ERCC1 mRNA表达与恶性胸腹腔积液肿瘤细胞对顺铂体外药敏相关性的研究

目的：分析切除修复交叉互补基因l（excision repair egoS8 complementafion group 1,ERCC1）在恶性胸腹腔积液肿瘤细胞中的表达,探讨ERCC1 mRNA表达与患者顺铂耐药的相关性.方法：收集26例经病理确诊的恶性胸腹腔积液,分离原代肿瘤细胞,CCK8体外药物敏感性试验检测顺铂对原代肿瘤细胞的抗肿瘤作用,荧光定量RT-PCR检测细胞中药物疗效相关基因ERCC1的相对表达水平.结果：ERCCI mRNA在恶性胸腹腔积液原代肿瘤细胞中的表达水平与临床特征无相关性,但与顺铂敏感件呈负相关,低表达者对顺铂敏感性高（P=0.01）.结论：ERCC1 mRNA表达水平可以作为预测转移性胸腹腔积液患者铂类药物化疗疗效的指标之一.     

ERCC1 mRNA表达与恶性胸腹腔积液肿瘤细胞对顺铂体外药敏相关性的研究

目的:分析切除修复交叉互补基因l(excision repair egoS8 complementafion group 1,ERCC1)在恶性胸腹腔积液肿瘤细胞中的表达,探讨ERCC1 mRNA表达与患者顺铂耐药的相关性.方法:收集26例经病理确诊的恶性胸腹腔积液,分离原代肿瘤细胞,CCK8体外药物敏感性试验检测顺铂对原代肿瘤细胞的抗肿瘤作用,荧光定量RT-PCR检测细胞中药物疗效相关基因ERCC1的相对表达水平.结果:ERCCI mRNA在恶性胸腹腔积液原代肿瘤细胞中的表达水平与临床特征无相关性,但与顺铂敏感件呈负相关,低表达者对顺铂敏感性高(P=0.01).结论:ERCC1 mRNA表达水平可以作为预测转移性胸腹腔积液患者铂类药物化疗疗效的指标之一.     

恶性胸腔/腹腔积液RRM1 mRNA表达水平和吉西他滨体外药敏的关系

目的:分析恶性胸腔/腹腔积液中肿瘤细胞核糖核苷酸还原酶M1亚基(ribonucleotide reductase M1,RRM1)基因的表达水平与吉西他滨体外药物敏感性的关系。方法:收集24例经病理确诊的Ⅳ期恶性肿瘤患者的恶性胸腔/腹腔积液,分离原代肿瘤细胞,采用CCK8(cell counting kit8)法体外药物敏感试验检测原代肿瘤细胞对吉西他滨药物敏感性;实时荧光定量PCR检测原代肿瘤细胞RRM1 mRNA的相对表达水平。结果:在恶性胸腔/腹腔积液中,肿瘤细胞的RRM1 mRNA的相对表达水平与恶性肿瘤患者的临床特征无明显相关性,但是与吉西他滨药物敏感性呈负相关(r=-0.497,P=0.013);既往曾使用化疗药物的恶性肿瘤患者恶性胸腔/腹腔积液原代肿瘤细胞对吉西他滨敏感性低于未化疗患者(42.94%vs67.88%,P=0.006)。结论:恶性胸腔/腹腔积液中肿瘤细胞RRM1 mRNA的相对表达水平与吉西他滨体外药物敏感性相关。     

恶性胸腔/腹腔积液RRM1 mRNA表达水平和吉西他滨体外药敏的关系

目的：分析恶性胸腔/腹腔积液中肿瘤细胞核糖核苷酸还原酶M1亚基（ribonucleotide reductase M1,RRM1）基因的表达水平与吉西他滨体外药物敏感性的关系。方法：收集24例经病理确诊的Ⅳ期恶性肿瘤患者的恶性胸腔/腹腔积液,分离原代肿瘤细胞,采用CCK8（cell counting kit8）法体外药物敏感试验检测原代肿瘤细胞对吉西他滨药物敏感性;实时荧光定量PCR检测原代肿瘤细胞RRM1 mRNA的相对表达水平。结果：在恶性胸腔/腹腔积液中,肿瘤细胞的RRM1 mRNA的相对表达水平与恶性肿瘤患者的临床特征无明显相关性,但是与吉西他滨药物敏感性呈负相关（r=-0.497,P=0.013）;既往曾使用化疗药物的恶性肿瘤患者恶性胸腔/腹腔积液原代肿瘤细胞对吉西他滨敏感性低于未化疗患者（42.94%vs67.88%,P=0.006）。结论：恶性胸腔/腹腔积液中肿瘤细胞RRM1 mRNA的相对表达水平与吉西他滨体外药物敏感性相关。     

