食管鳞癌的蛋白质组学

大部分食管鳞癌（esophageal squamous cell carcinoma, ESCC）确诊时已发展至中晚期,临床治疗效果差,是导致我国华北地区ESCC死亡率居高不下的主要原因之一.因此,亟需筛查ESCC特异性、敏感性的生物标志物,以期用于ESCC早期诊断、个体化分子靶向治疗和预后评价. 与相对稳定、携带遗传信息的基因组不同,蛋白质组具有时空变化特性,由此构成生命活动复杂性的物质基础.在病理情况下,蛋白质组能够精确反映患病组织器官的功能状态,因此为疾病的监测提供了窗口.本文总结了ESCC蛋白质组研究现状及差异表达的蛋白质谱,并探讨了ESCC候选分子标志物的潜在临床应用价值.     

术后放化疗在局部晚期食管鳞状细胞癌中的临床应用研究

目的:研究局部晚期食管鳞状细胞癌术后放化疗对患者生存率的影响研究.方法:选择我院2009年1月～2012年1月经根治术后病理学检查确诊的局部晚期食管鳞状细胞癌患者150例,按知情同意原则分成单纯放疗组(48例)、单纯化疗组(54例)、放化疗组(48例)三组,单纯放疗组于术后6周内用15MV-X线照射,总剂量50Gy/25f,治疗30天.单纯化疗组于术后4周内开始用DDP和5-Fu联合方案进行序贯性化疗,放化疗组的剂量同单纯放疗组和单纯化疗组.结果:三组患者治疗有效率分别为72.91％、51.85％、91.67％,差别有统计学意义(P＜0.05),三组患者的三年生存率组间差别有统计学意义(P＜0.05),放化疗组患者治疗有效率及三年生存率最高.结论:局部晚期食管鳞状细胞癌术后应用放化疗作为辅助治疗手段疗效较单纯化疗和单纯放疗好.     

食管鳞癌组织中的神经纤维分布

目的:探讨食管鳞癌组织中神经纤维的分布情况.方法:应用免疫组化ABC法,探查手术切除的食管鳞癌组织里S100及GAP-43阳性神经纤维的分布情况,并分析其与患者临床病理参数的关系.结果:相对于正常组织,食管鳞癌组织中存在相当数量的S100及GAP-43阳性神经纤维(束)不规则地分布于肿瘤细胞之间,且S100阳性纤维密度大于GAP-43阳性纤维密度;统计分析显示肿瘤组织中纤维密度与患者的肿瘤大小、淋巴结转移相关.结论:食管鳞癌组织中确实存在神经纤维分布,并对肿瘤发展起一定作用.     

食管鳞癌组织中的神经纤维分布

目的：探讨食管鳞癌组织中神经纤维的分布情况。方法：应用免疫组化ABC法,探查手术切除的食管鳞癌组织里S100及GAP-43阳性神经纤维的分布情况,并分析其与患者临床病理参数的关系。结果：相对于正常组织,食管鳞癌组织中存在相当数量的S100及GAP-43阳性神经纤维（束）不规则地分布于肿瘤细胞之间,且S100阳性纤维密度大于GAP-43阳性纤维密度;统计分析显示肿瘤组织中纤维密度与患者的肿瘤大小、淋巴结转移相关。结论：食管鳞癌组织中确实存在神经纤维分布,并对肿瘤发展起一定作用。     

Sparse Representation-Based Patient-Specific Diagnosis and Treatment for Esophageal Squamous Cell Carcinoma

Precision medicine and personalized treatment have attracted attention in recent years. However, most genetic medicines mainly target one genetic site, while complex diseases like esophageal squamous cell carcinoma (ESCC) usually present heterogeneity that involves variations of many genetic markers. Here, we seek an approach to leverage genetic data and ESCC knowledge data to forward personalized diagnosis and treatment for ESCC. First, 851 ESCC-related gene markers and their druggability were studied through a comprehensive literature analysis. Then, a sparse representation-based variable selection (SRVS) was employed for patient-specific genetic marker selection using gene expression datasets. Results showed that the SRVS method could identify a unique gene vector for each patient group, leading to significantly higher classification accuracies compared to randomly selected genes (100, 97.17, 100, 100%; permutation p values: 0.0032, 0.0008, 0.0004, and 0.0008). The SRVS also outperformed an ANOVA-based gene selection method in terms of the classification ratio. The patient-specific gene markers are targets of ESCC effective drugs, providing specific guidance for medicine selection. Our results suggest the effectiveness of integrating previous database utilizing SRVS in assisting personalized medicine selection and treatment for ESCC.     

