膀胱移行细胞癌表皮生长因子及其受体表达和意义

目的 探讨表皮生长因子(EGF)、EGF受体(EGFR)在膀胱移行细胞癌(BTCC)中的表达及意义。方法 用免疫组化ABC法对56例BTCC标本和20例癌旁组织EGF、EGFR进行研究。结果 20例癌旁正常膀胱组织中EGF，EGFR几乎不表达；56例ETCC标本中EGF，EGFR表达明显，与正常组比较有显著性差异(P＜0．01)，不同分期和分级BTCC中的阳性表达率均存在显著性差异(P＜0．05)。结论 EGF，EGFR的检测可以作为判断BTCC预后的一个指标。     

Hedgehog信号通路在胰腺癌中的表达与表皮生长因子受体的关系

目的 探讨胚胎发育信号通路Sonic hedgehog(SHH)在胰腺癌组织中的表达及其与表皮生长因子受体(EGFR)的关系.方法 逆转录-聚合酶链反应(RT-PCR)和Western blot法分别检测胰腺癌组织及癌旁组织中SHH和EGFR的mRNA和蛋白表达.结果 SHH mRNA和蛋白在胰腺癌组织中的阳性率分别为81.6%和79.6%,与癌旁组织比较,差异有统计学意义(P＜0.05).EGFR mRNA和蛋白在胰腺癌组织中的阳性率均为73.5%,与癌旁组织比较,差异有统计学意义(P＜0.05).SHH和EGFR蛋白表达与年龄、肿瘤大小、组织学类型和肿瘤部位等病理因素均无明显相关(P＞0.05),而在不同淋巴结转移状况和TNM分期的病例组中,两者表达差异有统计学意义(P＜0.05).配对资料的Spearman相关分析显示,SHH表达与EGFR表达呈正相关(r=0.232,P＜0.05).结论 SHH和EGFR信号通路在胰腺癌组织中呈活化状态,两者之间的相互作用对胰腺癌的发生发展可能有重要影响.     

结缔组织生长因子在结直肠癌中的表达及临床意义

目的 探讨结缔组织生长因子(connective tissue growth factor,CTGF)在结直肠癌组织中的表达及临床意义.方法 采用免疫组化法检测62例结直肠癌及其对应的癌旁组织中CTGF的表达,并对结果进行统计学分析.结果 CTGF在结直肠癌组织和癌旁组织中表达阳性率分别为61.3%(38/62)和19.4%(12/62),其在结直肠癌组织中的表达明显高于其癌旁组织(P＜0.05).CTGF在结直肠癌组织中的表达与其分化程度、浸润深度及淋巴结转移有关,即癌组织分化程度越低,CTGF表达阳性率越低(P=0.030),随癌组织浸润深度的加深而CTGF表达阳性率明显降低(P=0.032),有淋巴结转移者CTGF的表达阳性率明显低于无淋巴结转移者(P=0.017),而与性别无关(P>0.05).结论 CTGF在结直肠癌的发生、发展过程中可能起重要作用,对指导临床治疗和判断预后有重要意义.     

结直肠癌组织中EGFR、BRAF和K-RasK-Ras基因的表达及其临床意义

目的检测血管内皮生长因子受体(EGFR)、BRAF及K-Ras在结直肠癌组织中的表达情况并探讨其在结直肠癌发病机制中的作用。方法采用免疫组织化学SP法检测136例结直肠癌及其相应癌旁正常结直肠组织中EGFR、BRAF和K-Ras基因的蛋白表达情况,并分析其与结直肠癌患者临床病理特征之间的关系。结果 136例结直肠癌组织中EGFR、BRAF及K-Ras蛋白均有表达,阳性率分别为70.59%(96/136)、50.74%(69/136)及47.48%(66/136),均高于其相应在癌旁正常结直肠组织中的表达阳性率〔分别为35.29%(48/136)、16.18%(22/136)及19.52%(27/136)〕,虽然K-Ras的蛋白表达阳性率在二者之间的差异无统计学意义(P0.05),但EGFR和BRAF在结直肠癌组织中的蛋白表达阳性率均明显高于其在癌旁正常结直肠组织中的表达,差异有统计学意义(P0.05)。EGFR表达与肿瘤的TNM分期、分化程度及淋巴结转移有关(P0.05),BRAF表达与肿瘤的TNM分期有关(P0.05),而K-Ras在结直肠癌组织中的蛋白表达与肿瘤的TNM分期、分化程度及淋巴结转移均无关(P0.05)。相关性分析结果显示EGFR、BRAF及K-Ras基因的蛋白表达之间均无相关性(P0.05)。结论 EGFR、BRAF过表达可能与结直肠癌的发生、发展有关,但K-Ras作用尚不明确,抗EGFR、BRAF靶向治疗对结直肠癌患者可能有益。     

