晚期大肠癌ERCC5基因启动子-763A〉G多态性与铂类药物疗效相关性的研究

目的：探讨ERCC5基因启动子区单核苷酸多态性与中国汉族晚期大肠癌奥沙利铂疗效的相关性。方法：收集以奥沙利铂为主化疗的105例晚期大肠癌患者化疗开始前静脉血,提取基因组DNA,应用PCR-LDR方法检测ERCC5基因3个SNP位点-1415C〉T（rs2094258）-7、63A〉G（rs2016073）及-413C〉T（rs943245）多态性,分析不同基因型与疾病控制率、无进展生存期（PFS）的相关性。结果：携带ERCC5-763GG-、763AG和-763AA基因型的患者化疗疾病控制率分别为86.7%、69.2%和52.6%,其中携带-763GG基因型的患者疾病控制率显著高于携带-763AA基因型的患者,P=0.028。携带-763AA基因型的患者中位PFS（36/95例,6个月）低于携带-763AG基因型（48/95例,8个月）及携带-763GG基因型（11/95例,10个月）的患者,差异有统计学意义,χ^2=9.205,P=0.002。-1415C〉T多态性与化疗疾病控制率和PFS之间均无显著相关性。-413C〉T位点未发现遗传多态。结论：ERCC5启动子区-763A〉G单核苷酸多态性与晚期大肠癌奥沙利铂临床疗效相关。     

ERCC5基因+25A＞G多态性与晚期结直肠癌铂类药物疗效的相关性

目的 探讨切除修复交叉互补基因5(ERCC5)基因单核苷酸多态性与中国汉族晚期结直肠癌患者铂类药物疗效的相关性.方法 应用PCR-LDR法检测以奥沙利铂为主化疗的105例晚期结直肠癌患者ERCC5基因位点+25A＞G (rs751402)、+202C＞T (rs2296147)及+372C＞T (rs2296148)的...     

晚期大肠癌患者外周血ERCC1基因多态性与奥沙利铂化疗疗效相关性的研究

目的：研究ERCC1 Asn118Asn（C→T，rs11615）多态性与中国汉族晚期大肠癌患者对奥沙利铂（Oxaliplatin）一线化疗效果的关系。方法：99例晚期大肠癌患者化疗前取静脉血并提取DNA，以实时荧光定量PCR法对ERCC1 Asn118Asn进行SNP分型。患者接受奥沙利铂化疗，观察至疾病进展时间（1TrP）及疾病控制率（DCR），比较不同基因型与化疗效果的关系。结果：ERCC1 Asn118Asn基因位点在本研究人群中的突变频率C／C为50．51％，C／T为41．41％，T／T为8．08％。其中62例Ⅳ期大肠癌患者疾病控制率为54．84％，C／C与C／T＋T／T基因型在疾病控制组（CR＋PR＋SD）和未控组（PD）之间分布差异有统计学意义（OR：3．764，95％CI：1．310～10．813）。37例Ⅲ期大肠癌患者中位DFS为7个月（95％CI：5．483～9．653），C／C型患者中位DFS为10个月（95％CI：8．215～11．642），而C／T＋T／T型患者为5个月（95％CI：4．663～7．267），两者差异显著（Log—rank X^2=12．864，OR=2．237，P〈0．01）。结论：ERCC1 Asn118Asn基因多态性与中国汉族晚期大肠癌患者接受奥沙利铂一线化疗后的临床效果有关。     

晚期大肠癌患者外周血ERCC1基因多态性与奥沙利铂化疗疗效相关性的研究

目的：研究ERCC1 Asn118Asn（C→T，rs11615）多态性与中国汉族晚期大肠癌患者对奥沙利铂（Oxaliplatin）一线化疗效果的关系。方法：99例晚期大肠癌患者化疗前取静脉血并提取DNA，以实时荧光定量PCR法对ERCC1 Asn118Asn进行SNP分型。患者接受奥沙利铂化疗，观察至疾病进展时间（1TrP）及疾病控制率（DCR），比较不同基因型与化疗效果的关系。结果：ERCC1 Asn118Asn基因位点在本研究人群中的突变频率C／C为50．51％，C／T为41．41％，T／T为8．08％。其中62例Ⅳ期大肠癌患者疾病控制率为54．84％，C／C与C／T＋T／T基因型在疾病控制组（CR＋PR＋SD）和未控组（PD）之间分布差异有统计学意义（OR：3．764，95％CI：1．310～10．813）。37例Ⅲ期大肠癌患者中位DFS为7个月（95％CI：5．483～9．653），C／C型患者中位DFS为10个月（95％CI：8．215～11．642），而C／T＋T／T型患者为5个月（95％CI：4．663～7．267），两者差异显著（Log—rank X^2=12．864，OR=2．237，P〈0．01）。结论：ERCC1 Asn118Asn基因多态性与中国汉族晚期大肠癌患者接受奥沙利铂一线化疗后的临床效果有关。     

