胃肠道间质瘤中c-kit和血小板衍生生长因子受体α基因的突变及其表达研究

目的探讨c-kit和血小板衍生生长因子受体α（PDGFR—α）基因突变及其蛋白表达在胃肠道问质瘤（GIST）发病中的作用及与其临床病理、预后的关系。方法应用免疫组织化学EnvisionTM二步法检测了119例GIST中CD117和PDGFR-α蛋白的表达情况。用PCR扩增和基因测序的方法，检测50例GIST c—kit基因第9、11、13、17号外显子以及PDGFR—α基因第12、18号外显子的突变情况。结果本组119例GIST中CD-117的阳性表达率为87．4％，PDGFR-α的阳性表达率为65．5％。50例GIST中c—kit基因突变率为42％。突变均位于11号外显子近膜区的5’端第556～560密码子之间，即所谓的突变“热点”。50例GIST中PDGFR-α基因突变率为20％，最常见的突变为第18号外显子D842V点突变，2例为第12号外显子突变。c-kit基因突变主要见于CD117阳性的GIST，与GIST生物学侵袭行为无明显关系；而PDGFR-α基因突变主要见于CD117阴性的GIST，具高度侵袭危险性。未发现1例GIST有c—kit和PDGFR-α基因同时突变。结论PDGFR-α作为一种GIST特异而敏感的标记物，对一些CD117表达阴性的GIST的诊断及鉴别诊断具有重要的临床意义。作为GIST发病的两种分子机制，c—kit和PDGFR—α基因突变是相互排斥，不共存的。     

胃肠道间质瘤c—kit及PDGFRA基因突变及其临床意义

目的探讨胃肠道间质瘤（GIST）中c-kit及PDGFRA基因突变的分子生物学特点及与临床参数间的关系。方法对141例GIST进行基因检测,用PCR扩增和基因测序的方法检测肿瘤c-kit基因第9、11、13和17外显子及PDGFRA基因第12和18外显子的序列,分析基因突变与GIST临床参数之间的关系。结果在141例GIST中共检测到c—kit基因突变108例（76．6％）,其中11号外显子突变99例（70．2％,99／141）;9号外显子突变8例（5．7％,8／141）;13号外显子突变1例（0．7％,1／141）;未检测到17号外显子突变病例。97．0％为杂合性突变,3．0％为纯合性突变,11外显子突变方式以缺失突变最常见65．7％（65／99）,其次为点突变（24．2％）和插入突变（串联重复）（10．1％）;突变位点多集中在5’端的经典热区,其次为3’端的框内串联重复。共检测到4例PDGFRA基因的突变．占无c—kit突变病例的12．1％（4／33）,占CD117阴性GIST的40％（4／10）,均为外显子18的突变。基因突变的发生率在GIST不同原发部位间差异有统计学意义（χ^2=7．229,P=0．027;χ^2=7．000,P=0．03）,而与患者年龄、性别、肿瘤大小、核分裂、恶性潜能分级等均差异无统计学意义。结论GIST中存在c-kit及PDGFRA基因的突变;基因突变的发生率在不同原发部位间有差异。     

磷酸化血小板源生长因子受体α和C—kit在胃肠道和胃肠道外间质瘤的表达及其临床意义

目的探讨磷酸化血小板源生长因子受体α（P—PDGFR—α）在胃肠道间质瘤（GIST）和胃肠道外GIST（EGIST）的表达及临床意义，为进一步提高GIST和EGIST的病理诊断、病理分型及临床治疗提供依据。方法用免疫组织化学方法检测28例CD117阳性和13例CD117阴性的GIST和EGIST间质瘤组织中P—PDGFR-α的表达，并且用PCR直接测序的方法检测41例GISTC—kiI基因外显子9、11、13、17和P—PDGFR-α外显子12、18突变。结果P—PDGFR-α在CD117阴性的GIST表达（69．2％）显著高于在CD117阳性的GIST表达（7．1％）（P〈0．05）；P—PDGFR-α在上皮型GIST的表达（27．3％）和混合型GIST的表达（63．3％）均显著高于在梭型细胞型GIST的表达（9％）（P〈0．05）；CD117在梭型细胞型GIST的表达（53．6％）明显高于在上皮型GIST（7．1％）和混合型GIST（39．3％）的表达（P〈0．05）。在28例CD117阳性GIST中，19例有c—kit基因的突变，其中15例在外显子11有突变，4例在外显子13有突变，13例CD117阴性GIST中无C—kit基因突变：11例PDGFR-α阳性的GIST中4例有PDGFR-α基因的突变．均发生在外显子18。结论P—PDGFR-α为CD117阴性GIST的病理诊断、病理分型和临床治疗进一步提供了可靠的依据。PDGFR-α基因突变后引起产物蛋白的磷酸化可能是CD117阴性的GIST发生的重要分子基础和生物学行为。     