恶性胸腹水中肿瘤细胞BRCA1和β-tubulin Ⅲ的表达与多西紫杉醇药物敏感性的关系

目的:疗效相关分子生物标记的探索是实现个体化治疗的关键所在。由于肿瘤原发灶与转移灶肿瘤细胞在分子水平的显著差异及转移性恶性肿瘤的难治性,本研究旨在探索肿瘤特殊转移灶即恶性胸/腹水中肿瘤细胞的BRCA1和β-tubulinⅢmRNA表达水平与多西紫杉醇(TXT)药物敏感性的关系。方法:前瞻性地收集41例进展期肿瘤的恶性胸水/腹水,分离肿瘤细胞后ATP-TCA方法检测TXT敏感性;实时荧光定量RT-PCR检测的BRCA1和β-tubulinⅢmRNA表达。结果:在非小细胞肺癌(NSCLC)组(P=0.008)和胃癌组(P=0.032)的恶性胸/腹腔积液中肿瘤细胞的BRCA1 mRNA表达与TXT体外药物敏感正相关。β-tubulinⅢmRNA表达仅在NSCLC患者的恶性胸腔积液中肿瘤细胞与TXT药物敏感负相关(P=0.022)。联合检测BRCA1和β-tubulinⅢ对于TXT药物敏感性的预测没有交互作用(P>0.05)。结论:在NSCLC和/或胃癌患者的恶性胸/腹腔积液中,BRCA1和β-tubulinⅢ表达与TXT敏感性相关且前者可能比后者更有意义。对于伴有恶性积液的进展期NSCLC和/或胃癌患者,恶性积液有可能成为一合适的标本进行分子生物标记的检测。     

转移性恶性胸腹腔积液患者肿瘤细胞中乳腺癌易感基因1mRNA的表达与铂类敏感性的关系

目的 研究乳腺癌易感基因1(BRCA1)在转移性恶性胸腹腔积液肿瘤细胞中的表达,并探讨BRCA1基因表达在铂类化疗疗效中的预测作用.方法 收集31例经病理确诊的恶性胸腹腔积液标本,分离原代肿瘤细胞.采用细胞计数试剂盒(CCK8)体外药物敏感性试验,检测顺铂(DDP)对原代肿瘤细胞的抗肿瘤作用,采用实时荧光定量PCR法检测肿瘤细胞中BRCA1 mRNA的相对表达水平.结果 BRCA1 mRNA在原代肿瘤细胞中的表达水平为0.618(0.014～18.063),DDP对原代肿瘤细胞的半数抑制浓度(IC50)为2.809 μg/ml (0.118～19.439 μg/ml).BRCA1 mRNA的表达及DDP对原代肿瘤细胞的IC50值与患者的年龄、性别、原发肿瘤类型、是否接受化疗以及积液类型均无关(均P＞0.05).BRCA1 mRNA的表达水平与DDP抵抗正相关,BRCA1 mRNA低表达者对DDP的敏感性高(r=0.786,P＜0.001).结论 检测BRCA1 mRNA的表达水平可以为转移性胸腹腔积液患者应用铂类药物化疗提供理论依据.

Abstract：

Objective To explore the mRNA expression of breast cancer susceptibility gene 1 (BRCA1) in tumor cells isolated from malignant pleural and peritoneal effusions, and the predictive role of BRCA1 related to the efficacy of cisplatin-based chemotherapy. Methods Tumor cells were isolated from malignant pleural and peritoneal effusions of 31 cancer patients. The response of these tumor cells to cisplatin was determined by CCK8 assay. Real time quantitative RT-PCR was used to examine the BRCA1 mRNA level in the primary culture cancer cells. Results The expression level of BRCA1 mRNA was 0.618(0.014-18.063)in primary culture tumor cells. The IC50 of DDP was 2.809 μg/ml in the primary culture tumor cells (0.118-19.439 μg/ml). Both BRCA1 mRNA expression and the tumor cells IC50 of DDP were not significantly related with patient age, gender, the type of primary tumor, whether to accept the chemotherapy and effusion type (P>0.05). The level of BRCA1 mRNA was negatively correlated with the chemosensitivity in terms of IC50 of cisplatin (P<0.001). Conclusion Assessment of expression level of BRCA1 mRNA may be useful in predicting the efficacy of cisplatin-based chemotherapy in patients with metastatic malignant effusions.     

