ERα、EGFR 及 CyclinD1在上皮性卵巢癌中的表达及意义

目的：探讨雌激素受体α（ ERα）、表皮生长因子受体（ EGFR）及细胞周期蛋白D1（ CyclinD1）在上皮性卵巢癌中的表达及临床意义。方法采用免疫组化SP法观察ERα、EGFR及CyclinD1在上皮性卵巢癌组织58份（浆液性卵巢癌组织38份、非浆液性卵巢癌组织20份）、正常卵巢组织18份、卵巢良性肿瘤组织20份中的表达情况。结果 ERα、EGFR及CyclinD1在上皮性卵巢癌组织中的阳性表达率分别为74．13％、68．97％、51．72％,显著高于正常卵巢组织（33．33％、16．67％、0）及卵巢良性肿瘤组织（30．0％、25．0％、10．0％）（P均＜0．05）;但ERα、EGFR及CyclinD1在卵巢良性肿瘤、正常卵巢组织中的阳性表达率比较P均＞0．05。 ERα、EGFR及CyclinD1在上皮性卵巢癌中的表达与临床分期及细胞分化程度有关（P均＜0．05）。结论 ERα、EGFR及CyclinD1在上皮性卵巢癌中过表达,并在一定程度上反映其恶性程度及预后状况。     

卵巢上皮性癌组织中EGFR、STAT3 mRNA的表达及意义

目的观察卵巢上皮性癌（卵巢癌）组织中表皮生长因子受体（EGFR）与信号转导和转录激活因子3（STAT3）mRNA的表达,并探讨其意义。方法用RT-PCR技术检测13例正常卵巢、11例卵巢良性肿瘤及43例卵巢癌组织中的EGFR mRNA和STAT3 mRNA。结果卵巢上皮性癌组织中EGFR、STAT3 mRNA阳性表达率显著高于正常卵巢及卵巢良性肿瘤组织（P均〈0.05）。卵巢上皮性癌组织中EGFR、STAT3 mRNA表达均与卵巢癌病理分期有关,与患者年龄、肿瘤分化程度、组织学类型及有无淋巴结转移等临床病理参数无关（P均〉0.05）;卵巢癌组织中EGFR和STAT3 mRNA表达有显著相关性（r=0.42,P〈0.01）。结论卵巢癌组织中EGFR、STAT3表达上调。二者在卵巢癌的发生发展中起重要作用。     

卵巢癌组织中P-糖蛋白、肺耐药蛋白的表达变化及意义

目的探讨原发性上皮性卵巢癌（POEC）组织中P-糖蛋自（P—gP）及肺耐药蛋白（LRP）的表达及意义。方法采用免疫组化法检测56例POEC、15例卵巢良性肿瘤、12例正常卵巢组织中的P—gP、HIP,并分析其与POEC患者临床病理参数的关系。结果P-gp、LRP在POEC组织中的阳性表达率显著高于卵巢良性肿瘤和正常卵巢组织（P均〈0．01）;LRP的表达与POEC患者腹腔积液有关（P〈0．01）;术前化疗的POEC患者P-gp、LRP阳性表达率高于初治者（P〈0．01）,P-gp、LRP表达阳性的POEC对化疗反应差。结论P-gp、LRP在POEC组织中的阳性表达率高于卵巢良性肿瘤和正常卵巢组织;化疗可使POEC组织P-gp、LRP的阳性表达率升高;P-gp及LRP的表达可预测POEC对铂类药物化疗的疗效。     

卵巢癌中γ-synuclein、MMP-9、MVD的表达及意义

采用免疫组化sP法对50例卵巢癌组织、15例卵巢良性肿瘤组织和5例正常卵巢组织中的γ-synuclein（SNCG）、基质金属蛋白酶（MMP）及微血管密度（MVD）进行检测。认为SNCG、MMP-9及MVD在卵巢癌中阳性表达率增高，有淋巴系统转移者和晚期癌，阳性表达率增高（P均〈0．05）。MVD与MMP-9表达呈正相关。SNCG、MMP-9与MVD将有可能作为卵巢癌早期诊断指标之一。     

