多靶标检测平台分析肺腺癌患者不同分化程度中ERCC1、TYMS、RRM1、TUBB3基因表达及意义

目的:通过一种mRNA表达水平多指标检测技术分析ERCC1、TYMS、TUBB3、RRM1在非小细胞肺腺癌(NSCLS)患者人群中的表达,探索基因表达与腺癌病理分化程度之间的相关性,并探讨其临床意义,为个体化用药临床指导提供基础。方法:应用分支DNA-液相芯片基因检测技术检测30例肺腺癌患者上述基因的mRNA水平表达,使用SPSS软件统计分析基因表达与腺癌分化程度之间的相关性。结果:ERCC1表达与腺癌分化程度有显著相关性(P<0.05),除此之外,其它靶标与腺癌分化程度之间并无显著相关性(P>0.05)。结论:ERCC1表达可能与腺癌分化程度之间具有相关性,腺癌中分化的患者ERCC1平均表达水平较低,低分化的患者ERCC1平均表达水平较高;同时,当ERCC1低表达时(24.1%),其他三个指标同时低表达的比例更低(分别为10.3%、6.9%、6.9%),因此确定用药方案之前,参考病理特征及靶标检测还是非常必要的。     

非小细胞肺癌组织中 EGFR 基因突变与 ERCC1 mRNA 表达的关系

目的：探讨非小细胞肺癌中EGFR基因突变与ERCC1 mRNA 表达之间的关系。 方法：收集NSCLC患者病例样本41例,应用ARMS方法检测EGFR基因突变,采用RT-PCR方法检测ERCC1 mRNA 表达,应用Spearman相关检验对EGFR突变状态与ERCC1 mRNA 的表达进行相关性分析。 结果：在41例患者中,EGFR突变21例,ERCC1 mRNA 高、中、低表达率分别为19.1%(4/21)、57.1%(12/21)和23.8%(5/21),EGFR基因突变与ERCC1 mRNA 表达显著相关(P < 0.001)。 结论: NSCLC EGFR基因突变与ERCC1 mRNA 表达具有显著相关性。     

Coexistence of EGFR with KRAS,or BRAF,or PIK3CA somatic mutations in lung cancer: a comprehensive mutation profiling from 5125 Chinese cohorts

Background:

Determining the somatic mutations of epidermal growth factor receptor (EGFR)-pathway networks is the key to effective treatment for non-small cell lung cancer (NSCLC) with tyrosine kinase inhibitors (TKIs).The somatic mutation frequencies and their association with gender, smoking history and histology was analysed and reported in this study.

Methods:

Five thousand one hundred and twenty-five NSCLC patients'' pathology samples were collected, and EGFR, KRAS, BRAF and PIK3CA mutations were detected by multiplex testing. The mutation status of EGFR, KRAS, BRAF and PIK3CA and their association with gender, age, smoking history and histological type were evaluated by appropriate statistical analysis.

Results:

EGFR, KRAS, BRAF and PIK3CA mutation rates revealed 36.2%, 8.4%, 0.5% and 3.3%, respectively, across the 5125 pathology samples. For the first time, evidence of KRAS mutations were detected in two female, non-smoking patients, age 5 and 14, with NSCLC. Furthermore, we identified 153 double and coexisting mutations and 7 triple mutations. Interestingly, the second drug-resistant mutations, T790M or E545K, were found in 44 samples from patients who had never received TKI treatments.

Conclusions:

EGFR exons 19, 20 and 21, and BRAF mutations tend to happen in females and non-smokers, whereas KRAS mutations were more inclined to males and smokers. Activating and resistant mutations to EGFR-TKI drugs can coexist and ‘second drug-resistant mutations'', T790M or E545K, may be primary mutations in some patients. These results will help oncologists to decide candidates for mutation testing and EGFR-TKI treatment.     