Prognostic Nomogram for Thoracic Esophageal Squamous Cell Carcinoma after Radical Esophagectomy

Nomogram has demonstrated its capability in individualized estimates of survival in diverse cancers. Here we retrospectively investigated 1195 patients with esophageal squamous-cell carcinoma (ESCC) who underwent radical esophagectomy at Zhejiang Cancer Hospital in Hangzhou, China. We randomly assigned two-thirds of the patients to a training cohort (n = 797) and one-third to a validation cohort (n = 398). Cox proportional hazards regression analyses were performed using the training cohort, and a nomogram was developed for predicting 3-year and 5-year overall survival rates. Multivariate analysis identified tumor length, surgical approach, number of examined lymph node, number of positive lymph node, extent of positive lymph node, grade, and depth of invasion as independent risk factors for survival. The discriminative ability of the nomogram was externally determined using the validation cohort, showing that the nomogram exhibited a sufficient level of discrimination according to the C-index (0.715, 95% CI 0.671–0.759). The C-index of the nomogram was significantly higher than that of the sixth edition (0.664, P-value<0.0001) and the seventh edition (0.696, P-value<0.0003) of the TNM classification. This study developed the first nomogram for ESCC, which can be applied in daily clinical practice for individualized survival prediction.     

食管鳞癌与腺癌差异基因表达的对比分析

目的:研究食管鳞癌(esophageal squamous cell carcinoma,ESCC)与食管腺癌(esophageal adenocarcinoma,EAC)的基因差异表达,探讨ESCC与EAC发生发展的基因学基础。方法:选取8例ESCC和8例EAC组织抽提mRNA,应用cDNA芯片技术通过芯片杂交、生物信息学处理,找出两者间差异表达基因。结果:采用BioStarH-40芯片发现差异表达基因541条,差异表达基因占13.8%,其中表达增强309条(显著增强73条),表达降低232条(显著降低61条)。结论:ESCC与EAC基因表达比较,差异有统计学意义,这些差异可能在两类肿瘤不同的生物学行为中起重要作用。     

食管鳞癌与腺癌差异基因表达的对比分析

目的：研究食管鳞癌（esophageal squamous cell carcinoma,ESCC）与食管腺癌（esophageal adenocarcinoma,EAC）的基因差异表达,探讨ESCC与EAC发生发展的基因学基础。方法：选取8例ESCC和8例EAC组织抽提mRNA,应用cDNA芯片技术通过芯片杂交、生物信息学处理,找出两者间差异表达基因。结果：采用BioStarH-40芯片发现差异表达基因541条,差异表达基因占13．8％,其中表达增强309条（显著增强73条）,表达降低232条（显著降低61条）。结论：ESCC与EAC基因表达比较,差异有统计学意义,这些差异可能在两类肿瘤不同的生物学行为中起重要作用。     

Preclinical Validation of Talaporfin Sodium-Mediated Photodynamic Therapy for Esophageal Squamous Cell Carcinoma

Photodynamic therapy (PDT) kills cancer cells via a photochemical reaction mediated by an oncotropic photosensitizer. Herein, we performed an experimental preclinical study to validate the anti-tumour effect of talaporfin sodium-mediated PDT (t-PDT) for esophageal squamous cell carcinoma (ESCC) cells. We used human ESCC cells derived from various differentiation grades or resistant to 5-fluorouracil (5-FU). The cytotoxic effect of t-PDT was determined by evaluating cell viability, apoptosis and generation of reactive oxygen species (ROS) and DNA double-strand breaks. Furthermore, the anti-tumour effect of t-PDT was assessed using an anchorage-independent cell-growth assay and xenograft transplantation models. t-PDT induced potent cytotoxicity in ESCC cells independent of their differentiation grade or 5-FU resistance. Moreover, t-PDT induced robust apoptosis, as indicated by cell shrinkage, perinuclear vacuolization, nuclear fragmentation and induction of annexin V-positive cells. This apoptotic response was accompanied by concurrent activation of ROS, and induction of DNA double-strand breakage. Importantly, t-PDT suppressed efficiently anchorage-independent cell growth as well as ESCC-xenografted tumor formation. In aggregate, t-PDT showed anti-tumor potential for ESCC cells with various histological grades or chemoresistance, providing a novel translational rationale of t-PDT for the treatment of ESCC.     

Tenascin-C,a Prognostic Determinant of Esophageal Squamous Cell Carcinoma

Background

Tenascin-C, an adhesion modulatory extracellular matrix molecule, is highly expressed in numerous human malignancies; thus, it may contribute to carcinogenesis and tumor progression. We explored the clinicopathological significance of Tenascin-C as a prognostic determinant of esophageal squamous cell carcinoma (ESCC).

