化疗耐药相关蛋白在舌鳞状细胞癌中的表达和意义

目的：多药耐药相关蛋白1（MRP1）、肺耐药相关蛋白（LRP）、拓扑异构酶Ⅱβ（TOPOUp）和B细胞淋巴瘤2（BCL2）是公认的肿瘤化疗耐药相关蛋白。本研究主要评估这些耐药相关蛋白在舌鳞癌临床标本和体外化疗耐药模型中的表达差异．以及与舌鳞癌临床病理分期的相关性。方法：采用免疫组织化学法检测65例舌鳞癌标本中MRP1、LRP、TOPOIIB和BCL2的表达，分析其与临床病理指标的相关性。采用顺铂逐步诱导SCC15细胞建立体外化疗耐药模型．MTY法检测细胞药敏性改变．免疫荧光检测耐药蛋白的表达变化，蛋白印迹法检测临床标本与舌鳞癌细胞中耐药蛋白的表达差异。应用SPSS16．0软件包对数据进行Mann-WhitnevU检验、Kruskal—WallisH检验、Spearman相关性分析和单因素方差分析。结果：舌鳞癌组织中MRP1、LRP和BCL2高表达，而TOP0118的表达低于癌旁非肿瘤上皮组织（P〈0．05）：MRP1和TOPOIIB的表达与临床分期、淋巴结转移和组织学分级相关，LRP的表达与组织学分级相关（P〈0．05）。SCC15／CDDP耐药细胞株中，MRP1、LRP和BCL2的表达高于舌鳞癌组织，而TOPOIIB的表达显著低于舌鳞癌组织。结论：舌鳞癌标本中MRP1、LRP、BCL2的高表达和TOPOⅡβ的低表达与肿瘤分化程度和化疗耐药相关．提示舌鳞癌患者中存在原发性耐药现象。

The expression and significance of drug resistance-related proteins in tongue squamous cell carcinoma

PURPOSE： Multiple drug resistance protein 1 （MRP1）, lung resistance protein （LRP）, topoisomerase Ⅱβ （TOPO Ⅱβ） and B-cell lymphoma 2 （BCL2） are well-known drug resistance-related proteins in various human malignancies. The aim of this study was to evaluate the expression of these proteins both in clinical tongue squamous cell carcinoma （TSCC） samples and experimental drug-resistance model, and further to determine the relationship between them and the clinico-pathological features of TSCC. METHODS： MRP1, LRP, TOPOⅡβ, and BCL2 expression was examined by immunohistochemistry in specimens from 65 patients with TSCC. A cisplatin-resistant cell line, SCC15/ CDDP was established. A MTF-based method was used to analyze the drug sensitivity. Immunofluorescence was used to detect protein expression in cultured cell lines. Western blot was used to compare the protein expressions in specimens and SCC15/CDDP cells. The results were assessed by Mann-Whitney U test, Kruskal-Wallis H test, Spearman correlation and one-way ANOVA with SPSS16.0 software package. RESULTS： We found higher expression of MRP1, LRP, and BCL2 and lower expression of TOPO Ⅱβ in TSCC compared with adjacent non-neoplastic tongue tissues （P〈0.05）. In addition, MRP1 and TopoⅡβ expression was significantly associated with clinical stage, lymph node metastasis andhistological grade, and LRP was significantly associated with histological grade in the samples （P〈0,05）. Moreover, Western blot showed higher expression of MRP1, LRP, and BCL2 and lower expression of Topo 1113 in SCC15/CDDP cells than in clinical samples. CONCLUSION： The results suggest that the high expression of MRP1, LRP, and BCL2 and low expression of TOPO II [3 in patients with TSCC indicate that intrinsic drug resistance may exist in TSCC, and is associated with tumor differentiation and cisplatin resistance. Supported by National Natural Science Foundation of China （81272949, 81072223, 81172567, 81202136） and Program for New Century Excellent Talents of the Ministry of Education of China （NCET- 11-0535）.