原发小细胞肺癌中BRAF/KRAS以及PIK3CA突变的基因特征及临床特征

目的 在中国人群小细胞肺癌(SCLC)标本中检测BRAF/KRAS以及PIK3CA基因突变频率,分析这些基因突变的基因特征和临床特征。方法 2009-2014年共收集557例单纯SCLC患者组织样本。利用双脱氧测序法进行BRAF、KRAS、PIK3CA、NRAS、MEK1基因突变检测。χ²检验分析临床因素与基因突变的相关性,Kaplan-Meier法生存分析,Cox模型多因素预后分析。结果 在557例标本中检测到13例BRAF突变,突变类型包括V600E (n=5)、V600A (n=2)、V600M (n=1)、D594G (n=1)、G464E (n=1)、K601R (n=2)、S605N (n=1)。6例KRAS突变,突变类型包括G12C (n=3)、G12A (n=1)、G12D (n=1)、G13D (n=1)。4例PIK3CA突变,突变类型包括E545G (n=2)、H1047R (n=2)。另外1例NRAS突变(Q61R)和1例MEK1突变(D61Y)。这些突变基因与患者年龄、性别、吸烟状态、临床分期均无相关性。单因素生存分析结果显示基因突变组患者的生存时间比无此类突变者生存时间差,中位生存时间分别为(10.30±0.75)个月(95%CI为8.83~11.77个月)和(12.80±0.54)个月(95%CI为11.74~13.86)(P=0.011)。结论 在单纯SCLC中存在小比例的BRAF/KRAS、PIK3CA基因突变群体,这些基因突变与患者的临床特征无明显统计学相关性,但单因素生存分析显示与患者生存预后呈负相关。     

非小细胞肺癌EGFR、KRAS基因突变与RRM1、TUBB3 mRNA表达水平的相关性

目的:探讨非小细胞肺癌（NSCLC）EGFR、KRAS基因突变与核苷酸还原酶亚基M1（RRM1）、Ⅲ型β微管蛋白（TUBB3）mRNA表达水平的相关性及其临床意义。方法:69例NSCLC组织标本来自解放军空军总医院胸外科2010年6月至2014年10月手术中从肿瘤中心切取。使用xTAG-液相芯片法检测EGFR、KRAS基因突变,分支DNA-液相芯片法检测RRM1、TUBB3 mRNA表达水平。结果:在69例NSCLC组织样本中,28例为EGFR基因突变（40.6%,28/69）,13例为KRAS基因突变（18.8%,13/69）,EGFR基因突变与性别（P=0.005）、病理类型（P=0.036）、吸烟史（P=0.029）相关。KRAS基因突变与性别相关（P=0.007）。TUBB3 mRNA的表达水平与病理类型相关（P=0.008）,RRM1 mRNA的表达水平与患者各临床病理特征无关（P＞0.05）。EGFR基因突变与RRM1、TUBB3 mRNA表达水平无关（P＞0.05）,KRAS基因突变与RRM1 mRNA表达水平无关（P＞0.05）,KRAS突变型患者TUBB3 mRNA表达水平比KRAS野生型患者高（P＜0.05）。结论:在NSCLC患者中,EGFR及KRAS基因的突变状态对评估患者使用吉西他滨及抗微管类药物的疗效有重要意义,尤其在KRAS突变型NSCLC患者中,抗微管类化疗药物的疗效可能不佳,这有利于指导化疗药物方案的制定,促进NSCLC的个体化治疗。     

Combined analysis of mRNA expression of ERCC1, BAG-1, BRCA1, RRM1 and TUBB3 to predict prognosis in patients with non-small cell lung cancer who received adjuvant chemotherapy

Background

The aim of this study was to investigate prognostic value of excision repair cross-complementing 1 (ERCC1), BCL2-associated athanogene (BAG-1), the breast and ovarian cancer susceptibility gene 1 (BRCA1), ribonucleotide reductase subunit M1 (RRM1) and class III β-tubulin (TUBB3) in patients with non-small cell lung cancer (NSCLC) who received platinum- based adjuvant chemotherapy.

Methods

Messenger RNA expressions of these genes were examined in 85 tumor tissues and 34 adjacent tissue samples using semi-quantitative RT-PCR. The expressions of these five genes were analyzed in relation to chemotherapy and progression-free survival (PFS) and overall survival (OS). Seventy-four patients were enrolled into chemotherapy.