C—kit与血小板源性生长因子受体基因突变特征与胃肠间质瘤患者预后的关系

目的探讨C-kit与血小板源性生长因子受体（PDGFRA）基因突变特征及其与胃肠间质瘤（GIST）患者预后的关系。方法收集2000年6月至2009年1月山西省肿瘤医院收治的99例GIST患者的I临床病理、基因检测及随访资料。应用Kaplan．Meier法计算生存率并进行单因素分析,cox比例风险模型进行多因素分析。结果99例患者的5年无瘤生存率（DFS）为61．5％;5年总体生存率（OS）为67．4％。其中,77例c-kit外显子11、4例c．kit外显子9和2例PDGFRA外显子18突变患者5年DFS分别为64．3％、14．3％和100％。5年OS分别为70．8％、50．0％和100％。在c-kit外显子11突变的类型中。26例点突变、44例删失突变和7例复制突变患者5年DFS分别为87．1％、44．9％和80．0％,5年OS分别为88．1％、57．0％和100％。各因素之间5年DFS和OS差异均有统计学意义（P〈0．05）。多因素分析示,基因突变并不是预后的独立影响因素（P=0．492）。结论经手术治疗而未服用伊马替尼的GIST患者．基因突变型预后优于野生型。但基因突变并非其预后的独立影响因素。     

基因突变与胃肠道间质瘤研究进展

摘要：c kit及血小板源性生长因子受体α（platelet derived growth factor receptor alpha,PDGFRA）的功能增强性突变是导致胃肠道间质瘤（GIST）的主要原因。c kit基因及PDGFRA基因的突变位点及突变方式对肿瘤的临床特征有明显影响。笔者就基因突变与GIST的关系进行综述。     

胃肠道间质瘤c-kit基因突变的研究进展

目的探讨胃肠道间质瘤（gastrointestinal stromal tumor,GIST）c-kit基因突变特征与临床病理、分子靶向治疗及预后的相关性。方法采用文献复习的方法,对研究胃肠道间质瘤分子遗传学机理的相关文献加以综述。结果 c-kit基因突变可能是GIST发生的早期事件,是GIST的普遍现象,不能作为GIST预后判断的指标。然而,c-kit突变位点和方式是否影响GIST的生物学行为,能否作为评估预后的指标仍存在争议。结论基因突变检测对从分子水平上研究疾病、判断预后以及指导用药具有重要意义,需要不断地深入研究。     

c-kit基因突变对胃肠道间质瘤预后的影响

目的　探讨c kit基因突变与胃肠道间质瘤 (GIST)临床病理指标和预后的关系 ,为GIST恶性变判断、预后评估提供客观指标。方法　对 82例GIST患者运用聚合酶链反应 单链构象多态性分析、DNA测序的方法筛选c kit基因突变病例 ,采用统计学方法回顾性分析c kit癌基因突变与GIST临床病理特征、生物学行为、复发和病死率的关系。结果　c kit基因突变阳性组与c kit基因突变阴性组间肿瘤大小 (χ2 =16 795 ,P <0 0 1)、增殖细胞核抗原指数 (χ2 =17 0 99,P <0 0 1)、有丝分裂计数 (χ2 =11 2 33,P <0 0 1)、坏死 (χ2 =4 4 0 4 ,P <0 0 5 )、浸润 (χ2 =2 2 391,P <0 0 1)、复发 (χ2 =16 333,P <0 0 1)、转移 (χ2 =12 96 5 ,P <0 0 1)、死亡 (χ2 =11 6 4 6 ,P <0 0 1)差异有显著意义 ,而年龄 (χ2 =0 0 0 0 2 ,P >0 0 5 )、性别 (χ2 =0 14 1,P >0 0 5 )、肿瘤的部位 (χ2 =7 5 2 4 ,P >0 0 5 )、细胞类型 (χ2 =0 82 7,P >0 0 5 )、囊性变 (χ2 =1 6 0 8,P >0 0 5 )、出血 (χ2 =0 6 33,P >0 0 5 )、c kit蛋白表达 (χ2 =0 0 0 0 ,P >0 0 5 )差异无显著意义。结论　c kit基因突变的检测可作为GIST患者预后评估的有意义指标     

