Nakajima Yasuaki, Miyake Satoshi, Tanaka Koji, Ogiya Kazuo, Toukairin Yutaka, Kawada Kenrou, Nishikage Tetsurou, Nagai Kagami, Kawano Tatsuyuki
Department of Esophago-Gastric Surgery, Tokyo Medical and Dental University, Bunkyou-ku, Tokyo 113-8519, Japan.
Cancer Sci. 2004 May;95(5):464-8. doi: 10.1111/j.1349-7006.2004.tb03233.x.
Previously, we demonstrated that CPT-11 is an effective agent against esophageal squamous cell cancers (ESCC), and that the protein level of DNA topoisomerase I can be a predictor for sensitivity to CPT-11 (Jpn J Cancer Res 2001; 92: 1335-41). Here, we describe our search for additional predictors of sensitivity to CPT-11, mainly among cell cycle-regulating proteins, because the cytotoxicity of CPT-11 is significantly correlated with the percentage of ESCC cells in S-phase. To this end, we selected and examined the expressions of 5 proteins involved in G1-S transition, i.e., p53, cyclin D1, p21, p27, and pRB, in 14 ESCC cell lines by western blot analysis. Among these proteins, the expression levels of p21 and pRB showed significant differences that were associated with the IC50 values for CPT-11 (P = 0.0339 and P = 0.0109, respectively). Namely, the expression of p21 or pRB independently could be a good indicator of CPT-11 efficacy in ESCC. In addition, the cell proliferation activities examined by enzyme-linked immunosorbent assay (ELISA) using 5-bromo-2'-deoxyuridine (BrdU) showed a significant correlation with the percentage of total S-phase cells (correlation coefficient = 0.568, P = 0.0324), and an inverse correlation with the IC50 values for CPT-11 (correlation coefficient =-0.601, P = 0.0213). Because, as in the case of DNA topoisomerase I, the cell proliferation activity determined using BrdU shows a close relationship with the MIB-1 labeling index, immunohistochemical studies of p21, pRB, and MIB-1 in resected ESCC specimens and/or biopsy samples could make it possible to predict more precisely the sensitivity of ESCC patients to CPT-11 prior to treatment.
此前,我们证明了CPT - 11是一种治疗食管鳞状细胞癌(ESCC)的有效药物,并且DNA拓扑异构酶I的蛋白水平可以作为预测对CPT - 11敏感性的指标(《日本癌症研究杂志》2001年;92:1335 - 41)。在此,我们主要在细胞周期调节蛋白中寻找对CPT - 11敏感性的其他预测指标,因为CPT - 11的细胞毒性与处于S期的ESCC细胞百分比显著相关。为此,我们通过蛋白质印迹分析选择并检测了14种ESCC细胞系中5种参与G1 - S期转换的蛋白的表达,即p53、细胞周期蛋白D1、p21、p27和pRB。在这些蛋白中,p21和pRB的表达水平显示出与CPT - 11的IC50值相关的显著差异(分别为P = 0.0339和P = 0.0109)。也就是说,p21或pRB的表达各自都可以作为ESCC中CPT - 11疗效的良好指标。此外,使用5 - 溴 - 2'-脱氧尿苷(BrdU)通过酶联免疫吸附测定(ELISA)检测的细胞增殖活性与总S期细胞百分比显著相关(相关系数 = 0.568,P = 0.0324),并且与CPT - 11的IC50值呈负相关(相关系数 = - 0.601,P = 0.0213)。因为,如同DNA拓扑异构酶I的情况一样,使用BrdU测定的细胞增殖活性与MIB - 1标记指数密切相关,对切除的ESCC标本和/或活检样本中的p21、pRB和MIB - 1进行免疫组织化学研究,有可能在治疗前更精确地预测ESCC患者对CPT - 11的敏感性。