恶性胸腹水中肿瘤细胞BRCA1和β-tubulin Ⅲ的表达与多西紫杉醇药物敏感性的关系

目的：疗效相关分子生物标记的探索是实现个体化治疗的关键所在。由于肿瘤原发灶与转移灶肿瘤细胞在分子水平的显著差异及转移性恶性肿瘤的难治性,本研究旨在探索肿瘤特殊转移灶即恶性胸/腹水中肿瘤细胞的BRCA1和β-tubulinⅢmRNA表达水平与多西紫杉醇（TXT）药物敏感性的关系。方法：前瞻性地收集41例进展期肿瘤的恶性胸水/腹水,分离肿瘤细胞后ATP-TCA方法检测TXT敏感性;实时荧光定量RT-PCR检测的BRCA1和β-tubulinⅢmRNA表达。结果：在非小细胞肺癌（NSCLC）组（P=0.008）和胃癌组（P=0.032）的恶性胸/腹腔积液中肿瘤细胞的BRCA1 mRNA表达与TXT体外药物敏感正相关。β-tubulinⅢmRNA表达仅在NSCLC患者的恶性胸腔积液中肿瘤细胞与TXT药物敏感负相关（P=0.022）。联合检测BRCA1和β-tubulinⅢ对于TXT药物敏感性的预测没有交互作用（P〉0.05）。结论：在NSCLC和/或胃癌患者的恶性胸/腹腔积液中,BRCA1和β-tubulinⅢ表达与TXT敏感性相关且前者可能比后者更有意义。对于伴有恶性积液的进展期NSCLC和/或胃癌患者,恶性积液有可能成为一合适的标本进行分子生物标记的检测。     

晚期非小细胞肺癌组织ERCC1和BRCA1表达与铂类化疗敏感性临床研究

目的 吉西他滨联合顺铂是治疗晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的一线化疗方案,但其有效率仅20％～40％,其较低的化疗有效率与肿瘤耐药相关.研究NSCLC组织中切除修复交叉互补基因1(exeision repair cross-compietionl1,ERCC1)和乳腺癌易感基因1(breast cancer susceptibility gene-1,BRCA1)mRNA的表达及其与铂类化疗敏感性的关系.方法 应用实时荧光定量PCR技术检测2009-03-01-2011-04-16武汉钢铁(集团)公司第二职工医院收集的98例晚期NSCLC患者中ERCC1和BRCA1 mRNA的表达情况,分析其与含铂联合治疗方案化疗耐药性的关系.结果 检测98例患者中ERCC1和BRCA1 mRNA的相对表达量,其中位数分别为0.048和0.011,四分位间距IQR分别为0.0058和0.001.ERCC1 mRNA的高表达率为56.1％(55/98),其表达在不同性别、年龄、组织学类型及临床分期中差异无统计学意义,P＞0.05;BRCA1 mRNA的高表达率为64.3％(63/98),其表达在不同性别、年龄、组织学类型及临床分期中差异无统计学意义,P＞0.05.在ERCC1 mRNA低表达组GP方案化疗客观缓解率为48.8％(21/43),而在高表达组化疗客观缓解率为29.1％(16/55),两组之间比较差异有统计学意义,x2 =4.0,P=0.045;在BRCA1 mRNA低表达组GP化疗客观缓解率为57.1％(20/35),在高表达组GP化疗客观缓解率为28.6％(18/63),差异有统计学意义,x2 =7.74,P=0.005;ERCC1和BRCA1 rnRNA联合低表达组中GP化疗客观缓解率为70.4％(19/27),而其联合高表达组GP化疗客观缓解率仅为21.1％(8/38).结论 ERCC1、BRCA1 mRNA高表达患者在使用含铂化疗方案时疗效差,提示ERCC1和BRCA1 mRNA高表达NSCLC患者对铂类具有耐药性.ERCC1和BRCA1mRNA联合低表达患者在使用含铂方案化疗时有效率更高,提示应用ERCC1和BRCA1 mRNA联合检测时,患者可获得更大的益处.     

恶性胸腔积液中ERCC1基因多态性与顺铂化疗敏感性的关系

目的:探讨恶性胸腔积液中ERCC1的基因多态性与顺铂化疗敏感性的相关性。方法:收集70例经病理证实的非小细胞肺癌伴恶性胸腔积液患者,检测胸腔积液中ERCC1基因分型,并给予胸腔灌注顺铂及全身化疗,分析其对顺铂为基础的化疗疗效的影响。结果:ERCC1 T354c基因型频率分别为CC 25.7%、CT 57.1%、TT 17.1%,T354c的TT基因型与CT/CC基因型对顺铂化疗效果的差异有统计学意义(25.0% vs 65.5%,P=0.021）。结论:恶性胸腔积液中ERCC1的基因多态性可能与顺铂化疗敏感性相关。     