Methods

In ESCC patient tissues and cell lines, the presence of isoforms were examined using western blotting. We then investigated Tenascin-C immunohistochemical expression in 136 ESCC tissue samples. The clinical relevance of Tenascin-C expression and the correlation between Tenascin-C expression and expression of other factors related to cancer-associated fibroblasts (CAFs) were also determined.

Results

Both 250 and 350 kDa sized isoforms of Tenascin-C were expressed only in esophageal cancer tissue not in normal tissue. Furthermore, both isoforms were also identified in all of four CAFs derived from esophageal cancer tissues. Tenascin-C expression was remarkably higher in ESCC than in adjacent non-tumor esophageal epithelium (p < 0.001). Tenascin-C expression in ESCC stromal fibroblasts was associated with patient’s age, tumor (pT) stage, lymph node metastasis, clinical stage, and cancer recurrence. Tenascin-C expression in cancer cells was correlated with an increase in tumor-associated macrophage (TAM) population, cancer recurrence, and hypoxia inducible factor1α (HIF1α) expression. Moreover, Tenascin-C overexpression in cancer cells and stromal fibroblasts was an independent poor prognostic factor for overall survival (OS) and disease-free survival (DFS). In the Cox proportional hazard regression model, Tenascin-C overexpression in cancer cells and stromal fibroblasts was a significant independent hazard factor for OS and DFS in ESCC patients in both univariate and multivariate analyses. Furthermore, Tenascin-C expression in stromal fibroblasts of the ESCC patients was positively correlated with platelet-derived growth factor α (PDGFRα), PDGFRβ, and smooth muscle actin (SMA) expression. The 5-year OS and DFS rates were remarkably lower in patients with positive expressions of both Tenascin-C and PDGFRα (p < 0.001), Tenascin-C and PDGFRβ (p < 0.001), Tenascin-C and SMA (p < 0.001), Tenascin-C and fibroblast activation protein (FAP) (p < 0.001), and Tenascin-C and fibroblast-stimulating protein-1 (FSP1) (p < 0.001) in ESCC stromal fibroblasts than in patients with negative expressions of both Tenascin-C and one of the abovementioned CAF markers.

Conclusion

Our results show that Tenascin-C is a reliable and significant prognostic factor in ESCC. Tenascin-C may thus be a potent ESCC therapeutic target.     

A Panel of Overexpressed Proteins for Prognosis in Esophageal Squamous Cell Carcinoma

Esophageal squamous cell carcinoma (ESCC) is a common cancer with poor prognosis. In order to identify useful biomarkers for accurately classifying prognostic risks for ESCC patients, we examined the expression of six proteins by immunohistochemistry (IHC) in 590 paraffin-embedded ESCC samples. The candidate proteins include p53, EGFR, c-KIT, TIMP1 and PI3K-p110α reported to be altered in ESCC tissues as well as another important component of PI3K, PI3K-p85α. Of the six proteins tested, p53, EGFR, c-KIT, TIMP1 and PI3K-p85α were detected with high expression in 43.0%, 36.6%, 55.9%, 70.7% and 57.1% of tumors, respectively. Significant associations were found between high expression of PI3K-p85α, EGFR and p53 and poor prognosis (P = 0.00111; 0.00001; 0.00426). Applying these three proteins as an IHC panel could divide patients into different subgroups (P<0.000001). Multivariate cox regression analysis indicated that the three-protein panel was an independent prognostic factor with very high statistical significance (HR = 2.090, 95% CI: 1.621–2.696, P = 0.00000001). The data suggest that the three-protein panel of PI3K-p85α, EGFR and p53 is an important candidate biomarker for the prognosis of patients with ESCC.     

A Molecular Prognostic Model Predicts Esophageal Squamous Cell Carcinoma Prognosis

Background

Esophageal squamous cell carcinoma (ESCC) has the highest mortality rates in China. The 5-year survival rate of ESCC remains dismal despite improvements in treatments such as surgical resection and adjuvant chemoradiation, and current clinical staging approaches are limited in their ability to effectively stratify patients for treatment options. The aim of the present study, therefore, was to develop an immunohistochemistry-based prognostic model to improve clinical risk assessment for patients with ESCC.

Methods

We developed a molecular prognostic model based on the combined expression of axis of epidermal growth factor receptor (EGFR), phosphorylated Specificity protein 1 (p-Sp1), and Fascin proteins. The presence of this prognostic model and associated clinical outcomes were analyzed for 130 formalin-fixed, paraffin-embedded esophageal curative resection specimens (generation dataset) and validated using an independent cohort of 185 specimens (validation dataset).