Results

Patients with ERCC1 or BAG-1 negative expression had a significantly longer PFS (P = 0.001 and P = 0.001) and OS (P = 0.001 and P = 0.001) than those with positive expression. Patients with negative ERCC1 and BAG-1 expression benefited more from platinum regimen (P = 0.001 and P = 0.002). Patients with BRCA1 negative expression might have a longer OS (P = 0.052), but not PFS (P = 0.088) than those with BRCA1 positive expression. A significant relationship was observed between the mRNA expression of ERCC1 and BAG-1 (P = 0.042). In multivariate analysis, ERCC1 and BAG-1 were significantly favorable factors for PFS (P = 0.018 and P = 0.017) and OS (P = 0.027 and P = 0.022).

Conclusions

ERCC1 and BAG-1 are determinants of survival after surgical treatment of NSCLC, and its mRNA expression in tumor tissues could be used to predict the prognosis of NSCLC treated by platinum.     

mRNA Expression and Clinical Significance of ERCC1, BRCA1, RRM1, TYMS and TUBB3 in Postoperative Patients with Non-Small Cell Lung Cancer

Background: To explore mRNA expression and clinical significance of ERCC1, BRCA1, RRM1, TYMS andTUBB3 genes in tumor tissue of postoperative patients with non-small cell lung cancer (NSCLC). Materialsand Methods: Sixty NSCLC patients undergoing radical operation in our hospital from Nov., 2011 to Jun.,2012 were selected. Plasmid standards of ERCC1, BRCA1, RRM1, TYMS and TUBB3 were established andstandard curves were prepared by SYBR fluorescent real-time quantitative PCR analysis. Samples fromtumor centers were taken to detect mRNA expression of ERCC1, BRCA1, RRM1, TYMS and TUBB3 genesin cancerous tissue during operation. The total mRNA expression quantities were compared according todifferent clinical characteristics. Results: The total expression quantities of 5 genotypes from high to low wereERCC1>RRM1>TUBB3>TYMS>BRCA1 in turn. By pairwise comparisons, other differences showed statisticalsignificance (p<0.05 or p<0.01) except for TYMS and TUBB3 (p>0.05); the low expression rates from high to lowwere ERCC1>TYMS>TUBB3>TUBB3>RRM1>BRCA1 in turn. The expression quantities of BRCA1, RRM1 andTYMS in males, smokers and patients without adenocarcinoma were all significantly higher than that in females,non-smokers and patients with adenocarcinoma, and significant differences were present (p<0.05 or p<0.01).In terms of pathological staging, the expression quantities of BRCA1, RRM1 and TYMS in phases Ⅱa~Ⅱb andⅢa~Ⅲb had a tendency to be greater than in phases Ⅰ and Ⅳ. Conclusions: Resistance to chemotherapy andsensitivity to targeted therapy differ among patients with NSCLC. Differences in gene expression in differentindividuals were also revealed. Only according to personalized detection results can individualized therapeuticregimens be worked out, which is a new direction for oncotherapy.     

ERCC1、RRM1和BRCA1在非小细胞肺癌中的表达及预后意义

目的:探讨DNA修复基因家族成员ERCC1、RRM1和BRCA1在非小细胞肺癌(NSCLC)中的表达及预后意义。方法:应用实时荧光定量PCR技术对32例肺癌及16例癌旁组织中ERCC1、RRM1和BRCA1基因的mRNA进行定量检测。用非参数检验、相关分析、Kap lan-M e ier生存曲线和COX多因素回归分析进行统计分析。结果:NSCLC中ERCC1、RRM1和BRCA1在癌组织内表达量显著高于癌旁组织,且在癌内表达具有正相关性;RRM1在肺鳞癌中高于腺癌,但在不同分期中表达无差异;ERCC1和BRCA1在不同病理类型和分期中表达均无差异;RRM1和BRCA1高表达组的生存期明显长于低表达组;COX多因素回归分析示RRM1表达是影响本组患者预后的独立因素。结论:NSCLC中,ERCC1、RRM1和BRCA1在肺癌组织中的表达显著高于癌旁组织,RRM1和BRCA1高表达组的生存期长于低表达组。RRM1和BRCA1可作为判断预后的一种指标。     