RNA干扰ERCC1基因表达对肺腺癌细胞A549/DDP顺铂耐药的影响

背景与目的核苷酸切除修复机制可修复顺铂(DDP)造成的DNA损伤,作为其中的关键酶之一,切除修复交叉互补基因1(excision repair cross-complementing gene 1,ERCC1)的表达可能与DDP耐药有关.本研究旨在探讨小干扰RNA沉默ERCC1基因表达对肺腺癌耐药细胞A549/DDP药物敏感性的影响.方法将体外设计合成针对ERCC1的小分子RNA(siRNA)转染A549/DDP细胞,RT-PCR检测ERCC1 mRNA水平;Western blot检测ERCC1蛋白表达的变化;MTT检测RNA干扰后细胞药物敏感性改变.结果 ERCC1 RNAi干扰后ERCC1 mRNA表达指数降低,明显低于对照组,干扰后ERCC1 mRNA表达抑制率为90.4%.转染ERCC1 siRNA降低TERCC1蛋白的表达水平.MTT显示分别用2μg/mL、4μg/mL、8μg/mL、16μg/mL、32μg/mL顺铂处理细胞48 h,转染ERCC1 siRNA细胞组较对照组敏感性明显增高,干扰前A549/DDP细胞IC50为12.49μg/mL,干扰后A549/DDP细胞IC50下降为9.27 μg/mL.结论 ERCC1siRNA干扰可以降低ERCC1的表达水平,并可提高A549/DDP细胞对顺铂的敏感性.     

ERCC1 and BRCA1 mRNA expression levels in metastatic malignant effusions is associated with chemosensitivity to cisplatin and/or docetaxel

Background  

One of the major challenges in currently chemotherapeutic theme is lacking effective biomarkers for drug response and sensitivity. Our current study focus on two promising biomarkers, ERCC1 (excision repair cross-complementing group 1) and BRCA1 (breast cancer susceptibility gene 1). To investigate their potential role in serving as biomarkers for drug sensitivity in cancer patients with metastases, we statistically measure the mRNA expression level of ERCC1 and BRCA1 in tumor cells isolated from malignant effusions and correlate them with cisplatin and/or docetaxel chemosensitivity.     

ERCC1、BAG-1 及 BRCA1 基因在非小细胞肺癌中的表达及其与预后的关系

目的 探讨ERCC1、BAG-1和BRCA1基因mRNA在接受以铂类为基础辅助化疗的非小细胞肺癌（non-small cell lung cancer,NSCLC）患者中的表达情况及其与预后的关系。方法 85例NSCLC患者中74例给予以铂类为基础的术后辅助化疗,采集全组患者的肿瘤组织和其中34例患者的癌旁组织,应用半定量RT-PCR法,检测其ERCC1、BAG-1和BRCA1基因mRNA的表达情况,回顾性分析患者的临床病理资料,探讨3个基因的表达与患者总生存期（OS）之间的关系。结果 ①ERCC1、BAG-1阴性表达的患者OS显著长于阳性表达的患者（P均=0.001）。ERCC1、BAG-1阴性表达的患者接受铂类化疗的疗效显著优于阳性表达的患者（P=0.002、P=0.001）。②BRCA1阴性表达的患者OS长于阳性表达的患者,但差异无统计学意义（P=0.057）。③多因素分析显示ERCC1、BAG-1可作为NSCLC患者接受铂类化疗的预后指标（P=0.027、P=0.022）。结论 检测ERCC1和BAG-1的表达可作为指导NSCLC患者术后接受铂类化疗及预后评估的指标。     

Combined evaluation of Rad51 and ERCC1 expressions for sensitivity to platinum agents in non-small cell lung cancer

DNA repair enzyme expression in tumor cells possibly affects sensitivity to anti-cancer agents. The aim of this study was to determine the relationship between expression status of DNA repair enzymes and chemosensitivity in patients with non-small cell lung cancer (NSCLC). NSCLC tissues prepared from the surgical specimens of 41 patients were subjected to immunohistochemical analysis for Rad51 and ERCC1 proteins and to a chemosensitivity test using the MTT assay. The relationships between the expression status of the DNA repair enzymes and ex vivo chemosensitivity to various agents were evaluated. A positive expression for Rad51 and ERCC1 was observed in 17 cases (41%) and 20 cases (49%), respectively. The positivity of Rad51 was closely related to a certain histologic type of squamous cell carcinoma and poor differentiation, and the positivity of ERCC1 tended to be related to squamous cell carcinoma. In chemosensitivity tests, sensitivities to CDDP and CBDCA were significantly lower when both 2 enzymes were positive (p = 0.012 and 0.04 in CDDP, 0.014 and 0.03 in CBDCA). Both Rad51 and ERCC1 expressions showed no significant relationship with sensitivities to paclitaxel, etoposide, vinorelbine, gemcitabine, 5-FU, or irinotecan. In conclusion, combined expression of Rad51 and ERCC1 expression is associated with resistance to platinum agents in the ex vivo study of clinical NSCLC, and evaluation of expression status of both DNA repair enzymes would be a predictor for clinical response to platinum-based chemotherapies.