The expression of hepatocyte growth factor (HGF) and c-Met in uterine serous carcinoma

Objective

The hepatocyte growth factor (HGF) receptor c-Met plays an important role in tumor dissemination by activating mitogenic signaling pathways. The goal of this study was to investigate immunohistochemical (IHC) staining patterns of HGF and c-Met in endometrioid endometrial cancer (EC) and uterine serous cancer (USC) and to correlate staining with patient outcomes.

Methods

A tissue microarray was created using tissue from patients with atrophic endometrium (AE), grade 1 EC, grade 3 EC, and USC. Immunohistochemistry was used to detect c-Met, phosphorylated c-Met (p-c-Met), and HGF expression. Differences between IHC staining intensity were calculated using t-tests. Correlations between staining and clinicopathologic variables were determined by Chi-square testing for categorical variables and t-tests for continuous variables. Kaplan-Meier curves were constructed to analyze survival in USC.

Results

Patients with cancer had more total c-Met and HGF expression than those with AE (p = 0.037, p < 0.001 respectively), but no difference in p-c-Met staining. Total c-Met and HGF staining was significantly different between groups (p = 0.042, p < 0.001 respectively). This difference was accounted for by greater c-MET expression in USC compared to AE (p = 0.027). Depth of invasion, stage, and lymph node status were not significantly related to staining intensity. Patients with strong c-Met and HGF staining had decreased overall survival compared to patients with weaker staining.

Conclusions

HGF and c-Met may play a role in the progression of endometrial cancer and should be studied further as prognostic and therapeutic tools.     

Met in lung cancer

Receptor tyrosine kinases play important roles in the biology of many tumor cell types. In approximately 10% of non-small cell lung cancer (NSCLC) patients mutational activation of the epidermal growth factor receptor (EGFR) results in tumor cells that are exquisitely addicted to signaling by this receptor.¹ Thus expression of mutant active EGFR but in general not wild-type EGFR predisposes NSCLC cells to inhibitors of EGFR/ErbB2. Use of EGFR inhibitory agents such as gefitinib for this subset of NSCLC patients causes tumor regression and disease stabilization for 12–18 mo, after which tumor cells become resistant to the drug.² Initial studies identified a second mutation within the EGFR, which results in the resistance of the tyrosine kinase to gefitinib, as a major cause of reduced tumor control.³ This has resulted in the development of newer EGFR inhibitors, e.g., afatinib, which inhibited double mutant EGFR.⁴ In a subset of these patients, however, resistance to gefitinib was not associated with EGFR mutations.⁵ Clearly, other mechanisms of gefitinib resistance must be at play.     

c—Met、PTEN在甲状腺癌中的表达及其临床意义

目的研究c—Met、PTEN在甲状腺癌中的表达和临床价值。方法采用免疫组织化学SP法检测c—Met、PTEN在甲状腺癌、甲状腺腺瘤及正常甲状腺组织中的表达,并对其在不同病理类型、临床分期的甲状腺癌中的标记结果进行对照和相关性分析。结果c—Met、PTEN在72例甲状腺癌中的阳性率分别为63．9％和62．5％,与正常甲状腺、甲状腺腺瘤分别相比,差异有统计学意义（P〈0．05）。其在不同病理类型、临床分期、病理分级和区域淋巴结转移的甲状腺癌病例中的表达结果相比,差异有统计学意义（P〈0．05）,而与肿瘤大小、瘤体有无侵出包膜、患者性别及年龄无关（P〉0．05）。c—Met与PTEN的表达均呈负相关（r=-0．340,P〈0．01）。结论c—Met、PTEN在甲状腺癌的衍变过程中可能扮演重要角色,二者联合标记可能作为评估甲状腺癌恶性度及预后的有效参考,并可能为其靶向治疗提供思路。     

CDl51、c-Met及整合素仪α3、α6在胰腺导管腺癌中的表达及其与预后的关系

目的探讨CD蚓、c-Met和整合素仪3、拍蛋白在胰腺导管腺癌中的表达及其与预后的关系。方法用免疫组化法检测71例胰腺导管腺癌及10例正常胰腺组织中CD151、c-Met和整合素仪3、嘶蛋白的表达,分析它们与临床病理特征及患者预后的关系。结果CD151、c-Met和整合素仪3、面在71例胰腺导管腺癌组织中的表达阳性率分别为81．69％（58／71）、69．01％（49／71）、69．01％（49／71）和84．51％（60／71）,而正常胰腺组织均未表达。CD151和c-Met的表达与肿瘤的TNM分期、淋巴结转移显著相关（P值均〈0．05）。CD151,的表达与c-Met及整合素α3、α6的表达呈正相关（r=0．583,P=0．000;r=0．457;P=0．000;r=0．671;P=0．000）。单因素分析显示,CD151、c-Met、整合素α3和α6的表达与预后有关（P值均〈0．05）。多因素分析表明,CD151、c-Met是患者术后生存时间的独立预后因子。结论CD151、c-Met及整合素α3、α6在胰腺癌的发展、转移及预后发挥重要作用,CD151、c-Met可考虑作为临床评价胰腺癌生物学行为及评估预后的指标。     