ERCC1、TYMS、RRM1、TUBB3表达与非小细胞肺癌化疗疗效的相关性和意义

[目的]探讨非小细胞肺癌(NSCLC)术后标本中ERCC1、TYMS、RRM1、TUBB3表达与化疗疗效的相关性.[方法]回顾性分析我院病理确诊的72例NSCLC患者手术切除的肿瘤标本,通过分支DNA-液相芯片法,检测ERCC1、TYMS、RRM1、TUBB3基因mRNA表达水平.[结果]ERCC1、TYMS、RRM1、TUBB3阳性表达率分别为38.9％ (28/72)、62.5％(15/72)、55.6％ (46/72)和47.2％(34/72);4者表达与年龄、性别、吸烟、组织学类型、TNM分期、淋巴结转移均无关(P＞0.05),TYMS、TUBB3在高、中分化组中的阳性表达率显著低于低分化组(P=0.003,P＜0.001).ERCC1、TYMS、RRM1和TUBB3阳性表达的化疗患者1年生存率均显著低于阴性表达者(61.5％vs95.5％,P=0.048;53.5％vs 94.9％,P=0.035;58.6％vs 92.9％,P=0.039;47.8％vs 97.4％,P=0.014).[结论]ERCC1、TYMS、RRM1、TUBB3阴性表达者化疗生存率高.ERCC1、TYMS、RRM1、TUBB3阴性表达可作为NSCLC患者临床化疗受益的指标.     

ERCC1, RRM1 and BRCA1 mRNA expression levels and clinical outcome of advanced non-small cell lung cancer

This study was to investigate whether the expressions of DNA repair genes ERCC1 (excision repair cross complementing 1), RRM1 (ribonucleotide reductase subunit M1) and BRCA1 (breast cancer 1) affected clinical outcome in patients with NSCLC. Patients with stage IIIb/IV NSCLC were given platinum-based chemotherapy. Messenger RNA expression levels of ERCC1, BRCA1 and RRM1 were determined by real-time polymerase chain reaction with TaqMan probes in the tumor. The relationship between these three genes with chemoresponse and overall survival was analyzed in this study. Eighty-five patients (median age 59, range 30–78) were enrolled into the study. Median overall survival (OS) was 13 months (range 10.8–15.2). Time to progression (TTP) was 6.1 months (range 5.5–6.7). Patients with low ERCC1 expression benefited more from a platinum-containing regimen (P = 0.094). Patients with low RRM1 expression benefited more from a gemcitabine-containing regimen. Patients with high BRCA1 expression benefited more from an anti-tubulin-containing regimen (P = 0.046). Partial response rate was 42.4%. A statistically significant difference in OS was seen in patients with low ERCC1 levels compared to patients with high ERCC1 ones. (16.5 vs. 10.0 months, P = 0.045). A significant relationship was observed between the expression of ERCC1 and BRCA1 and TTP (6.5 vs. 5.1 months, P = 0.001, 5.2 vs. 6.5, P = 0.019, respectively). High expression of BRCA1 was associated with better survival in the anti-tubulin-containing regimen subgroup (8.7 vs. 13.0, P = 0.035). ERCC1, RRM1 and BRCA1 are promising predictive and prognostic biomarkers in advanced non-small cell lung cancer.     

非小细胞肺癌EGFR突变与ERCC1和BRCA1表达相关性的研究

目的:探讨非小细胞肺癌(NSCLC) EGFR突变与ERCC1、BRCA1表达的相关性.方法:收集符合入组条件的NSCLC患者151例,应用RT-PCR方法检测EGFR突变,采用免疫组化DAB法检测ERCC1、BRCA1蛋白表达,应用Fisher精确检验对EGFR突变状态与ERCC1、BRCA1的表达进行相关性分析.结果:151例患者中,EGFR突变37例,ERCC1低、中、高表达率分别为43.2％ (16/37)、54.1％(20/37)和2.7％(1/37),EGFR突变与ERCC1表达有相关性,P=0.009;BRCA1低、中、高表达率分别为43.2％(16/37)、48.6％0(18/37)和8.1％(3/37),EGFR突变与BRCA1表达无相关性,P=0.323.结论:NSCLC EGFR突变与ERCC1蛋白表达有相关性,与BRCA1蛋白表达无相关性.     

The prognostic role of KRAS,BRAF, PIK3CA and PTEN in colorectal cancer

Background

Mutations in KRAS, BRAF, PIK3CA and PTEN expression have been in focus to predict the effect of epidermal growth factor receptor-blocking therapy in colorectal cancer (CRC). Here, information on these four aberrations was collected and combined to a Quadruple index and used to evaluate the prognostic role of these factors in CRC.

