乳腺癌中DEC1与claudin-1的表达及其临床意义

目的：分析DEC1（differentiated embryonic chondrocyte expressed gene 1）和claudin-1在乳腺癌组织中的表达和相关性及其与临床病理因素间的关系。方法：采用免疫组化方法检测DEC1和claudin-1在154例乳腺癌中的表达情况。结果：在154例乳腺癌中 DEC1的表达升高与乳腺癌的高分级（P＝0．002）和淋巴结转移（P＝0．048）相关,而与患者年龄（P＝0．769）、肿瘤大小（P＝0．216）、患者的雌激素水平（P＝0．303）及孕激素水平（P＝0．127）无相关性。而claudin-1的表达缺失与患者的雌激素水平（P＝0．006）和淋巴结转移（P＝0．025）相关,而与患者年龄（P＝0．538）、肿瘤大小（P＝0．801）、肿瘤的分级（P＝0．083）及患者的孕激素水平（P＝0．195）无相关性。并且DEC1在乳腺癌中的表达与claudin-1的表达显著负相关（P＜0．001）。结论：乳腺癌中DEC1的表达与claudin-1的表达负相关,并与肿瘤的高分级相关。     

紧密连接蛋白claudin-1 及ZO-1 在胰腺癌中的表达*

目的：探讨紧密连接蛋白claudin- 1 及ZO- 1 在胰腺癌中的表达及在胰腺癌侵袭转移中的意义。方法：应用免疫组织化学方法检测claudin- 1 和ZO- 1 在胰腺癌不同侵袭位点（与非肿瘤胰腺组织相交界的胰腺癌侵袭前沿区、肿瘤中央区、与周围间质相交界的胰腺癌侵袭前沿区）及淋巴结转移灶和正常胰腺组织中的表达。结果：claudin- 1 与ZO- 1 蛋白正常定位于胰腺腺泡细胞和导管上皮细胞胞膜上,而在胰腺癌组织中,claudin- 1 与ZO- 1 蛋白定位从细胞膜异位至细胞浆或表达缺失,重度异位表达率分别为66.7% 和69.2% 。在与非肿瘤胰腺组织相交界处的胰腺癌组织中,低- 未分化胰腺癌的claudin- 1 重度异位表达率（91.7%）明显高于高- 中分化胰腺癌（55.6% ,P<0.05）;与周围间质相交界处的胰腺癌侵袭前沿区claudin- 1 重度异位表达率（89.7%）高于与非肿瘤胰腺组织相交界处的胰腺癌侵袭前沿区（66.7% ,P<0.05）;淋巴结转移灶重度异位表达率最高（92.3% ,P<0.05）。 同样与周围间质相交界处的胰腺癌侵袭前沿区ZO- 1 重度异位表达率（94.9%）也高于与非肿瘤胰腺组织相交界处的胰腺癌侵袭前沿区（64.2% ,P<0.05）,淋巴结转移灶最高（100% ,P<0.05）;原发胰腺癌组织各个位点中claudin- 1 与ZO- 1 表达呈正相关关系。结论：claudin- 1 和ZO- 1 异位表达对胰腺癌的发生起促进作用;claudin- 1 异位表达与胰腺癌的分化有关;claudin- 1 和ZO- 1 异位表达率增加促进胰腺癌的侵袭与转移。     

E-cadherin、claudin-1在乳腺癌中的表达及临床意义

上皮钙黏蛋白（E—cadherin，E—CD）、紧密连接蛋白claudins（CLDNS）的异常与肿瘤的侵袭转移有着紧密联系，我们对60例原发乳腺癌癌组织、癌旁正常组织中E—cadherin、claudin-1的表达进行检测，以探讨其与乳腺癌临床病理特征的相关性。     

乳腺癌中DECl与claudin-1的表达及其临床意义

目的：分析DECl（differentiat edembryonic chondrocyte expressed gene1）和claudin-1在乳腺癌组织中的表达和相关性及其与临床病理因素问的关系。方法：采用免疫组化方法检测DECl和claudin-1在154例乳腺癌中的表达情况。结果：在154例乳腺癌中DECl的表达升高与乳腺癌的高分级（P=0．002）和淋巴结转移（P=0．048）相关，而与患者年龄（P=0．769）、肿瘤大小（P=0．216）、患者的雌激素水平（P=0．303）及孕激素水平（P=0．127）无相关性。而claudin-1的表达缺失与患者的雌激素水平（P=0．006）和淋巴结转移（P=0．025）相关，而与患者年龄（P=0．538）、肿瘤大小（P=0．801）、肿瘤的分级（P=0．083）及患者的孕激素水平（P=0．195）无相关性。并且DECl在乳腺癌中的表达与claudin-1的表达显著负相关（P〈0．001）。结论：乳腺癌中DECl的表达与claudin-1的表达负相关，并与肿瘤的高分级相关。     