Results

The expression of these three genes at the protein level was used to build a molecular prognostic model that was highly predictive of ESCC survival in both generation and validation datasets (P = 0.001). Regression analysis showed that this molecular prognostic model was strongly and independently predictive of overall survival (hazard ratio = 2.358 [95% CI, 1.391–3.996], P = 0.001 in generation dataset; hazard ratio = 1.990 [95% CI, 1.256–3.154], P = 0.003 in validation dataset). Furthermore, the predictive ability of these 3 biomarkers in combination was more robust than that of each individual biomarker.

Conclusions

This technically simple immunohistochemistry-based molecular model accurately predicts ESCC patient survival and thus could serve as a complement to current clinical risk stratification approaches.     

VEGF-A在食管鳞状细胞癌中的表达及临床意义

目的:探讨血管内皮生长因子A(VEGF-A)在食管鳞状细胞癌中的表达及临床意义。方法:收集2009年1月-2010年12月收治的45例食管鳞状细胞癌患者临床资料及病理标本,应用免疫组织化学法检测肿瘤组织VEGF-A表达及微淋巴管密度(MLVD),分析VEGF-A表达与食管鳞癌患者临床病理资料、MLVD及与患者生存期限的关系。结果:1有淋巴结转移的患者VEGF-A表达阳性率为66.67%,明显高于无淋巴结转移患者的38.10%(P0.05);2 VEGF-A阳性食管鳞状细胞癌患者MLVD为(8.35±2.45)明显高于阴性患者的(5.32±1.44),(P0.05);3VEGF-A阳性食管鳞状细胞癌患者3年存活率为41.67%明显低于阴性患者的61.90%(P0.05)。结论:VEGF-A表达在确定早期食管鳞状细胞癌淋巴结转移方面具有一定的应用价值,可以作为评价预后的有效指标。     

ICAM1 Is a Potential Cancer Stem Cell Marker of Esophageal Squamous Cell Carcinoma

Esophageal squamous cell carcinoma (ESCC) accounts for about 90% of esophageal cancer diagnosed in Asian countries, with its incidence on the rise. Cancer stem cell (CSC; also known as tumor-initiating cells, TIC) is inherently resistant to cytotoxic chemotherapy and radiation and associates with poor prognosis and therapy failure. Targeting therapy against cancer stem cell has emerged as a potential therapeutic approach to develop effective regimens. However, the suitable CSC marker of ESCC for identification and targeting is still limited. In this study, we screened the novel CSC membrane protein markers using two distinct stemness characteristics of cancer cell lines by a comparative approach. After the validation of RT-PCR, qPCR and western blot analyses, intercellular adhesion molecule 1 (ICAM1) was identified as a potential CSC marker of ESCC. ICAM1 promotes cancer cell migration, invasion as well as increasing mesenchymal marker expression and attenuating epithelial marker expression. In addition, ICAM1 contributes to CSC properties, including sphere formation, drug resistance, and tumorigenesis in mouse xenotransplantation model. Based on the analysis of ICAM1-regulated proteins, we speculated that ICAM1 regulates CSC properties partly through an ICAM1-PTTG1IP-p53-DNMT1 pathway. Moreover, we observed that ICAM1 and CD44 could have a compensation effect on maintaining the stemness characteristics of ESCC, suggesting that the combination of multi-targeting therapies should be under serious consideration to acquire a more potent therapeutic effect on CSC of ESCC.     

Characterization of Tumor Suppressive Function of cornulin in Esophageal Squamous Cell Carcinoma

By using cDNA microarray analysis, we identified cornulin (CRNN) gene was frequently downregulated in esophageal squamous cell carcinoma (ESCC). In the present study, we investigated the role of CRNN in ESCC development. The results showed that CRNN was frequently downregulated in primary ESCCs in both mRNA level (26/56, 46.4%) and protein level (137/249, 55%), which was significantly associated with lymph node metastases (P=0.027), advanced clinical stage (P=0.039), and overall survival rate (P<0.001). Multivariate analysis indicated that the CRNN downregulation was an independent prognostic factor for ESCC. Functional studies with both in vitro and in vivo assays demonstrated that CRNN had strong tumor suppressive ability in ESCC cells. The tumor-suppressive mechanism of CRNN was associated with its role in cell cycle arrest at G1/S checkpoint by upregulating expressions of P21^WAF1/CIP1 and Rb. Silencing CRNN expression by RNA interference could effectively inhibit its tumor suppressive effect. In conclusion, our findings demonstrate that CRNN is a tumor suppressor gene that plays a critical tumor suppressive role in ESCC.     