Patients

We analysed the mutation status in KRAS, BRAF and PIK3CA and PTEN expression in two separate CRC cohorts, Northern Sweden Health Disease Study (NSHDS; n=197) and Colorectal Cancer in Umeå Study (CRUMS; n=414). A Quadruple index was created, where Quadruple index positivity specifies cases with any aberration in KRAS, BRAF, PIK3CA or PTEN expression.

Results

Quadruple index positive tumours had a worse prognosis, significant in the NSHDS but not in the CRUMS cohort (NSHDS; P=0.003 and CRUMS; P=0.230) in univariate analyses but significance was lost in multivariate analyses. When analysing each gene separately, only BRAF was of prognostic significance in the NSHDS cohort (multivariate HR 2.00, 95% CI: 1.16–3.43) and KRAS was of prognostic significance in the CRUMS cohort (multivariate HR 1.48, 95% CI: 1.02–2.16). Aberrations in PIK3CA and PTEN did not add significant prognostic information.

Conclusions

Our results suggest that establishment of molecular subgroups based on KRAS and BRAF mutation status is important and should be considered in future prognostic studies in CRC.     

非小细胞肺癌表皮生长因子受体基因拷贝数与核苷酸切除修复交叉互补基因1和乳腺癌易感基因1的表达及其相关性分析

目的 探讨表皮生长因子受体(EGFR)基因拷贝数与铂类耐药相关基因核苷酸切除修复交叉互补基因1(ERCC1)和乳腺癌易感基因1(BRCA1)在非小细胞肺癌(NSCLC)中的表达情况,以及三者之间的相关性.方法 用荧光原位杂交(FISH)技术检测EGFR基因的扩增情况,应用免疫组化SP法检测132例NSCLC患者标本ERCC1和BRCA1蛋白的表达,并进一步分析EGFR基因拷贝数、ERCC1和BRCA1的蛋白表达与患者临床病理特征的关系.结果 EGFR基因FISH阳性率为24.1% (40/166).女性的扩增阳性率高于男性(31.9%和18.6%,P=0.048);无吸烟史的患者扩增阳性率明显高于有吸烟史的患者(32.8%和16.7%,P=0.045);基因拷贝数增加与患者的年龄、临床分期、病理类型及有无淋巴结或远处转移均无关.ERCC1蛋白表达阳性率为45.5% (60/132),在不同的病理类型分布比较,差异有统计学意义(P=0.046),与患者的性别、年龄、临床分期、有无淋巴结或远处转移及吸烟状况均无关.BRCA1蛋白表达阳性率为35.1%(46/131),与患者的性别、年龄、临床分期、病理类型、淋巴结或远处转移及吸烟状况无关.ERCC1与BRCA1的蛋白表达呈中等相关(r=0.449,P＜0.001),与EGFR扩增无相关性(P=0.785);BRCA1表达与EGFR基因拷贝数状态无相关性(P=0.143).结论 在肺癌的个体化治疗中,检测EGFR基因拷贝数、ERCC1和BRCA1的表达对靶向治疗和铂类药物化疗的选择具有重要的临床指导意义.

Abstract：

Objective To evaluate the expression of epidermal growth factor receptor (EGFR) gene copy number and the expression of ERCC1 and BRCA1 proteins in patients with non-small-cell lung cancer (NSCLC) and the correlation between them. Methods The status of EGFR gene copy number was determined by in situ hybridization (FISH), and the expression of ERCC1 and BRCC1 proteins was examined by immunohistochemistry (IHC). The relationship of EGFR gene copy number with the expression of ERCC1 and BRCA1 and the clinical pathologic features were analyzed. Results FISH-positive EGFR expression was identified in 40 of 166 samples (24.1%). More FISH-positive EGFR in the female than male patients (31.9% vs. 18.6%, P=0.048), and non-smoker than smoker (32.8% vs. 16.7%, P=0.045). FISH-positive EGFR was not associated with age, pathological type, clinical stage and metestasis status (P＞0.05). The expression of ERCC1 protein was identified in 60 of 132 samples (45.5%). The expression of ERCC1 protein varied significantly in tumors of different pathological types (P=0.046), but not associated with age, gender, clinical stage, metestatic status and smoking status (P＞0.05). The expression of BRCA1 protein was identified in 46 of 131 samples (35.1%). The expression of BRCA1 was not associated with age gender, pathological type, clinical stage, metestatic ststus and smoking status (P＞0.05). There was a moderate correlation between the expressions of ERCC1 and BRCA1 (r=0.449, P＜0.001), but EGFR gene copy number was not correlated with the expression of ERCC1 or BRCA1 protein. Conclusions FISH-positive EGFR expression is associated with gender and smoking status, but not correlated with the expression of ERCC1 and BRCA1 proteins. There is a moderate correlation between the expressions of ERCC1 and BRCA1.     

