某地区结直肠癌患者KRAS基因及BRAF基因突变状态的检测分析

目的 检测某地区结直肠癌患者KRAS基因和BRAF基因的突变状态及其与年龄、性别的关系.方法 由某地区30例结直肠癌患者石蜡组织中提取DNA,通过直接测序法及荧光定量PCR法检测KRAS基因及BRAF基因突变状态.结果 KRAS基因突变率为43.3%,共发现6种突变类型,主要位于12、13密码子,其中以c.38G>A突变率最高(38.5%).单因素及多因素分析均提示KRAS突变与年龄或性别无相关性.BRAF基因突变率为0.结论 某地区结直肠癌患者KRAS基因突变率高,靶向治疗前进行突变状态检测具有重要临床价值.     

原发性肝癌中KRAS及BRAF基因突变检测的临床意义

目的探讨分析KRAS及BRAF基因突变检测对原发性肝癌的意义。方法回顾性分析2017年1月-2019年10月我院收治的158例原发性肝癌患者,根据病理临床证实是否存在肝外转移,分为转移组86例、非转移组72例,采集所有患者的血液,检测KRAS、BRAF基因以及突变情况,分析两组KRAS、BRAF基因的突变率以及突变的情况。结果转移组KRAS基因突变率明显比非转移组高(P<0.05),两组BRAF基因均未出现突变(P>0.05)。结论有肝外转移的患者KRAS存在突变,且突变率较高,而转移和非转移均未出现BRAF基因突变,因此可根据KRAS基因突变来判断原发性肝癌是否存在转移,其对原发性肝癌是否存在转移中具有临床价值。     

结直肠黏液腺癌临床病理与MMR蛋白表达及KRAS、BRAF基因突变的相关性分析

目的 分析结直肠黏液腺癌临床病理因素与MMR蛋白表达及KRAS、BRAF基因突变的相关性。方法 回顾性分析2014-01—2022-01山东省东阿县人民医院手术切除的33例黏液腺癌和307例非黏液腺癌患者的临床病理资料，采用免疫组化法和扩增阻滞突变系统(ARMS)对340例结直肠癌标本石蜡组织的MMR蛋白表达及KRAS、BRAF基因突变状态检测，并对其进行相关性分析。结果 黏液腺癌占结直肠癌的9.7%(33/340),与肿瘤大小、病变部位、浸润深度、错配修复蛋白表达及BRAF基因突变等临床病理因素有相关性(P<0.05)。结论 黏液腺癌是结直肠癌的少见组织学亚型，分析临床病例特征与错配修复蛋白联合KRAS、BRAF突变检测可为患者预后评估及个体化治疗提供依据。     

HRM法检测结直肠癌患者循环DNA中KRAS和EGFR基因突变

摘要:目的：探讨高分辨率熔解曲线(HRM) 法检测结直肠癌患者循环DNA中KRAS和EGFR基因突变的可行性。 方法：用HRM法检测70例结直肠癌患者KRAS基因2号和3号外显子以及EGFR基因19和21号外显子的突变情况,用直接测序法验证结果的准确性。 结果：70 例结直肠癌患者血液标本经HRM法、直接测序法分别检测到18例(25.7%)和17例(24.3%)KRAS基因2号外显子突变;均检测到3例EGFR基因19号外显子突变(4.3%);两法均未检测到3号外显子和21号外显子突变。HRM检测与直接测序法结果符合率为99.6% (279/280)。 结论：HRM法检测结直肠癌患者血样本KRAS和EGFR基因突变,具有准确度高、操作简单、检测成本低等优点,适合在临床推广应用。     

肺癌患者EGFR及KRAS基因突变检测方法的建立和临床应用

目的 本研究拟在评估二代测序(next generation sequencing,NGS)技术在肺癌临床分子诊断中应用的可行性. 方法 本研究选取108例肺癌石蜡包埋样本(formalin fixed paraffin embedded,FFPE),同时进行一代测序(sanger sequencing)和NGS检测样本中EGFR和KRAS基因的突变情况. 结果 在108例肺癌样本中,一代测序检出突变在二代测序中均检出,另外NGS还检测出目标区域外其他突变. 结论 NGS的检测准确性可达到100％.NGS可以应用于肺癌临床分子诊断,同时NGS能给临床提供更详细的基因突变信息,为分子靶向治疗提供依据,更具有市场潜力及应用前景.     