腹腔镜手术治疗胃肠道间质瘤

胃肠道间质瘤（gastrointestinal stromal turnors,GIST）是一种较为少见的胃肠道非上皮源性的间充质肿瘤。由于其组织来源复杂，以往常被诊断为平滑肌瘤，平滑肌肉瘤或神经鞘瘤等。94％的GIST由于c—kit基因突变，导致酪氨酸激酶受体kit过度表达。因此，目前主要依靠其分子特点以及免疫组化表现对GIST作出病理诊断。     

胃肠道间质瘤c-kit和PDGFRA基因突变检测及临床诊断意义

目的 观察c-kit基因和PDGFRA基因在胃肠道间质瘤(GIST)中的突变.方法 分析23例GIST患者的临床病理资料,检测23例GIST c-kit基因9、11、13和17号外显子突变以及PDGFRA基因12和18号外显子突变.结果 23例GIST中kit基因突变率为70.0%,其中11、9、13号外显子突变分别为56.5%、8.7%、4.3%,未检测到17号外显子的突变.PDGFRA基因突变率为13.0%,其中18、12号外显子突变分别为8.7%、4.3%.结论 GIST病例大多数存在c-kit基因和PDGFRA基因的突变,CD117阳性GIST主要表现为c-kit突变,以11号外显子突变最为常见,PDGFRA基因突变主要见于CD117阴性GIST.

Abstract：

Objective To explore the starus of activating mutations of c-kit and PDGFRA in gastrointestinal stromal tumor (GIST) of Chinese patients.Methods The clinic and pathological data of a series of patients with gastrointestinal stromal tumors in 2008-2009 yrs were analyzed retrospectively.Exon 9,11,13,17 of c-kit and exon 12,18 of PDGFRA were analyzed by direct sequencing.Results c-kit mutations were detected in 70.0% of patients as follows :56.5% in exon 11 ,8.7% in exon 9,4.3% in exon 13 and none in exon 17.PDGFRA mutations were present in 13.0% of CD1 17-negative GIST,8.7% in exon 18,4.3% in exon 12.Conclusion The vast majority of GIST in this study harbored c-kit and PDGFRA mutation.Most CD117 expressing GIST show c-kit mutations that are preferentially in exon 11.PDGFRA genic mutations are more likely seen in kit-negative GISTs.     

胃肠道间质瘤的临床诊断和分期

胃肠道间质瘤（gastrointestinal stromal tumors，GIST）是源于胃肠道间质细胞的肿瘤，多由c-kit基因突变引起；细胞形态呈梭形或上皮样；表达kit蛋白，免疫组化特异性CD117，阳性；手术切除后易复发和转移，传统放疗和化疗效果差。虽然是消化道的少见肿瘤，但是随着诊断标准的明确，GIST病例数有日益增多的趋势；且作为肿瘤分子靶向治疗的热点，有关GIST的研究正受到国内外肿瘤外科的广泛关注。     

原癌基因c-kit在胃肠道间质瘤中的表达及其临床意义

目的　通过检测胃肠道间质瘤 (gastrointestinalstromaltumor ,GIST)中原癌基因c kit蛋白表达 ,探讨c kit在GIST中的诊断及预后意义。　方法　应用免疫组织化学技术EnVision微波二步法检测了 12 2例GIST(良性 5 4例 ,潜在恶性 13例 ,恶性 5 5例 )中的c kit蛋白的表达情况 ,并与平滑肌瘤、神经鞘瘤进行对照研究。不着色或仅细胞核着色者为阴性 ,凡肿瘤细胞浆内或 (和 )细胞膜上有棕黄色颗粒者为阳性。　结果　GIST中c kit的阳性表达率为 97% (118/ 12 2 ) ,阳性信号定位准确。其中 ,良性组、潜在恶性组及恶性组c kit的阳性率分别为 98% (5 3/ 5 4 )、93% (12 / 13)、96 % (5 3/ 5 5 )。与良性组相比 ,转移或复发病例c kit蛋白着色较淡 ,但 3组之间c kit的表达水平差异无显著意义 (χ2 =1 16 7,P >0 0 5 )。平滑肌瘤和神经鞘瘤c kit阴性表达。　结论　c kit作为一种胃肠道间质瘤特异而敏感的标记物 ,对GIST的诊断及鉴别诊断有重要的意义 ,但其不能作为GIST分化程度的指标     