整合素α5β1和FAK在上皮卵巢肿瘤组织中的表达及意义

目的研究整合素α5β1和黏着斑激酶（FAK）在卵巢上皮肿瘤组织中的表达及其与临床病理特征的关系。方法应用免疫组化方法检测49例卵巢上皮癌组织（观察组）和15例卵巢上皮良性肿瘤组织（对照组）中整合素α5β1和FAK的表达情况。结果观察组整合素α5β1和FAK的表达明显高于对照组（P〈0．05）;整合素α5β1阳性表达率与卵巢上皮癌组织的分化程度以及临床分期有关（P均〈0．05）,与患者年龄、肿瘤大小及组织学分型无关（P〉0．05）;FAK阳性表达率与卵巢上皮癌组织的临床分期有关（P〈0．05）,与患者年龄、分化程度、肿瘤大小及组织学分型无关（P〉0．05）;整合素α5β1、FAK在卵巢上皮癌组织中的表达呈正相关（r=0．490,P〈0．01）。结论整合素α5β1、FAK在卵巢上皮癌组织中的表达具有相关性,与卵巢上皮癌的发生、侵袭和转移密切相关。     

卵巢癌组织中PCNA、Ki-67的表达变化及意义

目的观察卵巢癌组织中增殖细胞核抗原（PCNA）、增殖细胞相关核抗原（Ki-67）的表达变化,并探讨其临床意义。方法采用免疫组化SP法检测40例卵巢癌、20例卵巢良性肿瘤组织中的PCNA、Ki-67。结果 PC-NA、Ki-67在卵巢癌组织中的阳性表达率明显高于卵巢良性肿瘤（P均〈0.05）;PCNA、Ki-67的表达与卵巢癌FIGO分期、组织分级、淋巴结转移有关（P均〈0.05）;卵巢癌组织中PCNA与Ki-67的表达呈正相关（r=0.929,P〈0.05）。结论卵巢癌组织中PCNA、Ki-67的表达增加,二者可作为评估卵巢癌临床分期、组织分级及淋巴结转移的参考指标。     

卵巢癌组织中P185、ER和PR的表达及意义

采用间接免疫荧光流式细胞半定量分析技术检测卵巢癌42例、卵巢良性肿瘤8例及正常卵巢组织10例中ERBB-2、雌激素受体（ER）、孕激素受体（PR）的表达以及三者的相关性。P185、ER、PR在卵巢恶性肿瘤组织中的表达量明显高于良性卵巢组织及正常卵巢组织（P〈0．05）,而良性肿瘤组织与正常组织中表达量无明显差异（P〉0．05）,并且其表达量与组织学类型和临床分期无关,三者具有相关性。认为卵巢癌不论其病理分级及组织类型,P185、ER、PR均呈高表达,可作为卵巢癌诊断、病情评估、预后判断的指标。     

AnnexinⅡ、Caspase-3蛋白在卵巢癌组织中的表达及意义

目的进一步探讨卵巢癌的发生机制。方法选择同期手术切除的组织标本,卵巢癌52份（观察组）、卵巢良性上皮性肿瘤18份（对照组）,采用免疫组化SP法检测其AnnexinⅡ、凋亡蛋白酶（Caspase）-3蛋白表达情况,采用Spearman相关分析法分析两指标间及其与卵巢癌临床病理特征的关系。结果观察组与对照组AnnexinⅡ蛋白阳性表达率分别为80．77％（42／52）、44．45％（8／18）,Caspase-3蛋白阳性表达率分别为48．08％（25／52）、94．44％（17／18）,两两比较P均〈0．05;AnnexinⅡ蛋白表达与卵巢癌组织学类型、组织学分级无关,与临床分期和淋巴结转移有关（P〈0．05）,Caspase-3蛋白表达与卵巢癌组织类型、病理学分级、淋巴结转移无关,与卵巢癌临床分期有显著相关性（P〈0．05）;Annexin11和Caspase-3蛋白在卵巢癌组织中的表达呈负相关（r=-0．409,P〈0．05）。结论AnnexinⅡ、Caspase,3蛋白异常表达在卵巢癌发生、发展中起重要作用,且两者存在协同性。     