结肠直肠癌KRAS、NRAS、BRAF基因突变及其与临床病理特征、p53蛋白表达间相关性研究

目的：筛查肠直肠癌患者的KRAS、NRAS和BRAF基因亚型突变状态, 并分析其与患者临床病理特征及p53蛋白表达情况间的相关性。方法：收集110例手术切除的结肠直肠癌病理资料及组织学样本, 总结临床病理特征;采用突变阻滞扩增系统法检测KRAS、NRAS基因2、3、4号外显子第12、13、61、117、146密码子及BRAF基因15号外显子第600密码子的突变情况;采用免疫组织化学（免疫组化）法检测p53蛋白的表达情况。结果：KRAS基因突变率为45.4%（50/110）, 其中49例患者检测到2号外显子第12和13密码子突变, 1例检测到4号外显子第117和146密码子突变;NRAS基因突变率为6.3%（7/110）, 其中3例检测到2号外显子第12和13密码子突变, 3例检测到3号外显子第61密码子突变, 1例检测到4号外显子第146密码子突变, 且与KRAS基因存在共突变;BRAF基因15号外显子第600密码子突变率为2.7%（3/110）;p53蛋白阳性表达率为57.3%（63/110）, 其中27例患者存在KRAS基因突变, 7例存在NRAS基因突变, 2例存在BRAF基因突变。KRAS基因突变率与患者的淋巴结转移显著相关（P<0.05）;而NRAS基因突变与其组织分型及p53蛋白表达显著相关（P<0.05）;BRAF基因突变率与右半结肠癌显著相关（P<0.05）。结论：KRAS与结肠直肠癌发生、发展的分子机制关系密切;BRAF基因突变仅见于右半结肠, 提示右半结肠癌发生机制可能有其特殊性;p53蛋白表达与NRAS基因突变之间是否存在相互作用关系值得进一步研究。     

结直肠癌患者KRAS、NRAS、BRAF及PIK3CA基因突变与其临床病理特征关系的研究

目的 探讨结直肠癌(colorectal cancer, CRC)患者KRAS、NRAS、BRAF、PIK3CA基因突变情况及其与临床病理特征的关系。方法 收集2012—2016年复旦大学附属上海市第五人民医院手术切除的91例原发性CRC患者肿瘤手术切除标本进行回顾性分析,采用扩增阻碍突变系统(ARMS)进行实时荧光定量PCR检测KRAS、NRAS、BRAF、PIK3CA基因突变情况,应用统计学分析其与临床病理学特征的关系。结果 91例CRC患者中,KRAS/NRAS/BRAF/PIK3CA总突变率为53.8%(49/91),其中KRAS突变率为40.7%(37/91),NRAS突变率为4.4%(4/91),BRAF突变率为3.3%(3/91),PIK3CA突变率为2.2%(2/91),KRAS/PIK3CA双突变率为2.2%(2/91),KRAS/NRAS双突变率为1.1%(1/91)。BRAF基因在低分化癌患者中突变率显著高于中-高分化癌患者(P<0.05)。PIK3CA基因突变与肿瘤原发部位有关(P<0.05)。有淋巴结转移患者的KRNS/NRAS/BRAF/PIK3...     

结直肠癌微卫星不稳定状态与KRAS、NRAS、BRAF基因突变和临床病理特征及预后的关系

目的 探讨结直肠癌(CRC)的微卫星不稳定(MSI)状态与KRAS、NRAS、BRAF基因突变、临床病理特征及预后的相关性。方法 选取2015年3月至2020年12月于汕头市中心医院和中山大学肿瘤防治中心收治的经病理确诊为CRC的861例患者作为研究对象,检测其4种错配修复(MMR)蛋白表达、MSI状态,KRAS、NRAS、NRAS基因突变情况。分析MMR蛋白表达、KRAS、NRAS和BRAF基因突变与CRC患者临床病理特征的相关性,MMR蛋白表达与KRAS、NRAS和BRAF基因突变的相关性,并采用COX回归模型探讨CRC患者2年内死亡的独立影响因素。结果 CRC中的MMR蛋白表达总缺失率为12.08%,KRAS基因突变率为42.28%,NRAS基因突变率为2.21%,BRAF基因突变率为6.50%。不同年龄、肿瘤部位、组织学类型、肿瘤最大径、分化程度、TNM分期、淋巴结转移状态、CEA水平的患者MMR蛋白表达比较,差异有统计学意义(P<0.05)。不同性别、肿瘤部位、组织学类型、分化程度、TNM分期、淋巴结转移状态、CA199水平的患者KRAS基因突变比较,差异有统计学意义(...     

高分辨率熔解曲线分析法检测非小细胞肺癌KRAS和BRAF基因突变

目的探讨高分辨率熔解曲线分析(HRM)法检测非小细胞肺癌(NSCLC)中KRAS和BRAF基因突变用于临床检测的可行性。方法用HRM法检测64例NSCLC患者KRAS基因第2外显子和BRAF基因第15外显子的突变情况,用直接测序法对结果进行验证。结果 HRM法检测结果表明有9例NSCLC患者发生KRAS基因突变(14.06%)、4例发生BRAF基因突变(6.25%),直接测序法证实两法的结果完全一致;共检测出4种KRAS基因突变类型,G12C(GGT>TGT)的突变率最高(44.4%),BRAF基因突变型均为V600E。结论用HRM法检测临床样本KRAS和BRAF基因突变,具有操作简便、结果准确、成本低的优点,适用于临床检测。     

非小细胞肺癌EGFR和KRAS基因突变和临床病理特征分析

目的 探讨非小细胞肺癌(NSCLC)中表皮生长因子受体(EGFR)和鼠类肉瘤病毒癌基因(KRAS)突变状态及其与临床病理特征的关系.方法 回顾性收集2019年1月至2021年6月首都医科大学大兴教学医院183例病理确诊NSCLC患者临床病理资料,采用扩增阻滞突变系统(ARMS)法检测肿瘤组织中EGFR和KRAS基因的突...     