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目的探讨胃肠间质瘤病人的临床病理特征及c-kit/血小板源性生长受体α(PDGFRA)基因突变频率和突变类型。方法收集2002年1月至2011年10月于北京大学肿瘤医院就诊并经病理组织学确诊的660例胃肠间质瘤病人的临床病理资料及肿瘤组织标本,运用直接测序方法检测组织标本中c-ki(t外显子9、11、13和17)、PDGFRA基因(外显子12和18)的突变状态,统计学分析基因型与临床病理特征的相关性。结果 660例胃肠间质瘤病人中,男389例(58.9%),女271例(41.1%)。中位年龄56(15～82)岁。原发部位以胃(37%)和小肠(35%)为主,转移部位以肝脏及腹腔最常见。酪氨酸激酶受体(CD117)阳性率95.0%,DOG1阳性率88.1%。360例病人接受基因突变检测,其中c-kit基因外显子11突变241例(66.9%),外显子9突变43例(11.9%),外显子13突变6例(1.7%),外显子17突变4例(1.1%),PDGFRA基因外显子12突变1例(0.3%),外显子18突变7例(1.9%),野生型病人58例(16.2%);c-kit外显子9突变主要见于小肠,而PDGFRA12/18突变主要见于胃;基因突变频率与肿瘤大小与和分裂像未见相关性。结论胃肠间质瘤最常发生在胃和小肠;c-kit基因外显子11为最常见的基因突变类型;基因突变类型与原发部位相关。     

KISS1蛋白在胃肠道间质瘤中的表达及其预后预测意义

目的 检测胃肠道间质瘤(GIST)组织中KISS1蛋白的表达,分析其与GIST患者预后的关系.方法 连续收集1987年1月至2008年9月北京大学人民医院诊断治疗的GIST病例137例,以及非GIST的间叶源性肿瘤73例,采用免疫组织化学染色方法检测KISS1蛋白的表达情况,分析KISS1蛋白的表达与GIST临床病理指标及预后的关系.结果 KISS1蛋白在本组GIST组织中的表达率为40.9%,高于非GIST间叶源性肿瘤组织(P＜0.05);GIST组织中KISS1蛋白表达率与肿瘤长径、病变范围、细胞丰富程度、有无假包膜、Fletcher危险程度分级及术后转移相关(P＜0.05);KISS1蛋白阴性GIST患者的预后好于KISS1蛋白阳性患者(P＜0.05).结论 KISS1在蛋白水平的表达与GIST临床病理指标相关,与GIST患者术后转移的发生及预后相关,有可能作为新的判断GIST预后的指标.

Abstract：

Objective To evaluate KISS1 expression, and its significance in the prognosis of GIST patients. Methods In this study, 137 GIST cases and 73 non-GIST sarcoma cases were evaluated for clinicopathological characteristics and immunohistochemistry for KISS1 antibodies. Result The expression rate of KISS1 was 40.9% (56/137) in GISTs,which was significantly correlated with tumor size, disease extent, cellularity, presence of pseudocapaule, Fletcher's risk stratification and metastatic status after resection (P＜0.05). Patients with positive KISS1 expression had significantly worse disease free survival and disease specific survival (P ＜ 0.05 ). Conclusions KISS1 expression was associated with some clinicopathological characteristics as well as malignant behaviors in patients with GISTs. KISS1 might be a predictor in prognosis for GIST patients.     

