宫颈鳞状细胞癌对DDP和DDP+5-FU的体外药敏检测及与耐药蛋白表达的关系研究

In vitro drug sensitivity test for cervical squamous cell carcinoma to DDP and DDP+5-FU and the correlation with drug resistance protein expression

目的通过对宫颈鳞状细胞癌进行顺铂(DDP)和DDP+5-氟尿嘧啶(5-FU)体外药敏检测,比较两种化疗方案的体外有效率。同时检测其耐药蛋白P糖蛋白(P-gp)、谷胱甘肽S-转移酶-π(GST-π)、DNA拓扑异构酶Ⅱ(TopoⅡ)、胸苷酸合成酶(TS)的表达,探讨DDP和DDP+5-FU体外药敏与耐药蛋白表达的关系。方法收集35例宫颈鳞状细胞癌患者的新鲜肿瘤组织,采用三磷酸腺苷生物荧光法(ATP-TCA)对DDP和DDP+5-FU进行体外药敏检测,同时采用EnVision二步法检测宫颈鳞状细胞癌组织中耐药蛋白P-gp、GST-π、TopoⅡ、TS的表达,并分析DDP和DDP+5-FU的体外药敏与耐药蛋白P-gp、GST-π、TopoⅡ、TS表达的关系。结果 35例标本均进行药敏试验,DDP的体外有效率为37.14%,与DDP+5-FU的51.43%比较,差异无统计学意义(P> 0.05)。经多因素分析发现,患者年龄、临床分期、分化程度均不为DDP、DDP+5-FU药物敏感性的影响因素(P> 0.05)。P-gp、GST-π、TopoⅡ、TS蛋白阳性表达率分别为57.14%(20/35)、51.43%(18/35)、71.43%(25/35)、57.14%(20/35)。GST-π蛋白阳性表达是宫颈鳞状细胞癌对DDP耐药的影响因素(P=0.002);TS蛋白阳性表达是宫颈鳞状细胞癌对DDP+5-FU耐药的影响因素(P=0.001)。结论 宫颈鳞状细胞癌ATP-TCA法检测DDP+5-FU与单药DDP相比,体外有效率无显著差异。GST-π、TS蛋白阳性表达可用于临床宫颈癌患者对DDP、5-FU化疗耐药的预测指标。

Objective To determine the in vitro drug sensitivity of cervical squamous cell carcinoma(SCC) to cisplatin(DDP) and cisplatin plus 5-fluorouracil(DDP+5-FU) to compare the in vitro efficacy of the two regimens,and to detect the expression of P-glycoprotein(P-gp),glutathione S-transferase-π(GST-π),topoisomerases Ⅱ(topoll),and thymidylate synthase(TS) expression.Method Fresh tumor tissues were collected from 35 patients with cervical SCC,in which the drug sensitivity to DDP and DDP+5-FU was detected in vitro by triphosadenine bioluminescence assay(ATPTCA),at the same time,EnVision,the two-step detection system,was employed to detect the expression of drug resistance proteins including P-gp,GST-π,topoll,and TS.Furthermore,the relationship between the in vitro sensitivity of DDP and DDP+5-FU and drug resistance proteins P-gp,GST-π,topoll,and TS was assessed.Result ATP-TCA drug sensitivity test was performed on 35 cervical SCC specimens,indicating an in vitro response rate to DDP of 37.14%,which was numerically lower than that of DDP+5-FU at 51.43%,but there was no significant difference(P>0.05).Multivariate analysis revealed that,age,clinical stage,differentiation degree were not factors influencing the drug sensitivity of DDP,DDP+5-FU(P>0.05).The positive expression rate of P-gp,GST-π,topoll,and TS proteins were 57.14%(20/35),51.43%(18/35),71.43%(25/35),and 57.14%(20/35),respectively.Positive expression of GST-π protein was correlated with drug resistance of cervical SCC cells to DDP(P=0.002);meanwhile,TS protein expression was similarly related to drug resistance to DDP+5-FU of cervical SCC cells(P=0.001).Conclusion No significant difference regarding the in vitro efficacy of DDP+5-FU versus DDP alone was demonstrated by ATP-TCA assay.Positive expression of GST-π and TS proteins are potentially predictive for the chemoresistance cervical cancer patients to DDP,5-FU.