晚期大肠癌患者血清XRCC1基因多态性与奥沙利铂化疗疗效相关性的研究

目的:探讨XRCC1Arg399Gln(G→)基因多态性与晚期大肠癌对以臭沙利铂为主的化疗敏感性的关系.方法:晚期大肠癌患者99例(Ⅲ期37例,Ⅳ期62例),以奥沙利铂为主的方案化疗,62例Ⅳ期患者2～3个周期后进行临床疗效评价,99例患者统计至疾病进展时间(TTP).应用TaqMan-MGB探针等位基因分型技术进行基因分型.结果:52.53%为G/G,9.09%为A/A,38.38%为G/A.Ⅳ期62例患者化疗后临床获益率(CR PR SD)为54.84%,G/G、G/A A/A在化疗敏感组与不敏感组中的分布具有统计学意义(x2=6.513,P=0.21;OR=3.870,95%CI=1.341～11.172,P=0.012).99例患者中位TTP 7个月,G/G基因型10个月,G/A A/A基因型5个月,两者比较差异有统计学意义,x2=29.20,P<0.01.结论:XRCC1 Arg399Gin基因多态性可能与晚期大肠癌患者对奥沙利铂化疗的敏感性与生存时间相关.     

ERCC 1基因多态性在胃癌中的研究

作为核酸损伤修复过程的重要成员,切除修复交叉互补基因1（ERCC1）的表达情况与胃癌的发生发展密切相关。研究发现,ERCC1基因的多态性会改变该基因自身的表达,并对铂类化疗药物的敏感性和疗效产生影响,通过对基因多态性的检测可以指导胃癌患者的个体化化疗。     

晚期胃癌患者外周血ＥＲＣＣ１基因多态性与奥沙利铂疗效相关性的研究

目的：探讨切除修复交叉互补基因１（ＥＲＣＣ１）Ａｓｎ１１８Ａｓｎ（Ｃ→Ｔ,ｒｓ１１６１５）多态性与中国汉族晚期胃癌患者对奥沙利铂一线化疗效果的关系。方法：７１例晚期胃癌患者化疗前抽静脉血提取ＤＮＡ,以ｒｅａｌ-ｔｉｍｅＰＣＲ法对ＥＲＣＣ１基因进行ＳＮＰ分型。患者接受奥沙利铂化疗,观察疾病控制率（ＣＲ＋ＰＲ＋ＳＤ）及至肿瘤进展时间（ＴＴＰ）,并比较不同基因型与疗效的关系。结果：４７例Ⅳ期胃癌患者可以评价化疗疗效,疾病控制率为５５.３％,Ｃ／Ｃ与Ｃ／Ｔ＋Ｔ／Ｔ基因型在疾病控制组和无效组之间分布无统计学意义（χ２＝2.７５５,Ｐ＝０.０９７）。７１例晚期胃癌患者中位ＴＴＰ为６个月,Ｃ／Ｔ＋Ｔ／Ｔ基因型患者其中位ＴＴＰ为５个月,与纯合基因型Ｃ／Ｃ中位ＴＴＰ６个月相比,差异无统计学意义（ＯＲ＝０.６１５,Ｐ＝０.０７１）。其中２４例Ⅲ期胃癌患者中位ＴＴＰ为８个月,Ｃ／Ｔ＋Ｔ／Ｔ型患者中位ＴＴＰ为７个月,与纯合基因型Ｃ／Ｃ中位ＴＴＰ８个月相比,差异无统计学意义（ＯＲ＝０.３９７,Ｐ＝０.１１１）。结论：ＥＲＣＣ１Ａｓｎ１１８Ａｓｎ基因多态性与中国汉族晚期胃癌患者接受奥沙利铂一线化疗后的临床效果有相关趋势,需要进一步观察论证。     

ERCC1基因多态性与食管癌的研究进展

核苷酸切除修复交叉互补基因1(excision repair cross complementation 1,ERCC1)是核苷酸切除修复(NER)系统的关键酶,其表达情况与食管癌的发生发展及铂类耐药有关.检测ERCC1基因多态性有助于制定食管癌个体化治疗.     

