Cytopathologic study of intrinsic and acquired drug resistance of human ovarian cancer -the role of YB-1 and cMOAT/MRP2 gene-

目的：第一项研究的目的是通过培养细胞系和手术标本阐明核YB-1与人卵巢癌顺铂耐药之间的关系。方法：采用Western blot方法检测顺铂敏感和耐药卵巢癌细胞株细胞内YB-1的定位。此外，35对来自同一患者的原发性和匹配性复发性卵巢癌的手术标本通过免疫组织化学染色评估其核YB-1表达。结果：核和细胞质提取物的Western blot分析表明，顺铂耐药细胞的核YB-1表达明显高于敏感亲本细胞。免疫组化分析显示，10对病例由原发病灶核YB-1阴性变为复发病灶核YB-1阳性，仅有2对病例由阳性逆转为阴性。结论：YB-1…More的表达可能是顺铂敏感性的有效预测指标和/或顺铂二线化疗治疗复发性卵巢癌的靶分子。目的：第二项研究的目的是通过肿瘤样本中ABC转运蛋白的相对mRNA表达水平来表征具有不良临床结果的患者，并检查每种ABC转运蛋白的相对mRNA水平是否可以作为晚期卵巢癌无进展生存期的有用预测指标。方法：采用实时逆转录- pcr技术检测30例原发性卵巢浆液性乳头状腺癌患者的肿瘤样本中MDR1、MRP1、MRP2和MRP3 mRNA的表达，并评估其与临床预后的相关性。30例患者根据减脂手术及铂基化疗后的临床结果分为3组：8例为不良组；11人被归为有利组；中间组11例。结果：三组间MRP1、MRP3相对mRNA水平差异有统计学意义，经多次比较，不利组MRP1、MRP3 mRNA水平显著高于有利组。MRP1的相对mRNA表达量与MRP3的相对mRNA表达量呈显著相关。在30例浆液性乳头状腺癌患者中，单因素和多因素分析表明，MRP1相对mRNA表达水平高与无进展生存期短显著相关。结论：在晚期卵巢浆液腺癌患者中，这些结果表明，肿瘤样本中MRP1和MRP3 mRNA协同表达水平升高是患者预后不良的特征。此外，较高水平的MRP1 mRNA表达可以作为无进展生存期较短的有用预测指标。少

Purpose: The aim of the first study was to elucidate the association between nuclear YB-1 and cisplatin resistance in human ovarian cancer using cultured cell lines and surgical specimens. Methods: Intracellular YB-1 localization was examined by Western blot analysis for both cisplatin sensitive and resistant human ovarian cancer cell lines. Moreover, 35 pairs of surgical specimens derived from primary and matched recurrent ovarian cancers of the same patient were evaluated for their nuclear YB-1 expression by immunohistochemical staining. Results: Western blot analysis for nuclear and cytoplasmic extracts indicated that cisplatin-resistant cells showed much higher nuclear YB-1 expression than sensitive parental cells. Immunohistochemical analysis showed that 10 paired cases turned from negative nuclear YB-1 in primary lesions to positive nuclear YB-1 in recurrent lesions, whereas only two paired cases showed a reverse turn from positive to negative. Conclusion: The expression of YB-1 … More might be a useful predictive marker indicating cisplatin sensitivity and/or a target molecule to treat recurring ovarian cancers by cisplatin-based second-line chemotherapy.Purpose: The aim of the second study was to characterize patients with unfavorable clinical outcome by the relative mRNA levels of ABC transporter expression in their tumor samples and to examine whether relative mRNA levels of each of the ABC transporter can be a useful predictor of progression-free survival in advanced ovarian carcinoma. Methods: We examined tumor samples taken from 30 patients with primary serous papillary adenocarcinoma of the ovary for the expression of MDR1, MRP1, MRP2, and MRP3 mRNA using real-time reverse transcription-PCR, and we evaluated its correlation with clinical outcome. All 30 patients were divided into three groups according to clinical outcome after debulking surgery and platinum-based chemotherapy: 8 patients were classified into the unfavorable group; 11 were classified into the favorable group; and 11 were classified into intermediate group. Results: The relative mRNA levels of MRP1 and MRP3 were significantly different among the three groups, and the mRNA levels of MRP1 and MRP3 in the unfavorable group were significantly higher than those in the favorable group by multiple comparison. The relative mRNA levels of MRP1 expression were significantly correlated with those of MRP3 expression. In the 30 patients with serous papillary adenocarcinoma, univariate and multivariate analysis demonstrated that the high relative mRNA levels of MRP1 expression were significantly correlated with a short period of progression-free survival. Conclusion: In patients with advanced ovarian serous adenocarcinoma, these results suggest that increased levels of coordinated MRP1 and MRP3 mRNA expression in their tumor samples characterize patients with an unfavorable outcome. Furthermore, a higher level of MRP1 mRNA expression can be a candidate for a useful predictor of a shorter period of progression-free survival. Less