人肝细胞癌中表皮生长因子受体的免疫组化研究

表皮生长因子(EGF)和其受体(EGFR)系统功能失调和肿瘤发生及转移有关，应用免疫组化ABC法．对30例人肝细胞癌癌组织和癌旁肝组织进行了EGFR的表达和比较研究。结果显示EGFR主要定位于细胞膜；HCC癌组织EGFR表达明显低于癌旁肝组织(P<0.05)。讨论了人肝细胞癌癌组织表达EGFR的意义及其和癌基因间的关系。     

表皮生长因子受体和a-连接蛋白在结肠癌中的表达

目的 比较表皮生长因子受体(EGFR)和α-连接蛋白(α-cat)在正常结肠及结肠癌中的表达及其与结肠癌生物学行为的关系.方法 用SP免疫组织化学方法检测20例正常结肠和68例结肠癌组织中EGFR和α-cat的表达.结果 正常结肠EGFR表达阴性,结肠癌组织较正常结肠组织中表达增强(69.1%)(P<0.01);正常结肠中α-cat阳性表达100%,结肠癌组织表达25%,明显降低(P<0.01).EGFR在癌组织中的表达越强,结肠癌的淋巴转移及远处转移率越高(P<0.05),α-cat在癌组织中的表达降低与结肠癌的淋巴转移及远处转移显著相关(P<0.05).结论 α-cat在结肠癌组织表达降低及EGFR在结肠癌组织中表达增强与结肠癌分化、Dukes分期、淋巴转移及远处转移的关系密切,联合检测二者在结肠癌组织中的含量可以帮助评价结肠癌的恶性程度.     

雌、孕激素受体和Her-2阴性乳腺癌的临床病理特征

目的 评价雌、孕激素受体和Her-2表达均阴性乳腺癌(triple-negative breast cancer,TNBC)的临床病理特征及其对预后的影响.方法 500例乳腺癌采用免疫组化筛选TNBC,观察TNBC的临床病理特征,并对其中243例乳腺癌进行临床随访. 结果 500例乳腺癌中TNBC占17.6%(88/500).组织学类型主要为浸润性导管癌(NOS)、化生性癌和髓样癌.组织分级Ⅲ级占72.7%(64/88),高于激素受体阳性组和Her-2高表达组(P=0.000).TNBC中CK5/6阳性率30.7%(27/88),EGFR阳性率34.1%(30/88).TNBC中ERCC1和KIT阳性率分别为28.4%(25/88)和34.1%(30/88),均分别高于激素受体阳性组和Her-2高表达组(P=0.032和P=0.026).TNBC 3年累积生存率为71.5%,低于激素受体阳性组(P=0.021),与Her-2高表达组差异无统计学意义(P=0.474). 结论 TNBC是一类具有高侵袭性病理特征和预后不良的乳腺癌;部分病例表达EGFR和ERCC1.     

甲状腺癌中表皮生长因子受体的表达

为了解甲状腺中表皮生长因子受体的表达意义,应用免疫组化方法检测了８１例甲状腺癌中ＥＧＦＲ的表达,并与甲状腺腺瘤和癌旁正常甲状腺组织相比较。结果：甲状腺癌中４５例ＥＧＦＲ表达阳性,而甲状腺腺瘤及癌旁正常甲状腺组织中均未见ＥＧＦＲ表达。ＥＧＦＲ表达阳性率在甲状腺癌的病理类型,临床分期,浸润程度及淋巴结转移诸因素间接羞匀无统计学意义。     

非小细胞肺癌患者中B细胞淋巴瘤-2、人体第二表皮生长因子受体蛋白表达的比较研究

目的 研究非小细胞肺癌（NSCLC）患者B细胞淋巴瘤-2（Bcl-2）、人体第二表皮生长因子受体（Her-2）蛋白的表达情况及临床意义.方法 应用组织芯片、免疫组化技术检测208例NSCLC患者的肿瘤组织、150例癌旁组织以及17例良性肺疾病肺组织中的Bcl-2、Her-2蛋白的表达情况.结果 ①NSCLC癌组织中Bcl-2蛋白表达率明显高于癌旁组织及肺良性疾病肺组织（ P〈0.05）;Bcl-2蛋白表达水平与NSCLC临床分期及淋巴结转移有相关性（P〈0．05）.②NSCLC癌组织中Her-2蛋白的表达率明显高于癌旁组织及肺良性疾病肺组织（P〈0.05）;Her-2蛋白表达水平与NSCLC分化程度差异有统计学意义（P〈0．05）.结论 ①Bcl-2、Her-2蛋白在NSCLC组织中均高表达,提示Bcl-2及Her-2基因的表达异常与NSCLC的发生发展密切相关;②Bcl-2蛋白表达程度与NSCLC的临床分期及淋巴结转移有关,Her-2蛋白表达程度与NSCLC的分化程度有关,提示两者均与肿瘤恶性生物学行为密切相关.     