ERCC1基因多态性与胃癌发生发展的关系

  目的  本研究拟探讨切除修复互补交叉基因1(ERCC1)3个单核苷多态性(SNPs)与胃癌发生发展的关系。  方法  选取2007年1月至2009年12月福建地区组织学确诊的452例胃癌患者和469例健康体检人群, 利用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)方法进行rs11615 T > C、rs2298881 C > A和rs3212930 T > C、3个位点多态性检测, 以评估ERCC1不同基因型与胃癌发病风险和病理特征的关系。  结果  rs11615 T > C位点携带C等位基因的个体患胃癌的风险较野生纯合基因型降低一半(OR=0.49;95%CI: 0.52~0.47;P=0.01), 但是罹患弥漫型胃癌的风险上升1.68倍, 预后更差(OR=1.68;95%CI: 0.76~0.61;P=0.048)。单体型分析显示, 基于这3个位点的单体型C-C-C降低了体患胃癌的风险(OR=0.729;95%CI: 0.531~1.001;P=0.0499), 而单体型T-C-T则增加了胃癌患病的风险(OR=1.321;95%CI: 1.063~1.641;P=0.0118)。  结论  ERCC1基因rs11615 T > C位点多态性与胃癌发生发展密切相关, 携带该基因2种不同单体型的个体在胃癌患病风险上存在差异。      

shRNA靶向沉默ST8SIA4基因表达对乳腺癌细胞侵袭和迁移的影响

目的 探讨短发夹RNA（shRNA）靶向沉默α2, 8-唾液酸转移酶4（ST8SIA4）表达对乳腺癌BT549细胞侵袭、迁移的影响。方法 采用实时荧光定量PCR（QPCR）和Western blotting检测乳腺癌BT549细胞和乳腺正常上皮MCF-10A细胞中ST8SIA4 mRNA和蛋白水平。采用LipofectamineTM2000向BT549细胞随机转染成功构建的靶向沉默ST8SIA4表达的shRNA载体片段（沉默组）或无义shRNA片段（对照组）,转染48 h后采用QPCR检测ST8SIA4 mRNA水平,Western blotting检测ST8SIA4、磷酸化丝／苏氨酸蛋白激酶（p-Akt）、神经纤毛蛋白2（NRP2）和肿瘤坏死因子-α（TNF-α）的表达情况,Transwell实验和划痕实验分别检测细胞的侵袭和迁移能力。结果 与MCF-10A细胞相比,BT549细胞的ST8SIA4 mRNA和蛋白水平均升高（P＜0.05）。转染48 h后,沉默组的ST8SIA4 mRNA和蛋白水平分别为0.19±0.02和0.13±0.02,均低于对照组的0.98±0.11和0.52±0.05（P＜0.05）;沉默组的侵袭细胞数、迁移率及p-Akt、NRP2和TNF-α蛋白相对表达量分别为（39.5±4.2）个、（16.9±1.3）％、0.22±0.03、0.31±0.04和0.27±0.06,均低于对照组的（91.3±8.5）个、（38.4±3.9）％、0.58±0.07、0.55±0.05和0.46±0.05,差异有统计学意义（P＜0.05）。结论 ST8SIA4基因在乳腺癌BT549细胞中高表达,沉默其表达可抑制侵袭和迁移过程,可能与上调p-Akt、NRP2和TNF-α蛋白表达有关。     

Impact of ERCC1 expression on treatment outcome in small-cell lung cancer patients treated with platinum-based chemotherapy

IntroductionThe excision repair cross-complementing 1 (ERCC1) protein is an extensively investigated molecular marker because it may decrease sensitivity to platinum-based chemotherapy. Low ERCC1 expression has already been correlated with better treatment efficacy in non-small-cell lung cancer patients treated with platinum-based chemotherapy. However, the data on a prognostic and/or predictive value of ERCC1 in small-cell lung cancer (SCLC) are still very limited.MethodsThis retrospective pilot study evaluated the impact of ERCC1 expression levels on response to first-line platinum-based chemotherapy with or without radiotherapy and survival outcomes of 77 SCLC patients. ERCC1 protein expression was determined immunohistochemically in primary tumour tissue.ResultsERCC1 protein expression was positive in 40/77 (51.9%) of our patients. No significant association was found between ERCC1 protein expression and response rate to first-line platinum-based chemotherapy, progression-free survival (PFS), or overall survival (OS), either in the overall population or in patients stratified by disease stage.ConclusionsIn our limited group of 77 SCLC patients, ERCC1 protein expression was not found to correlate with either response rate to platinum-based chemotherapy or survival outcomes. Multi-centric prospective trials using a validated method of ERCC1 determination are mandatory in order to obtain a definitive answer on the predictive value of ERCC1 in SCLC.     