凋亡抑制基因Livin在卵巢癌组织中的表达

应用逆转录-聚合酶链反应（RT—PCR）和免疫印迹（Western blot）法检测Livin在20例卵巢癌组织中及10例正常卵巢组织的表达情况。结果发现,20例卵巢癌组织中Livin基因高表达,正常的卵巢癌组织Livin均无明显表达。20例卵巢癌组织中Livin蛋白阳性表达共有12例（60％）,10例正常卵巢组织中Livin蛋白阳性表达仅有1例（10％）,明显低于卵巢癌组织（P〈0．05）。认为Livin的表达与卵巢癌的发生、发展有关,检测卵巢癌组织中Livin表达可能对判断卵巢癌预后有一定意义。     

膀胱移行细胞癌组织中Hec1、CyclinD1的表达变化及意义

目的 观察癌症高表达蛋白（Hec1）和细胞周期蛋白D1（CyclinD1）在膀胱移行细胞癌（TCC）组织中的表达变化,并探讨其临床意义。方法采用免疫组化sP法检测15例正常膀胱黏膜、85例TCC组织及20例癌旁组织中的Hec1和CyclinD1。结果TCC组织中Hec1、CyclinD1阳性表达率均明显高于正常膀胱组织及癌旁组织（P均〈0．05）。CyclinD1的表达与TCC病理分级、临床分期有关（P均〈0．05）,Hec1的表达与TCC临床分期有关（P〈0．05）。TCC组织中Hec1和CyclinD1的表达呈正相关（r=0．265,P〈0．05）。结论Hec1和CyclinD1存TCC中讨嘉扶．可作为TCC病弹分级和临床分期的判断指标．     

卵巢肿瘤组织中EGFR、CK20表达及意义

目的探讨细胞角质蛋白20（CK20）和表皮生长因子受体（EGFR）mRNA在卵巢上皮性肿瘤发生中的作用。方法应用RT-PCR法检测12份正常卵巢（对照组）、18份卵巢良性肿瘤（良性组）、33份卵巢恶性肿瘤（恶性组）和14份胃肠道转移性卵巢肿瘤（转移组）组织中CK20、EGFR mRNA的表达。结果恶性组中CK20、EGFR mRNA阳性率显著高于对照组及良性组（P〈0.05）,与转移组比较无明显差异;恶性组CK20 mRNA表达与手术病理分期、淋巴结转移有关（P〈0.05）,EGFR mRNA表达与手术病理分期有关（P〈0.05）。结论CK20、EG-FR高表达可促进卵巢恶性肿瘤的发生、转移;其检测对判断预后有一定价值。     

Prognostic and predictive biomarkers for epidermal growth factor receptor-targeted therapy in colorectal cancer: Beyond KRAS mutations

The advent of the epidermal growth factor receptor (EGFR)-targeted monoclonal antibodies (mAbs), cetuximab and panitumumab has expanded the range of treatment options for metastatic colorectal cancer (CRC). Despite these agents have paved the way to individualized therapy, our understanding why some patients respond to treatment whereas others do not remain poor. The realization that detection of positive EGFR expression by IHC does not reliably predict clinical outcome of EGFR-targeted treatment has led to an intense search for alternative predictive biomarkers. Data derived from multiple phase III trials have indicated that KRAS mutations can be considered a highly specific negative biomarker of benefit to anti-EGFR mAbs. Oncologists are now facing emerging issues in the treatment of metastatic CRC, including the identification of additional genetic determinants of primary resistance to EGFR-targeted therapy for further improving selection of patients, the explanation of rare cases of patients carrying KRAS-mutated tumours who have been reported to respond to cetuximab and panitumumab and the discovery of mechanisms of secondary resistance to EGFR-targeted therapy. Current data suggest that, together with KRAS mutations, the evaluation of EGFR gene copy number (GCN), BRAF, NRAS, PIK3CA mutations or loss of PTEN expression could also be useful for selecting patients with reduced chance to benefit from anti-EGFR mAbs.This review aims to provide an updated of the most recent data on predictive and prognostic biomarkers within the EGFR pathway, the challenges this emerging field presents and the future role of these molecular markers in CRC treatment.     