HGF、c-Met和VEGF-C在宫颈癌中的表达

 目的检测宫颈癌组织中HGF及其受体原癌基因c-Met和VEGF-C的分子表达水平及三者间的关系。方法 采用实时荧光定量RT-PCR(Real Time fluorescence Quantitative PCR,FQ RT-PCR)技术相对定量检测43 例宫颈浸润癌,30例CIN Ⅲ和27例正常宫颈组织标本中HGF mRNA、c-Met mRNA和VEGF-C mRNA的表达水平 。结果HGF mRNA、c-Met mRNA和VEGF-C mRNA在正常宫颈、CIN Ⅲ和宫颈浸润癌组织中的表达水平依次升 高,差异有统计学意义(P<0.05)。宫颈癌组织中三者表达水平与肿瘤临床分期、肿瘤直径大小、浸润深度 、淋巴结转移有关(P均<0.05)。 HGF mRNA与c-Met mRNA和VEGF-C mRNA表达相关(P<0.01)。结论HGF、c- Met和VEGF-C在宫颈癌发生、发展、转移中发挥重要作用,它们参与了宫颈癌新生血管和淋巴管的形成, 对宫颈癌的诊断、治疗及预后有指导意义。     

Prognostic significance of overexpression of c-Met oncoprotein in cholangiocarcinoma

Background:

Cholangiocarcinoma (CC) is a highly malignant carcinoma. We attempted to clarify the prognostic significance of c-Met overexpression and its association with clinicopathological factors in patients with CC.

Patients and methods:

One hundred and eleven patients with intrahepatic CC (IHCC) and 136 patients with extrahepatic CC (EHCC) who had undergone curative surgery were divided immunohistologically into c-Met^high and c-Met^low groups. Clinicopathological factors and outcomes were compared between the groups. c-Met and epidermal growth factor receptor (EGFR) expression was also examined in 10 CC cell lines.

Results:

The positivity of c-Met was 45.0% in IHCC and 68.4% in EHCC; c-Met^high expression was demonstrated in 11.7% of IHCC and 16.2% of EHCC. c-Met^high expression was significantly correlated with the 5-year survival rate for CC overall (P=0.0046) and for IHCC (P=0.0013), histopathological classification in EHCC, and for EGFR overexpression in both IHCC and EHCC. Coexpression and coactivation of c-Met and EGFR were also observed in CC cell lines. Multivariate analysis revealed that c-Met^high expression was an independent predictor of poor overall and disease-free survival in patients with IHCC.

Conclusions:

c-Met overexpression is associated with EGFR expression and is a poor prognostic factor in CC.     

Ezrin和c-Met在胃癌组织中的表达及意义

目的研究Ezrin和c-Met在胃癌组织中的表达及其临床意义。方法应用免疫组织化学法检测45例胃癌、25例胃腺瘤、20例正常胃黏膜组织中Ezrin和c-Met的表达水平,分析Ezrin和c-Met的表达与各临床病理因素的关系。结果胃癌组织中Ezrin和c-Met的表达水平明显高于胃腺瘤组织及正常胃黏膜组织(P<0.05)。Ezrin和c-Met的表达水平与肿瘤的浸润深度、分化程度及有无淋巴结转移/远处转移有关(P<0.05),与性别、年龄及肿瘤大小无关(P>0.05),两者在胃癌组织中的表达呈正相关关系(r=0.603,P<0.001)。结论 Ezrin和c-Met与胃癌的发生、侵袭、转移密切相关,二者在胃癌发生、发展过程中可能具有协同作用,联合检测Ezrin和c-Met的表达水平可能为胃癌预后判断提供依据。     

pSilencer2.0-c-Met-siRNA基因在裸鼠体内对人喉癌Hep-2细胞生长的影响

目的探讨pSilencer2.0-c-Met-siRNA重组质粒对人喉癌Hep-2细胞裸鼠移植瘤生长的影响。方法采用裸鼠喉癌皮下荷瘤模型模拟c-Met-siRNA基因治疗实验,观察c-Met-siRNA重组质粒的抗瘤疗效,Western-blot法检测重组质粒对c-Met基因及MMP-2、MMP-9、VEGF基因表达的影响。结果肿瘤接种后第35 d,重组质粒组肿瘤体积为13 827 mm^3,与对照组相比差异有统计学意义（P〈0.01）;重组质粒组瘤体内c-Met、MMP-2、MMP-9、VEGF基因表达率比对照组明显降低（P〈0.05）。结论pSilencer2.0-c-Met-siRNA重组质粒能明显抑制人喉癌Hep-2细胞移植裸鼠成瘤后的生长,siRNA表达质粒介导的基因治疗有希望成为喉癌靶向性c-Met治疗的新策略。     

重组免疫毒素抗c-Met/PE38KDEL免疫毒素的构建?表达及纯化

目的:构建绿脓杆菌外毒素PE38KDEL融合全人源抗c-Met单链抗体原核表达载体,并对其产物进行变性、复性和纯化,检测其对肝癌细胞系的毒性作用.方法:以从人源天然Fab抗体库中筛选出的AM2-26克隆作为模板扩增出人源c-Met单链基因,采用基因工程原理,将扩增出的c-Met单链基因和PE38KDEL基因克隆入pBAD/GⅢA表达载体中,重组克隆载体经酶切及DNA序列测定证实两片段连接正确;转化大肠杆菌Top10F',左旋阿拉伯糖诱导表达,采用镍离子螯合层析法纯化变性的包涵体样品,并用连续梯度透析的方法对纯化后的包涵体进行复性;采用流式细胞术鉴定复性产物与靶细胞的结合活性,MTT法检测c-Met/PE38KDEL免疫毒素对肝癌细胞的体外杀伤活性.结果:成功构建了免疫毒素表达载体pBAD/GⅢA/c-Met/PE38KDEL;诱导产物主要以包涵体形式存在;复性后的免疫毒素c-Met/PE38KDEL具有与靶细胞特异性结合的活性并且对SMMC7721细胞系具有较明显的杀伤作用.结论:全人源抗c-Met单链抗体与PE38KDEL重组免疫毒素的成功表达纯化及对人肝癌细胞系的试验结果,为进一步肝癌的导向治疗提供依据.