卵巢黏液性肿瘤中claudin-3、claudin-4的表达及其意义

目的 探讨claudin-3、-4在卵巢黏液性囊腺癌的发生、发展过程中的作用.方法 应用免疫组织化学、原位杂交方法检测卵巢上皮、黏液性囊腺瘤、交界性黏液性囊腺瘤、黏液性囊腺癌的石蜡包埋组织中claudin-3、-4和claudin-4 mRNA的表达情况.结果 claudin-3、-4、claudin-4 mRNA在卵巢上皮及卵巢黏液性囊腺瘤中表达最弱;交界性黏液囊腺瘤中表达明显增强,却明显弱于黏液性囊腺癌(P<0.05);在低分化卵巢癌中的表达强于高分化癌及中分化癌(P<0.05),中分化癌与高分化癌中的表达无统计学意义(P>0.05).结论 claudin-3、-4表达增强,与黏液性囊腺癌的发生、发展密切相关,与其分化程度有关,可能会成为诊断卵巢黏液性囊腺癌的特异性标记物.     

E-cadherin、claudin-1联合检测与乳腺癌侵袭转移关系的研究

目的:研究上皮钙黏蛋白(E-cadherin,E-cd)、紧密连接蛋白(claudin-1,CLND)在乳腺癌组织中的表达,探讨E-cd、claudin-1与乳腺癌侵袭转移的关系及临床应用中的意义。方法:应用免疫组织化学染色法,检测60例乳腺癌术后标本的癌组织中E-cd、claudin-1的表达,分析两者联合检测与淋巴结转移及癌基因(CerbB2)的关系。结果:E-cd与claudin-1在乳腺癌中的表达无相关性,但在联合检测时,E-cd(-)/claudin-1(-)组淋巴结受累率80%(3/12例)、CerbB2阳性表达率73.3%(11/15例),明显高于E-cd( )/claudin-1( )组25.5%(4/13例)和31.2%(5/11例),有显著性差异(P<0.05)。结论:E-cd、claudin-1在乳腺癌中的表达无相关性,是相对对立的因素,但两基因联合检测是筛选术后转移高危人群可靠的手段;E-cd、claudin-1在抑制肿瘤侵袭转移中起着重要作用,可作为判断预后的参考指标之一。     

Low expression of claudin-4: an indicator of recurrence in esophageal squamous cell carcinoma after Ivor Lewis esophagectomy?

The high recurrence rate after surgery is the main reason for the poor prognosis of esophageal squamous cell carcinoma (ESCC) patients. Finding indicators of recurrence and taking adjuvant therapy may be useful for patients in high risk of recurrence. claudin-4 (CLDN4) is the core protein to form the tight junction, which plays an important role in cell adhesion, and its aberrant expression were detected in various cancers while its expression and functions in ESCC still remained unclear. Here, we detected the expression of CLDN4 in 114 ESCC tissue samples by real-time RT-PCR and immunohistochemistry, and the result showed that the low expression of CLDN4 correlated with a higher T staging (P = 0.010), lymphatic metastasis (P < 0.001) and recurrence status (P = 0.002). And the Cox regression analysis showed that the T classification (P = 0.005), lymph node metastasis (P = 0.003) and low CLDN4 expression (P = 0.029) were independent risk factors of recurrence. Further, we proved the CLDN4 in inhibiting growth, colony formation and invasion in vitro by establishing two stable CLDN4-silencing ESCC cell lines. In conclusion, CLDN4 played an important role in preventing metastasis and could be an effective biomarker to predict the recurrence of ESCC.     

claudin-4和MMP-2在子宫内膜癌中的表达及其意义

目的:探讨claudin-4和MMP-2在子宫内膜癌中的表达与临床病理参数之间关系.方法:采用免疫组化S-P法检测75例子宫内膜癌和28例正常增生期子宫内膜中claudin-4和MMP-2表达并作相应分析.结果:子宫内膜癌中claudin-4 和MMP-2表达均高于正常增生期子宫内膜中的表达(P<0.05);claudin-4和MMP-2高表达均与癌组织肌层浸润深度有关(P<0.05),与组织学类型、组织分级、有无淋巴结转移及手术病理分期无关(P>0.05);claudin-4与MMP-2呈正相关(r= 0.53,P<0.05).结论:claudin-4和 MMP-2高表达可能促进子宫内膜的发生和发展,claudin-4和 MMP-2表达可能是判断子宫内膜癌预后的重要参考指标.     