ERCC1 codon 118 polymorphism is a useful prognostic marker in patients with pancreatic cancer treated with platinum-based chemotherapy

Excision repair cross-complementation 1 (ERCC1) has been reported to play a major role in the response to platinum-based therapies. It has recently been proposed that a synonymous polymorphism at codon 118 converting a common codon usage (AAC) to an infrequent one (AAT) may impair ERCC1 translation and to affect the response to cisplatin chemotherapy. We analyzed the association between this polymorphism and clinical outcome in 67 pancreatic cancer patients treated with cisplatin and S-1 or with S-1 alone. ERCC1 codon 118 polymorphism was analyzed using PCR-RFLP. Thirty-nine of the patients (58.2%) were homozygous for AAC codon, 7 (10.4%) were homozygous for AAT codon, and 21 (31.3%) were heterozygous. Among those treated with cisplatin and S-1, no significant difference in objective response rate was observed between genotypes. However, the patients with one or two AAT codons had a significantly longer progression-free survival (PFS) and overall survival (OS) than those homozygous for AAC allele (PFS: 338 days vs 106 days, p=0.006, OS: 763 days vs 415 days, p=0.030). In contrast, no significant difference in PFS or OS was observed between genotypes among the patients treated with S-1 alone. ERCC1 polymorphism may be a useful prognostic marker in patients with pancreatic cancer treated with platinum-based chemotherapy.     

ERCC1 and BRCA1 mRNA expressions are associated with clinical outcome of non-small cell lung cancer treated with platinum-based chemotherapy

We conducted a perspective study to investigate whether the expression of excision repair cross-complementing 1 (ERCC1), xeroderma pigmentosum group G (XPG), breast cancer 1 (BRCA1), and ribonucleotide reductase M1 (RRM1) is correlated with clinical outcome of non-small cell lung cancer (NSCLC). Patients with histologically confirmed inoperable stages IIIB and IV NSCLC were collected and followed up until January 2012. Relative cDNA quantification for ERCC1, XPG, BRCA1, and RRM1 was performed using a fluorescence-based, real-time detection method. Cox regression analysis indicated that a high level of ERCC1 was associated with shorter overall survival (OS) and progression-free survival (PFS) times when compared with low expression, with adjusted hazard ratios (HRs) (95 % confidence interval (CI)) of 2.25 (1.18–4.39) and 2.63 (1.33–5.25), respectively. High expression of BRCA1 was correlated with shorter OS and PFS times when compared with low expression, and the adjusted HRs (95 % CI) were 3.29 (1.72–6.39) and 5.94 (2.80–13.06), respectively. Moreover, we found a significant correlation between BRCA1 expression and age (χ ²?=?4.14, P?=?0.04) and stage (χ ²?=?5.26, P?=?0.02). Our results suggest that ERCC1 and BRCA1 mRNA expressions are associated with PFS and OS in advanced NSCLC patients treated with platinum-based chemotherapy.     

Association of RRM1 -37A>C polymorphism with clinical outcome in colorectal cancer patients treated with gemcitabine-based chemotherapy

Background

To investigate whether single nucleotide polymorphisms (SNPs) in gemcitabine (GMB) metabolism genes were associated with clinical outcome in pre-treated metastatic colorectal cancer (mCRC) patients.

Patients and methods

SNPs of hCNT1, hENT1, CDA, dCTD and RRM1 genes were evaluated in 95 mCRC patients and detected using TaqMan genotyping assays. Association of genotypes with overall response rate (ORR), time to progression (TTP) and overall survival (OS) was tested by univariate and multivariate analysis. RRM1 -37A>C polymorphism was correlated with GMB IC50 value and with the RRM1 gene expression level in CRC cell lines.

Results

The ORR was 38.9%. The median TTP and OS were 4 and 14.3 months, respectively. By multivariate analysis, patients carrying the RRM1 -37CC genotype or the CDA A-76 C-containing allele had a significantly higher likelihood of achieving a tumour response. RRM1 -37A>C polymorphism remained associated with clinical efficacy (TTP). In vitro experiments, in CRC cell lines, showed that the RRM1 A-37C genotype was associated with the levels of RRM1 expression and with GMB IC50 values. Finally, the down-regulation of RRM1 with a specific siRNA strongly influenced GMB sensitivity.