Mutations in exon 11 of the c-kit gene in a myogenic tumor and a neurogenic tumor as well as in gastrointestinal stromal tumors. Utility of c-kit mutation as a prognostic biomarker for gastrointestinal mesenchymal tumor

BACKGROUND/AIMS: Gain-of-function mutations in exons 9, 11 and 13 of the c-kit gene in gastrointestinal stromal tumors (GISTs) have been identified, and it has been reported that the prognosis is worse for patients with mutation-positive GISTs than for those with mutation-negative GISTs. We studied c-kit mutations in gastrointestinal mesenchymal tumors. By chance, the c-kit mutation in exon 11 was found in myogenic and neurogenic tumors as well as in GISTs. Furthermore, we studied the clinical prognostic utility of these mutations. METHODS: Ten gastrointestinal mesenchymal tumors were stained with HE and immunohistochemically analyzed with alpha-smooth muscle actin, S-100 protein, CD34 and c-kit. In these tumors, as well as in 11 cases of leiomyomas, PCR-amplified DNA from the juxtamembrane (JM) domain of exon 11, the extracellular domain of exon 9 and the tyrosine kinase domain 1 of exon 13 showed a high frequency of c-kit mutation and was sequenced. RESULTS: Although c-kit mutations have previously been reported only in GISTs, we found c-kit mutations in the JM domain of exon 11 in one myogenic and one neurogenic tumor as well as in two GISTs. No c-kit mutation was seen in the 11 cases of leiomyomas. In addition, all four cases with c-kit mutation in exon 11 suffered a relapse sooner than the other cases without c-kit mutations. CONCLUSION: Clinically, the prognosis was worse for the patients with mutation-positive gastrointestinal mesenchymal tumors than for those with mutation-negative tumors. We therefore conclude that the gain-of-function mutation in exon 11 of the c-kit gene is an important prognostic factor for gastrointestinal mesenchymal tumors, including myogenic and neurogenic tumors as well as GISTs.     

Predicting the Antitumor Effects of STI571 by Analysis of c-kit Gene Mutations in Gastrointestinal Stromal Tumors of the Stomach: Report of a Case

We report a case of a large gastrointestinal stromal tumor (GIST), greater than 5 cm in diameter, in the stomach. Microscopically, high levels of mitosis were observed, indicative of a high-grade malignancy. We analyzed the c-kit gene mutations by a replication competent retrovirus assay and DNA sequencing, which revealed a c-kit mutation in exon 11. Liver metastases were detected 7 months after surgery. Patients with an exon 11 mutation of the c-kit gene are reported to have a high response to STI571 (imatinib mesylate, Glivec). Accordingly, a 1-month course of STI571 treatment clearly changed the characterization of the metastatic tumors radiographically. Thus, it may be important to analyze c-kit gene mutations in patients presenting with GISTs to predict the effectiveness of STI571 in suppressing GISTs, especially tumors thought to have malignant potential.     

c-kit and PDGFRA mutations in extragastrointestinal stromal tumor (gastrointestinal stromal tumor of the soft tissue)

Extragastrointestinal stromal tumor (EGIST) is a unique tumor that occurs outside the gastrointestinal tract. EGIST shows a c-kit expression and histologic appearance similar to those of gastrointestinal stromal tumor (GIST). Most GISTs have gain-of-functional mutation of the c-kit gene, and some have mutation of the platelet-derived growth factor receptor-alpha (PDGFRA) gene. However, the frequency of mutation of those genes in EGISTs remains unclear. We examined the clinicopathologic features, prognostic factors, and c-kit and PDGFRA mutation in 39 cases of EGIST. Tumors with high mitotic counts (>or=5/50 high power fields) or a high Ki-67 labeling index (>or=10%) were significantly correlated with worse prognoses. The c-kit mutation was found in the juxtamembrane domain (exon 11) and the extracellular domain (exon 9) in 12 of 29 cases (41.4%) and 2 of 29 cases (6.9%), respectively. The PDGFRA gene mutation was found at the juxtamembrane domain (exon 12) and the tyrosine kinase domain (exon 18) in one case each. The pattern of kit and PDGFRA mutation in EGIST was essentially similar to that in GIST. Our results suggest that the c-kit and PDGFRA mutations play an important role in the tumorigenesis of EGIST. High mitotic counts and a high Ki-67 labeling index may be useful for predicting the aggressive biologic behavior in EGIST. Furthermore, STI-571, targeting c-kit and PDGFR tyrosine kinase, seems to be a possible therapeutic strategy for EGISTs, especially advanced cases.