高良姜素对人食管鳞癌KYSE-510细胞的抑制作用

探讨在体外高良姜素对人食管鳞癌KYSE-510细胞的抑制作用以及可能的作用机制.MTT结果表明,高良姜素对人食管鳞癌KYSE-510细胞具有很强的生长抑制作用.光学显微镜、激光共聚焦显微镜以及流式细胞仪的分析结果表明,高良姜素可诱导KYSE-510细胞分化.荧光定量RT-PCR和Western印迹分析结果表明,高良姜素诱导P21^waf1、抑制细胞周期蛋白B1和细胞周期蛋白D1的表达,推测上述基因可能是高良姜素实现细胞分化诱导作用的靶基因.     

Oncogenic Features of PHF8 Histone Demethylase in Esophageal Squamous Cell Carcinoma

Esophageal cancer is the sixth leading cause of cancer-related deaths worldwide. It has been reported that histone demethylases are involved in the carcinogenesis of certain types of tumors. Here, we studied the role of one of the histone lysine demethylases, plant homeodomain finger protein 8 (PHF8), in the carcinogenesis of esophageal squamous cell carcinoma (ESCC). Using short hairpin RNA via lentiviral infection, we established stable ESCC cell lines with constitutive downregulation of PHF8 expression. Knockdown of PHF8 in ESCC cells resulted in inhibition of cell proliferation and an increase of apoptosis. Moreover, there were reductions of both anchorage-dependent and -independent colony formation. In vitro migration and invasion assays showed that knockdown of PHF8 led to a reduction in the number of migratory and invasive cells. Furthermore, downregulation of PHF8 attenuated the tumorigenicity of ESCC cells in vivo. Taken together, our study revealed the oncogenic features of PHF8 in ESCC, suggesting that PHF8 may be a potential diagnostic marker and therapeutic target for ESCC.     

GRP78 在食管鳞状细胞癌、不典型增生及正常食管鳞状上皮 组织中的表达

目的:检测葡萄糖调节蛋白78(glucose-regulated proteins 78,GRP78)在同一食管癌患者食管的鳞状细胞癌、不典型增生及正常鳞状上皮组织中的表达,研究GRP78与食管鳞状细胞癌发生发展的关系。方法:取90例食管鳞状细胞癌患者的病变组织,其病理切片中正常鳞状上皮组织、不典型增生组织和鳞状细胞癌组织均存在,用免疫组化的方法 ,检测GRP78在3种不同组织中表达的情况,分析GRP78与性别、年龄、浸润深度、分化程度、分期及淋巴结转移等参数之间的关系。结果:GRP78在食管正常鳞状上皮组织、不典型增生组织、鳞状细胞癌组织中的阳性表达率分别为7.8%、85.6%、47.8%,GRP78在正常食管组织、不典型增生组织、食管鳞癌组织3组中表达的差异有显著性意义(P<0.01)。鳞状细胞癌组织中GRP78表达阴性者,年龄较阳性者大;GRP78表达与肿瘤分化程度、分期、淋巴结转移等有明显相关性。结论:GRP78在食管正常细胞向恶性细胞转化的过程中可能扮演了重要角色,检测GRP78的表达可能有助于对食管鳞状细胞癌的预防及早期诊断。     

GRP78在食管鳞状细胞癌、不典型增生及正常食管鳞状上皮组织中的表达

目的：检测葡萄糖调节蛋白78（glucose—regulated proteins78,GRP78）在同一食管癌患者食管的鳞状细胞癌、不典型增生及正常鳞状上皮组织中的表达,研究GRP78与食管鳞状细胞癌发生发展的关系。方法：取90例食管鳞状细胞癌患者的病变组织,其病理切片中正常鳞状上皮组织、不典型增生组织和鳞状细胞癌组织均存在,用免疫组化的方法,检测GRP78在3种不同组织中表达的情况,分析G史P78与性别、年龄、浸润深度、分化程度、分期及淋巴结转移等参数之间的关系。结果：GRP78在食管正常鳞状上皮组织、不典型增生组织、鳞状细胞癌组织中的阳性表达率分别为7．8％、85．6％、47．8％,GRP78在正常食管组织、不典型增生组织、食管鳞癌组织3组中表达的差异有显著性意义（P〈0．01）。鳞状细胞癌组织中GRP78表达阴性者,年龄较阳性者大;GRP78表达与肿瘤分化程度、分期、淋巴结转移等有明显相关性。结论：GRP78在食管正常细胞向恶性细胞转化的过程中可能扮演了重要角色,检测GRP78的表达可能有助于对食管鳞状细胞癌的预防及早期诊断。