ERCC1、RRM1基因表达与非小细胞肺癌患者预后的相关性

目的探讨ERCC1、RRM1基因蛋门表达对Ⅰ～Ⅲa期非小细胞肺癌患者预后的影响。方法采用免疫组化法检测Ⅰ～Ⅲa期非小细胞肺癌患者ERCC1、RRM1蛋白表达情况;采用生存分析曲线分析其对生俘预后的影响。结果ERCCl、RRMl蛋白表达与患者性别、年龄、病珲类型和TNM分期等莘异均无统计学意义（P〉0．05）。在TNM分期中,Ⅰa期ERCCl、RRM1阳性表达组生存预后较好,提示两种基因蛋白的阳性表达在18期患者中是一种保护因素。存IhⅢa期患者中ERCC1表达阴性组可以从含铂化疗方案中获益,可获得生存优势;RRMI表达阴性组对化疗药物吉西他滨敏感,可获得较长的生存优势。在Ⅰ～Ⅲa期患者中ERCC1与RRM1的表达呈正相关,差异有统计学意义（P〈0．05）。结论检测Ⅰ～Ⅲa期非小细胞肺癌ERCCI及RRM1蛋白表达可以预测患者的治疗疗效及预后,使患者从个体化治疗中获益。     

Optimization of patient selection for EGFR-TKIs in advanced non-small cell lung cancer by combined analysis of KRAS, PIK3CA, MET, and non-sensitizing EGFR mutations

Background

We present a comprehensive analysis of KRAS, PIK3CA, MET, and non-sensitizing EGFR mutations in advanced non-small cell lung cancer (NSCLC) patients treated with tyrosine kinase inhibitors (TKIs), with the aim of clarifying the relative contribution of these molecular alterations to resistance.

Patients and methods

One hundred and sixty-six patients with advanced NSCLC treated with EGFR-TKIs with available archival tissue specimens were included. EGFR (exons 18–21), KRAS (exons 2, 3), PIK3CA (exons 9, 20), and MET (exons 14, 15) mutations were analyzed using PCR-based sequencing. Among all the mutations evaluated, only KRAS, PIK3CA, MET, and non-sensitizing EGFR mutations, defined as “TKI non-sensitizing mutations” were used for response, time to progression (TTP), and overall survival (OS) analysis.

Results

TKI non-sensitizing mutations were associated with disease progression (p?=?0.001), shorter TTP (p?p?=?0.03). Cox’s multivariate analysis including histology and performance status showed that TKI non-sensitizing mutations were independent factors for shorter TTP (p?p?=?0.01).

Conclusions

When KRAS, PIK3CA, MET, and non-sensitizing EGFR mutations are concomitant, up to 96.0% of NSCLC patients unlikely to respond to TKIs can be identified, and they represented independent negative prognostic factors. Comprehensive molecular dissection of EGFR signaling pathways should be considered to select advanced NSCLC patients for TKIs therapies.     

Molecular spectrum of KRAS,BRAF, and PIK3CA gene mutation: determination of frequency,distribution pattern in Indian colorectal carcinoma