Conclusion

RRM1 -37A>C polymorphism may represent a useful biomarker to select mCRC patients most likely to benefit from GMB-based salvage therapy.     

Clinical outcome for EML4-ALK-positive patients with advanced non-small-cell lung cancer treated with first-line platinum-based chemotherapy

BackgroundThe EML4-ALK fusion oncogene represents a recently identified molecular target in a subset of patients with non-small-cell lung cancer (NSCLC). Limited data have been available, however, on the outcome of first-line platinum-based chemotherapy in patients with EML4-ALK-positive advanced NSCLC who have not been treated with an ALK kinase inhibitor.Patients and methodsThe efficacy of platinum-based chemotherapy was compared between patients with advanced nonsquamous NSCLC who harbor EML4-ALK and those who harbor EGFR mutations and those with neither molecular abnormality.ResultsAmong 200 patients with advanced nonsquamous NSCLC, 18 (9.0%) were positive for EML4-ALK, 31 (15.5%) harbored EGFR mutations, and 151 (75.5%) were wild type for both abnormalities. Platinum-based combination chemotherapy showed similar efficacies in the EML4-ALK, EGFR mutation, and wild-type cohorts in terms of response rate and progression-free survival, and overall survival in the EML4-ALK cohort closely resembled that in the wild-type cohort. Within the EML4-ALK cohort, patients with variants 1 or 3 of the fusion gene were predominant and did not appear to differ in their sensitivity to the platinum-based regimens.ConclusionPatients with EML4-ALK-positive advanced NSCLC manifest an aggressive clinical course similar to that of those with wild-type tumors if the effective targeted therapy is not instituted.     

Relationship between systemic 5-FU passage and response in colorectal cancer patients treated with intrahepatic chemotherapy

Summary The study described herein was conducted to analyze the relationship between tumor exposure to 5-FU and clinical response. Six patients were placed on continuous 5-day intrahepatic 5-FU chemotherapy for colorectal cancer metastasized to the liver. The starting dose was 600–800 mg/m² per day; cycles were repeated at 4-week intervals. The 5-FU dose was increased by 250 mg/day at each cycle. All six patients received 3 or more cycles, for a total of 37 cycles. Response was evaluated after each cycle by ultrasonography or computed tomography (CT). Pharmacokinetic data revealed a high individual cycle-to-cycle variability for all six patients in the 5-FU area under the curve (AUC day 1 to day 5) corrected for the dose. These variations in drug biodisposition, reflecting hepatic 5-FU uptake, were significantly related to measurable modifications in the tumor mass in 71% of cycles. The correlation between the reduction in local drug exposure and tumor regrowth was better than that between the increase in local drug exposure and tumor reduction. These findings constitute an original illustration in humans of the experimental concept of the drug exposure/tumor response relationship for 5-FU.     

ERCC1和XPD基因SNPs与NSCLC含铂类药物化疗敏感相关性分析

目的:探讨切除修复交叉互补基因1(ERCC1)第118位密码子(C118T)和着色性干皮病基因D(XPD)第751位密码子(Lys751Gln)的单核苷酸多态性(SNPs),与晚期非小细胞肺癌(NSCLC)患者对含铂类药物化疗疗效的关系。方法:聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术检测78例NSCLC患者的ERCC1(C118T)和XPD(Lys751Gln)基因型。结果:ERCC1(C118T)SNPs与铂类化疗疗效有关,P<0.05。C/C型化疗有效者7例(8.9%),C/T型为11例(14.1%),T/T型2例(2.6%);C/C化疗无效41例(52.6%),C/T型17例(21.8%),T/T型0;两组差异有统计学意义,P=0.003。T等位基因(T/T+C/T)型与C等位基因(C/C)型的化疗疗效差异亦有统计学意义,P=0.005,比值比(OR)=0.223,95%的可信区间(CI)为0.076～0.657;XPD(Lys751gln)SNPs与含铂类药物化疗疗效无关。ERCC1(C118T)、XPD(Lys751Gln)SNPs分布与年龄、性别、吸烟史、临床分期、体力状况评分(ECOG)和病理类型无关,P>0.05。结论:ERCC1(C118T)含有T等位基因,可作为预测晚期NSCLC患者铂类药物化疗敏感性的预测指标。