799 ± 0.016,0.668 ± 0.041 and 0.532 ± 0.033 in normal control group,ATRA group,vincristine group and combined drug group,showing significant differences among the groups and various time points （Fgroup =1115.207,P =0.000;Ftime =257.781,P =0.000）.The A450 values of the ATRA group,vincristine group and combined drug group were significan

关 键 词：全反式维甲酸 长春新碱 视网膜母细胞瘤 增生 细胞周期 凋亡 

分 类 号：R739.7[医药卫生—肿瘤]