Molecular evaluation of KRAS, BRAF, and PIK3CA mutation has become an important part in colorectal carcinoma evaluation, and their alterations may determine the therapeutic response to anti-EGFR therapy. The current study demonstrates the evaluation of KRAS, BRAF, and PIK3CA mutation using direct sequencing in 204 samples. The frequency of KRAS, BRAF, and PIK3CA mutations was 23.5, 9.8, and 5.9 %, respectively. Five different substitution mutations at KRAS codon 12 (G12S, G12D, G12A, G12V, and G12C) and one substitution type at codon 13 (G13D) were observed. KRAS mutations were significantly higher in patients who were >50 years, and were associated with moderate/poorly differentiated tumors and adenocarcinomas. All mutations in BRAF gene were of V600E type, which were frequent in patients who were ≤50 years. Unlike KRAS mutations, BRAF mutations were more frequent in well-differentiated tumors and right-sided tumors. PIK3CA–E545K was the most recurrent mutation while other mutations detected were T544I, Q546R, H1047R, G1049S, and D1056N. No significant association of PIK3CA mutation with age, tumor differentiation, location, and other parameters was noted. No concomitant mutation of KRAS and BRAF mutations was observed, while, interestingly, five cases showed concurrent mutation of KRAS and PIK3CA mutations. In conclusion, to our knowledge, this is the first study to evaluate the PIK3CA mutation in Indian CRC patients. The frequency of KRAS, BRAF, and PIK3CA was similar to worldwide reports. Furthermore, identification of molecular markers has unique strengths, and can provide insights into the pathogenic process and help optimize personalized prevention and therapy.     

核苷酸切除修复交叉互补基因1 mRNA表达与非小细胞肺癌铂类化疗患者预后的相关性

目的 探讨核苷酸切除修复交叉互补基因1(ERCC1)mRNA表达与非小细胞肺癌(NSCLC)铂类化疗患者临床病理特征的关系及其预后意义.方法 采用实时荧光定量逆转录聚合酶链反应(RT-PCR)方法,检测NSCLC石蜡包埋组织中ERCC1 mRNA的表达水平,并比较其表达水平与NSCLC铂类化疗患者临床病理特征和生存时间之间的关系.结果 61例NSCLC患者中,ERCC1 mRNA的中位表达量为0.48.ERCC1 mRNA表达与NSCLC患者临床病理特征无关.ERCC1mRNA低表达(<0.35)患者经铂类药物化疗后的无进展生存时间为14.3个月,而高表达者为8.0个月,差异有统计学意义(P=0.028).ERCC1 mRNA低表达(<0.28)患者的总生存时间为28.4个月,而高表达者为12.9个月,差异有统计学意义(P=0.0064).Cox多因素回归分析显示,ERCC1 mRNA表达水平是影响NSCLC患者预后的独立因素.结论 ERCC1 mRNA的表达水平可以作为以铂类为基础药物化疗的NSCLC患者预后的独立预测因素,ERCC1 mRNA低表达的NSCLC患者经铂类化疗后,总生存时间明显延长,为制定NSCLC个体化的化疗方案提供了重要信息.     

非小细胞肺癌组织ERCC1和RRM1基因表达及其对预后影响探讨

目的比较非小细胞肺癌（non-small cell lung cancer,NSCLC）患者原发灶与转移淋巴结中切除修复交叉互补基因1（excision repair cross-complementation group1,ERCC1）和核糖核苷酸还原酶M1亚基（ribonucleotidereductase subunit M1,RRM1）表达情况,探讨ERCC1和RRM1表达状态与NSCLC患者治疗效果及预后的关系,为NSCLC患者的合理治疗提供依据。方法选取山东省肿瘤医院外科六病区2008-08-01-2012-08-31行手术切除且术后病理确诊为NSCLCⅡA-ⅢA期的患者106例,所有患者术前均未接受任何针对肿瘤的治疗措施,均釆用免疫组化技术测定组织中ERCC1和RRM1表达状况,术后均采用长春瑞滨联合顺铂（NP方案）进行辅助化疗,并进行临床随访。结果 NSCLC癌组织及转移淋巴结中RRM1阴性表达患者中位无疾病进展时间分别为23.1和24.7个月,显著优于RRM1阳性表达患者的16.4和19.1个月,两组比较差异有统计学意义,P值分别为0.007和0.026;而ERCC1阴性表达患者中位无疾病生存期为17.5个月,与ERCC1阳性表达患者的19.4个月比较,差异无统计学意义,P=0.59。原发灶和淋巴结转移灶中ERCC1和RRM1的表达水平差异无统计学意义,P〉0.05。不同性别、分期及病理类型的NSCLC患者ERCC1及RRM1表达水平差异均无统计学意义,P〉0.05。结论 NSCLC患者癌组织及转移淋巴结中,RRM1基因表达情况可作为NP方案术后辅助化疗预后的重要指标。