claudin-4和MMP-2在子宫内膜癌中的表达及其意义

目的：探讨claudin-4和MMP-2在子宫内膜癌中的表达与临床病理参数之间关系。方法：采用免疫组化S—P法检测75例子宫内膜癌和28例正常增生期子宫内膜中claudin-4和MMP-2表达并作相应分析。结果：子宫内膜癌中claudin-4和MMP-2表达均高于正常增生期子宫内膜中的表达（P〈0．05）;claudin-4和MMP-2高表达均与癌组织肌层浸润深度有关（P〈0．05）,与组织学类型、组织分级、有无淋巴结转移及手术病理分期无关（P〉0．05）;claudin-4与MMP-2呈正相关（r=0．53,P〈0．05）。结论：claudin-4和MMP-2高表达可能促进子宫内膜的发生和发展,claudin-4和MMP-2表达可能是判断子宫内膜癌预后的重要参考指标。     

大鼠C6胶质瘤血脑屏障超微结构与紧密连接蛋白claudin-5变化的研究

背景与目的：开放血脑屏障将为胶质瘤的综合治疗提供新的希望,本研究观察C6脑胶质瘤不同区域血-脑屏障（BBB）的通透性的变化、超微结构特征及紧密连接蛋白claudin-5的表达变化。方法：雄性SD大鼠,随机分为对照组和荷瘤组,采用立体定向方法在大鼠右侧尾状核接种C6细胞建立大鼠C6胶质瘤模型,于建模后第20天取肿瘤、肿瘤临近处（brain adjacent to tumor,BAT）即距肿瘤边界2mm以内及肿瘤远隔处（brain dis-tant to tumor,BDT）即2mm以外的大脑半球脑皮质3个部位的脑组织和对照鼠右侧尾状核正常脑组织,采用外源性硝酸镧透射电镜示踪法,观察BBB/BBTB超微结构特征;免疫组化法观察紧密连接蛋白claudin-5在正常鼠和荷瘤鼠各部位的表达变化。结果：（1）肿瘤处可见La^3＋通过血管内皮细胞的紧密连接渗透至血管腔外及脑间质裂隙内,BAT部分见La^3＋渗出至局部基底膜,大部分分布在血管腔内,而BDT及对照组正常脑组织La^3＋分布在血管腔内,血管内皮细胞紧密连接结构完整,基底膜呈线状连续,脑血管内皮细胞浆均未见La^3＋。（2）claudin-5在BDT与对照组脑组织表达丰富、连续;肿瘤处表达减弱,沿血管呈间断表达;BAT较正常脑组织表达下降,但仍呈连续状态。结论：正常脑组织血脑屏障稳定,通透性最低;肿瘤的周边存在不同程度的屏障;C6胶质瘤细胞的直接浸润可以降低内皮细胞紧密连接蛋白claudin-5,可能是诱导紧密连接破坏导致BBTB通透性增高的重要原因。     

Deoxypyrimidine-induced inhibition of the cytokinetic effects of 1-β-d-arabinofuranosyluracil

Summary Ara-U-induced S-phase accumulation and the interaction between high concentrations of ara-U (HiCAU) and ara-C were investigated in L1210 leukemia cells in vitro. Treatment of exponentially growing L1210 murine leukemia cells with ara-U (200–1000 m) for 48 h caused a dose-dependent accumulation of cells in the S-phase. The extent of this ara-U-induced S-phase accumulation correlated with ara-U incorporation into DNA and with increases of up to 172% and 464% in the specific activities of deoxycytidine kinase and thymidine kinase, respectively, over control values. Metabolism of 1 m ara-C following the exposure of cells to ara-U (1mm) resulted in 4.5 pmol ara-C DNA/mg protein vs 2.1 pmol/mg protein in control cells. Although 48-h exposure of cells to 200 and 400 m ara-U is not cytotoxic, it enhances the cytotoxicity of ara-C (10–100 m) 4- to 10-fold. Ara-U-induced S-phase accumulation is inhibited by deoxypyrimidine nucleosides but not by pyrimidine or deoxypurine nucleosides. Some of the ara-U and ara-C concentrations used in this study are achievable in clinical practice, and ara-U/ara-C interactions may explain in part the unique therapeutic utility of high-dose ara-C.Abbreviations ara-C 1--d-arabinofuranosylcytosine - ara-U 1--d-arabinofuranosyluracil - ara-CTP 1--d-arabinofuranosylcytidine triphosphate - HiDAC high-dose ara-C - HiCAU high concentrations of ara-U - dCTP deoxycytidine triphosphate - HiDAU high-dose ara-U - FiTC Fluoroisothiocyanate - dUDP deoxyuridine diphosphate - dUTP deoxyuridine triphosphate - dTTP thymidine triphosphate - BrdUrd bromodeoxyuridine - dCyd kinase deoxycytidine kinase Supported in part by grant CH-35H from the American Cancer Society, by Public Health Service grant CA-12197 from the National Cancer Institute, National Institutes of Health, and by the Gaston Cancer Society     

