丙戊酸钠联合顺氯氨铂对子宫颈癌的作用机制研究  

The study of mechanism of valproate acid combined with cisplatin on cervical cancer

作　　者：曹振平[1] 冯定庆[2] 李彩荣[2] 张丽娜[1] 刘然[1] 凌斌[1,2] 

机构地区：[1]安徽医科大学附属省立医院妇产科,合肥230001 [2]安徽省分子医学重点实验室,合肥230000

出　　处：《安徽医科大学学报》2011年第10期1001-1006,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家"十一五"科技支撑计划(编号:2008BAI57B00)

摘　　要：目的探讨丙戊酸钠(VPA)联合化疗药物顺铂(DDP)对子宫颈癌作用机制。方法采用3 mmol/L VPA及不同浓度(0.5、1、2.5μg/ml)DDP单独或联合作用于子宫颈癌HeLa细胞,对照组仅加溶媒;VPA 300 mg/(kg.d)-1,DDP 2 mg/kg,每周2次,作用于HeLa细胞接种后成瘤的裸鼠;二甲基四氮唑蓝(MTT)检测细胞增殖抑制;RT-PCR检测细胞及移植瘤中p53,p53上调凋亡调控因子(PUMA)及Bax、Bcl-XL的mRNA表达;Western blot检测细胞及移植瘤中组蛋白H3乙酰化(acetylases histone 3,AC-H3)水平及p53蛋白表达。结果 VPA对宫颈癌HeLa细胞株有明显的生长抑制作用,联合DDP后优于VPA及DDP单药组;荷瘤鼠给药28 d后各组肿瘤体积(mm3):对照组3 155.3±238.7,VPA组1 936.7±207.7,DDP组1 349.9±267.5,VPA+DDP组357.3±212.2,用药组和对照组比较差异有统计学意义(P<0.05)。RT-PCR显示VPA可在mRNA水平上增强p53、PUMA及Bax的表达,减弱Bcl-XL的表达,且联合DDP组效果更明显,与对照组及单药组比较差异有统计学意义(P<0.05)。Western blot显示VPA单独及联合DDP均明显提高细胞及移植瘤中AC-H3及p53和激活型半胱氨酸蛋白酶3(active-caspase-3)的蛋白表达,与对照组比较差异有统计学意义(P<0.05)。结论 VPA联合DDP在体外可杀伤子宫颈癌HeLa细胞,体内可抑制荷瘤裸鼠肿瘤体积增长,而AC-H3及p53和active-caspase-3的表达升高可能为其作用机制。Objective To investigate the effect mechanism of valproate acid combined with cisplatin（DDP） of cervical cancer.Methods HeLa cells were treated with VPA（3 mmol/L） and DDP（0.5,1,2.5 μg/ml） alone or combination.The control group was treated with vehicle only.HeLa cells subcutaneously inoculated into nude mice,then treated with VPA 300 mg/（kg·d）-1,DDP 2 mg/kg,twice a week.MTT assay was used to assess proliferation inhibition.The expression of p53,PUMA,Bax,Bcl-XL,acetylated histone H3（AC-H3） were detected by RT-PCR and Western blot.Results The proliferation of HeLa cells were moderately inhibited by VPA,whereas the growth inhibition of cells co-administrated with VPA and DDP was significantly higher than that treated with each drug alone（P0.05）.VPA and DDP were used to treat xenograft tumors in nude mice for up to 28 days and the tumor volume was measured（mm）3,control：3 155.3±238.7,VPA：1 936.7±207.7,DDP： 1 349.9±267.5,VPA＋DDP： 357.3±212.2.The inhibitory effect of VPA＋DDP on growth of tumor xenografts was significantly higher than that treated with control and each drug alone（P0.05）.RT-PCR showed that VPA could enhance the mRNA expression of p53,PUMA and Bax,and reduce the expression of Bcl-XL in HeLa cells and tumor xenografts.VPA alone or combined with DDP induced histone H3 hyperacetylation,upregulation of p53 and active-caspase-3 in HeLa cells and tumor xenografts,while DDP alone regardless of concentration didn′t alter the expression of these genes.Conclusion VPA alone or in combination with DDP can kill HeLa cells effectively in vitro and inhibitor the growth of tumor xenografts,which maybe attributed to the histone H3 hyperacetylation and up-regulation of p53 and active-caspase-3 after treatment.

关 键 词：丙戊酸 顺铂 肿瘤抑制蛋白质P53 半胱氨酸天冬氨酸蛋白酶3 宫颈肿瘤 

分 类 号：R737.33[医药卫生—肿瘤] R341[医药卫生—临床医学]