Identification of EGFR and KRAS mutations in Chinese patients with esophageal squamous cell carcinoma

The prognosis of esophageal squamous cell carcinoma (ESCC) is poor. It is urgent to improve this situation. Epidermal growth factor receptor (EGFR)‐targeted therapy possesses a promising clinical efficacy. Mutations of EGFR and V‐Ki‐ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) have been identified in esophageal carcinoma, but corresponding Chinese data are limited. So we investigated the mutation status of EGFR and KRAS in Chinese patients with ESCC, and explored their correlations with clinicopathological features. Formalin‐fixed paraffin‐embedded surgically resected tumor samples were obtained from 50 randomly selected Chinese patients with ESCC. EGFR mutations in exons 18–21 were detected by Scorpions amplification refractory mutation system technology. KRAS mutations in codons 12, 13 were detected by direct sequencing of polymerase chain reaction products. The correlations between clinicopathological features and the mutation status of EGFR and KRAS were analyzed using the Statistical Package for the Social Sciences. In the present study, EGFR mutations were found in 7 (14%) out of 50 patients, including G719X missense mutation (n= 1), in‐frame deletion (n= 2), and L858R missense mutation (n= 5). Six (12%) out of 50 patients had KRAS mutations in codon 12. Concurrent EGFR and KRAS mutations were detected in one sample. The presences of EGFR and KRAS mutations were not associated with gender, age, smoking history, cell differentiation, or cancer stage. In conclusion, the incidence of EGFR mutations in Chinese patients with ESCC was higher than that of previous reports, and the incidence of KRAS mutations was not low. EGFR and KRAS mutations were mainly located in exons 19 and 21 and codon 12, respectively. Unlike in NSCLC, concurrent EGFR and KRAS mutations existed.     

Superoxide dismutase 1 (SOD1) is a target for a small molecule identified in a screen for inhibitors of the growth of lung adenocarcinoma cell lines

We previously described four small molecules that reduced the growth of lung adenocarcinoma cell lines with either epidermal growth factor receptor (EGFR) or KRAS mutations in a high-throughout chemical screen. By combining affinity proteomics and gene expression analysis, we now propose superoxide dismutase 1 (SOD1) as the most likely target of one of these small molecules, referred to as lung cancer screen 1 (LCS-1). siRNAs against SOD1 slowed the growth of LCS-1 sensitive cell lines; conversely, expression of a SOD1 cDNA increased proliferation of H358 cells and reduced sensitivity of these cells to LCS-1. In addition, SOD1 enzymatic activity was inhibited in vitro by LCS-1 and two closely related analogs. These results suggest that SOD1 is an LCS-1-binding protein that may act in concert with mutant proteins, such as EGFR and KRAS, to promote cell growth, providing a therapeutic target for compounds like LCS-1.     

KRAS p.G13D mutations are associated with sensitivity to anti-EGFR antibody treatment in colorectal cancer cell lines

Purpose

Targeted therapies using the anti-EGFR antibodies panitumumab (Pmab) or cetuximab (Cmab) are currently restricted to patients with metastatic colorectal adenocarcinoma whose tumours do not show a mutation in KRAS. However, recent retrospective studies indicated that patients with tumours mutated in codon 13 of KRAS may benefit from treatment with Cmab in contrast to patients with tumours mutated in KRAS codon 12.

Methods

To study the functional impact of the subtype of KRAS mutations on the efficiency of EGFR-targeted therapies, we correlated the KRAS mutation status of 15 colorectal carcinoma cell lines with the in vitro sensitivity of these cells to Cmab/Pmab. Mutations in the potential predictive biomarkers BRAF and PIK3CA as well as protein expression of EGFR and PTEN were also determined.