Expression of TGFβ-1 and EHD1 correlated with survival of non-small cell Lung cancer

Transforming growth factor-β1 (TGFβ-1) signaling is regulated by endocytotic pathway. To clarify the prognostic value of TGFβ-1 and to verify the involvement of endocytosis in drug resistance, we examined the expression of TGFβ-1 and Eps15 homology domain 1 (EHD1) in non-small cell lung cancer (NSCLC) and its association with tumor characteristics and survival of patients with NSCLC. Expression of TGFβ-1 and EHD1 was evaluated by immunohistochemistry in paraffin sections from 105 NSCLC patients. Overall survival (OS) was analyzed by Kaplan–Meier method, log-rank test, and multivariate Cox proportional hazard regression model. Positive immunostaining of TGFβ-1 and EHD1 was detected in 52.38 and 39.05 % of NSCLC samples, respectively. In non-adjuvant chemotherapy-treated group (P?=?0.006) and epidermal growth factor receptor (EGFR) (+) group (P?=?0.038), patients with TGFβ-1 expression had a longer OS. EHD1 negative expression predicted a longer OS (P?=?0.003), especially in EGFR (+) (P?=?0.006) and adjuvant chemotherapy-treated patients (P?=?0.003). NSCLC patients with concurrent positive TGFβ-1 and negative EHD1 (combined markers) were significantly correlated with better OS (P?=?0.001). American Joint Committee on Cancer (AJCC) status and combined markers were independent prognostic indicators for OS (HR (95 % CI) 1.576 (1.112–2.232), P?=?0.011 and HR 0.349 (0.180–0.673), P?=?0.002, respectively). We identified concordant TGFβ-1 positive and EHD1 negative as a strong favorable prognosis factor in NSCLC. Our results may help us to select and optimize strategies for individualized therapy.     

Loss of p53 enhances the function of the endoplasmic reticulum through activation of the IRE1α/XBP1 pathway

Altered regulation of ER stress response has been implicated in a variety of human diseases, such as cancer and metabolic diseases. Excessive ER function contributes to malignant phenotypes, such as chemoresistance and metastasis. Here we report that the tumor suppressor p53 regulates ER function in response to stress. We found that loss of p53 function activates the IRE1α/XBP1 pathway to enhance protein folding and secretion through upregulation of IRE1α and subsequent activation of its target XBP1. We also show that wild-type p53 interacts with synoviolin (SYVN1)/HRD1/DER3, a transmembrane E3 ubiquitin ligase localized to ER during ER stress and removes unfolded proteins by reversing transport to the cytosol from the ER, and its interaction stimulates IRE1α degradation. Moreover, IRE1α inhibitor suppressed protein secretion, induced cell death in p53-deficient cells, and strongly suppressed the formation of tumors by p53-deficient human tumor cells in vivo compared with those that expressed wild-type p53. Therefore, our data imply that the IRE1α/XBP1 pathway serves as a target for therapy of chemoresistant tumors that express mutant p53.     