KRAS/BRAF mutations in brain arteriovenous malformations: A systematic review and meta-analysis

IntroductionSomatic KRAS mutations have been identified in the majority of brain arteriovenous malformations (AVM) specimens. The aim of our study was to evaluate the prevalence of Kirsten rat sarcoma (KRAS)/murine sarcoma viral oncogene homolog B1 (BRAF) mutations in brain AVM.MethodsA systematic literature review was performed in November 2019. We reviewed MEDLINE/PubMed, Cochrane Library, and ClinicalTrials.gov for citation or ongoing trials from January 2010 to March 2020.Results6 studies were identified as meeting the inclusion criteria of this review. The total frequency of KRAS mutations in 1726 patients with AVM was 55%. The prevalence of BRAF mutation was 7.5%. The prevalence of AVMs with grade 2 was the most (39%). Frontal and parietal lobes were the commonest sites of AVMs (21%). the most prevalent presentation of patients with AVM was hemorrhage (62%).ConclusionOur findings support a high prevalence of somatic activating mutations in KRAS and less commonly, BRAF in the overwhelming majority of brain AVMs. Practically and importantly, this pathway homogeneity in CNS arteriovenous malformations also supports the development of targeted therapies with RAS/RAF pathway inhibitors. However, more studies are needed to confirm this hypothesis.     

A new generation of companion diagnostics: cobas BRAF,KRAS and EGFR mutation detection tests

The cobas^® (Roche) portfolio of companion diagnostics in oncology currently has three assays CE-marked for in vitro diagnostics. Two of these (EGFR and BRAF) are also US FDA-approved. These assays detect clinically relevant mutations that are correlated with response (BRAF, EGFR) or lack of response (KRAS) to targeted therapies such as selective mutant BRAF inhibitors in malignant melanoma, tyrosine kinases inhibitor in non-small cell lung cancer and anti-EGFR monoclonal antibodies in colorectal cancer, respectively. All these assays are run on a single platform using DNA extracted from a single 5 µm section of a formalin-fixed paraffin-embedded tissue block. The assays provide an ‘end-to-end’ solution from extraction of DNA to automated analysis and report on the cobas z 480. The cobas tests have shown robust and reproducible performance, with high sensitivity and specificity and low limit of detection, making them suitable as companion diagnostics for clinical use.     

EGFR mutations and EGFR tyrosine kinase inhibition in non-small cell lung cancer

OBJECTIVES: To discuss selected molecular targets and new clinical variables that can serve as both predictive and prognostic markers for outcome in patients with non-small cell lung cancer (NSCLC) treated with EGFR-TKIs. DATA SOURCES: Research and journal articles. CONCLUSION: In the near future, treatment for NSCLC will rely ever increasingly on molecular targets rather than empirically chosen cytotoxic chemotherapy for some patients. This will improve outcomes for patients with NSCLC. IMPLICATIONS FOR NURSING PRACTICE: An understanding of the molecular targets and clinical variables that are predictive and prognostic of outcome in NSCLC will help nurses better care for these patients.     

Mutation patterns and prognostic analysis of BRAF/KRAS/PIK3CA in colorectal cancer

Background and objectiveAberrant gene expression and abnormal signaling pathways often occur in patients with colorectal cancer, in which mutations in B‐Raf Proto‐Oncogene (BRAF), KRAS Proto‐Oncogene (KRAS), and Phosphatidylinositol‐4,5‐Bisphosphate 3‐Kinase Catalytic Subunit Alpha (PIK3CA) are quite common. In this study, the relationship between BRAF, KRAS, and PIK3CA mutations and clinicopathologic features and prognosis of colorectal cancer patients was investigated.MethodsOne hundred and fifty patients with colorectal cancer admitted to Affiliated people''s Hospital (Fujian Provincial People''s Hospital), Fujian University of Traditional Chinese Medicine were collected and grouped according to the mutation patterns of BRAF, KRAS, and PIK3CA. The association between BRAF, KRAS, and PIK3CA mutations and pathological factors (age, sex, etc.) was analyzed using the Chi‐square test. Subsequently, survival analysis was performed to screen the impact factors of overall survival time by Kaplan–Meier (K‐M) curve, and Cox regression model was established for the selected factors.ResultsBRAF, KRAS, and PIK3CA mutations were not associated with age, sex, and alcoholism. K–M curve and log‐rank test results demonstrated that among the factors included in this study, overall survival rate of colorectal cancer patients was only associated with mutation factors. The prognosis of KRAS+/PIK3CA−/BRAF−mutant and KRAS−/PIK3CA−/BRAF+mutant patients was better than that of KRAS+/PIK3CA+/BRAF−mutant patients.ConclusionThe mutant patterns of BRAF, KRAS, and PIK3CA were not related to the general and clinicopathological features of patients. The mutant pattern could be used as an independent prognostic factor for colorectal cancer.