Results

Four out of seven KRAS-mutated cell lines were characterised by the p.G13D mutation. Treatment of these cells using Cmab/Pmab induced a significant growth inhibition in contrast to cell lines showing a KRAS mutation at codon 12 or 61. Out of the eight KRAS wild-type cell lines, five were insensitive to Cmab/Pmab. These cell lines were characterised either by BRAF mutation or by absence of EGFR or PTEN protein expression.

Conclusions

Since KRAS p.G13D-mutated tumour cells may respond to EGFR-targeted therapy, we suggest including subtype analysis of KRAS mutations in prospective clinical trials. In KRAS wild-type tumour cells, BRAF mutations and loss of EGFR or PTEN expression may lead to resistance to EGFR-targeted therapy and should be considered as additional negative predictive biomarkers.     

KRAS和表皮生长因子受体突变基因与原发性非小细胞肺癌的相关性

目的 研究KRAS和表皮生长因子受体(EGFR)基因在原发性非小细胞肺癌(NSCLC)中的突变状态以及其与靶向治疗效果的相关性。方法 收集郑州大学第一附属医院收治的150例经病理证实的NSCLC患者原发肿瘤及受累的淋巴结标本,进行KRAS和EGFR突变基因序列分析。150例中12例患者纵隔淋巴结转移灶经DNA序列分析发现EGFR基因突变后给予吉非替尼进行术前辅助治疗。结果150对（分别为原发灶和受累淋巴结）标本中,2例原发肿瘤和10例淋巴结转移灶中检测到KRAS突变基因,35例原发肿瘤和44例淋巴结转移灶中检测到EGFR突变基因。KRAS和EGFR基因在原发灶和转移灶中不一致的比率分别为6.7％ (10/150)和8.7％ (13/150)。1例原发肿瘤无EGFR突变的患者接受吉非替尼治疗后效果不佳。结论 NSCLC患者原发灶和转移灶中KRAS和EGFR基因突变状态不一致,这对于应用酪氨酸激酶抑制剂靶向治疗NSCLC具有重要的参考意义。     

p21-activated kinase 4 regulates ovarian cancer cell proliferation, migration, and invasion and contributes to poor prognosis in patients

Ovarian cancer is a lethal gynecological malignancy, and to improve survival, it is important to identify novel prognostic and therapeutic targets. In this study, we present a role for p21-activated kinase 4 (Pak4) in ovarian cancer progression. We show a significant association between increased expression of Pak4 and its activated form, phosphorylated (p)-Pak4 Ser(474), with metastasis of ovarian cancers, shorter overall and disease-free survival, advanced stage and high-grade cancers, serous/clear cell histological subtypes, and reduced chemosensitivity. Pak4 overexpression was also observed in ovarian cancer cell lines. Pak4 and p-Pak4 expression were detected both in the nucleus and cytoplasm of ovarian cancer cells, in vitro as well as in vivo. Stable knockdown of Pak4 in ovarian cancer cell lines led to reduced cell migration, invasion, and proliferation, along with reduced c-Src, ERK1/2, and epidermal growth factor receptor (EGFR) activation and decreased matrix metalloproteinase 2 (MMP2) expression. Conversely, Pak4 overexpression promoted ovarian cancer cell migration and invasion in a c-Src, MEK-1, MMP2, and kinase-dependent manner, and induced cell proliferation through the Pak4/c-Src/EGFR pathway that controls cyclin D1 and CDC25A expression. Stable knockdown of Pak4 also impeded tumor growth and dissemination in nude mice. This report reveals the association between Pak4 and important clinicopathologic parameters, suggesting Pak4 to be a significant prognostic marker and potential therapeutic molecular target in ovarian cancer. The implied possible cross-talk between Pak4 and EGFR suggests the potential of dual targeting of EGFR and Pak4 as a unique therapeutic approach for cancer therapy.