Deletion of exons 1–3 of the MEN1 gene in a large Italian family causes the loss of menin expression

Multiple endocrine neoplasia type 1 (MEN1) syndrome is an autosomal dominant disease, characterized by parathyroid adenomas, endocrine gastroenteropancreatic tumors and pituitary adenomas, due to inactivating mutations of the MEN1 gene (chromosome 11q13). MEN1 mutations are mainly represented by nonsense, deletions/insertions, splice site or missense mutations that can be detected by direct sequencing of genomic DNA. However, MEN1 patients with large heterozygous deletions may escape classical genetic screening and may be misidentified as phenocopies, thereby hindering proper clinical surveillance. We employed a real-time polymerase chain reaction application, the TaqMan copy number variation assay, to evaluate a family in which we failed to identify an MEN1 mutation by direct sequencing, despite a clear clinical diagnosis of MEN1 syndrome. Using the TaqMan copy number variation assay we identified a large deletion of the MEN1 gene involving exons 1 and 2, in three affected family members, but not in the other nine family members that were to date clinically unaffected. The same genetic alteration was not found in a group of ten unaffected subjects, without family history of endocrine tumors. The MEN1 deletion was further confirmed by multiplex ligation-dependent probe amplification, which showed the deletion extended from exon 1 to exon 3. This new approach allowed us to correctly genetically diagnose three clinical MEN1 patients that were previously considered as MEN1 phenocopies. More importantly, we excluded the presence of genetic alterations in the unaffected family members. These results underline the importance of using a variety of available biotechnology approaches when pursuing a genetic diagnosis in a clinically suggestive setting of inherited endocrine cancer.     

Polymorphic variation of CYP1A1 is associated with the risk of gastric cardia cancer: a prospective case–cohort study of cytochrome P-450 1A1 and GST enzymes

Objective: To determine if genetic polymorphisms of CYP1A1, GSTM1, GSTP1, or GSTT1 are associated with an increased risk of developing esophageal squamous cell carcinoma (ESCC), gastric cardia cancer (GCC), or either in a high-risk Asian population. Methods: We conducted a case-cohort analysis with 5 years of prospective follow-up. The analytical cohort contained 642 individuals who participated in either the Dysplasia Trial (DT) or the General Population (GPT) of the Nutrition Intervention Trials conducted in Linxian, China, and included 131 cases of ESCC and 90 cases of GCC. Genotyping analysis was performed on DNA extracted from red blood cells using a PureGene kit (Gentra Systems, Inc., Minneapolis, MN) and real-time PCR analysis amplification (Taq-Man). Relative risks and 95% confidence intervals were estimated using the case – cohort estimator for the Cox proportional hazards models. p-values from nested models with genotyping variables came from score tests. Results: The relative risks for developing ESCC, GCC, or either cancer were calculated in the entire analytic cohort for GSTM1, P1*B (A313G), and T1 and CYP1A1*2A (T3801C) and *2C (A2455G) genotypes, and no significant associations were identified. However, because of the difference in cancer risks between the DT (9.3 cases per 1000 person years) and the GPT (5.3 cases), the analytical cohort was stratified by trial; the DT participants who were heterozygous or homozygous for the variant-allele at CYP1A1*2A had a reduced risk for developing GCC (adjusted RR (95%CI) 0.47 (0.23–1.00) p = 0.037). Conclusions: This study found an association for the CYP1A1*2A variant allele and a reduced risk of GCC in people at high risk for development of this disease. This finding is consistent with previous studies suggesting that substrates for the cytochrome P-450 1A1 metabolic pathway, such as polycyclic aromatic hydrocarbons, may be etiologically significant in this high-risk region.     

claudin-6及clusterin在胃癌组织及胃癌细胞株中表达的初步研究

目的:探讨claudin-6及clusterin在胃癌组织及胃癌细胞中的表达及其临床意义。方法:应用免疫组织化学及免疫细胞化学法检测claudin-6及clusterin在正常胃黏膜、非典型增生胃黏膜、胃癌组织及胃癌细胞株(MKN28、SGC7901、BGC823)中的表达。结果:胃癌组织、非典型增生胃黏膜及正常胃黏膜中claudin-6及clusterin的阳性表达率分别是53.3%、65%、100%及60%、75%、100%,二者的阳性表达率在三组间有统计学差异(均P<0.05)。在不同浸润深度及有无淋巴结转移组内claudin-6及clusterin阳性表达的差异性有统计学意义(P<0.05),而在不同性别、年龄及脉管侵犯组内无统计学差异(P>0.05),Spearman等级相关性分析显示claudin-6与clusterin表达呈正相关(r=0.899,P<0.05)。在三株胃癌细胞中claudin-6与clusterin均阳性表达在细胞膜和细胞质中。依分化程度clusterin在MKN28、SGC7901、BGC823三种细胞系中免疫染色强度逐步减弱,claudin-6在三种细胞系中免疫染色强度无明显差别。结论:claudin-6与clusterin的缺失与胃癌发生发展及浸润转移有关,有望成为反映胃癌生物学行为有价值的标志物。