细胞周期素E和表皮生长因子受体在乳腺癌中的表达及与预后的关系

目的探讨细胞周期素E(cyclinE)和表皮生长因子受体 (epidermalgrowthfactorreceptor,EGFR)在乳腺癌组织中的表达及其与转移复发和预后的关系。方法应用免疫组织化学方法检测110例乳腺癌组织中cyclinE蛋白和EGFR蛋白的表达水平。结果 110例乳腺癌中cyclinE和EGFR的阳性率均为 5 4 5 5 % ,这两种蛋白在乳腺癌组织中的表达呈正相关 (rs=0 82 3,P =0 0 0 1) ;乳腺癌组织中cyclinE和EGFR表达与肿瘤临床分期 (χ2 =12 86 ,P =0 0 0 5 ;χ2 =14 2 1,P =0 0 0 4 )、组织学分级 (χ2 =8 86 ,P =0 0 0 5 ;χ2 =4 90 ,P =0 0 4 )、淋巴结转移 (χ2 =10 2 2 ,P =0 0 0 1;χ2 =9 6 2 ,P =0 0 0 2 )、ER(χ2 =2 9 87,P =0 0 0 1;χ2 =32 0 8,P =0 0 0 1)、PR(χ2 =19 5 6 ,P =0 0 0 1;χ2 =2 6 92 ,P =0 0 0 1)均相关 ;cyclinE和EGFR阳性表达组的病例有局部复发和远处转移的发生率明显高于阴性表达组的病例 (χ2 =7 33,P =0 0 1;χ2 =7 88,P =0 0 0 5 ) ;cyclinE和EGFR阳性表达病例的平均生存时间和 5年生存率均明显低于阴性表达的病例。结论cyclinE和EGFR的表达可预测乳腺癌的转移和复发 ,是指导临床治疗及估计预后的有意义指标。     

SIRT2 Regulates LPS-Induced Renal Tubular CXCL2 and CCL2 Expression

Sirtuin 2 (SIRT2), a NAD⁺-dependent histone deacetylase, is involved in carcinogenesis and genomic instability and modulates proinflammatory immune responses. However, its role in renal inflammatory injury has not been demonstrated. In this study, we explored the expression patterns of CXCL2 and CCL2 in kidney tissue from Sirt2^−/− and Sirt2^+/+ mice and in mouse proximal tubular epithelial (MPT) cells. CXCL2 and CCL2 were significantly downregulated at both the mRNA and the protein levels in kidneys of LPS-treated Sirt2^−/− mice compared with those of LPS-treated Sirt2^+/+ mice. Furthermore, SIRT2 deficiency ameliorated LPS-induced infiltration of neutrophils and macrophages, acute tubular injury, and decrease of renal function. Supporting these observations, CXCL2 and CCL2 expression levels were lower in MPT cells treated with SIRT2-siRNA than in cells treated with control-siRNA, and adenovirus-mediated overexpression of SIRT2 in MPT cells significantly increased the LPS-induced expression of CXCL2 and CCL2 at the mRNA and protein levels. In addition, SIRT2 interacted with mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1), and SIRT2-knockdown increased the acetylation of MKP-1 and suppressed the phosphorylation of p38 MAPK and c-Jun N-terminal kinase in LPS-treated MPT cells. SIRT2 also regulated p65 binding to the promoters of CXCL2 and CCL2. Taken together, these findings indicate that SIRT2 is associated with expression of renal CXCL2 and CCL2 and that regulation of SIRT2 might be an important therapeutic target for renal inflammatory injury.     

Multidomain synthetic peptide B2A2 synergistically enhances BMP-2 in vitro.

A multidomain, synthetic peptide designated B2A2 synergizes the activity of BMP-2. B2A2 interacts with BMP receptor isoforms, potentiating the action of BMP-2 in activating alkaline phosphatase and triggering Smad and MAPK signaling. B2A2's design permits its delivery as a local surface coating as well as a soluble co-factor, thus broadening potential bioengineering applications. INTRODUCTION: BMP-2 induces osteogenic differentiation and accelerates bone repair. Although BMP-2 inhibitors have been discovered, no BMP-2 mimetics or enhancers that function in the physiological range have yet been found. Here we report that a synthetic peptide designated B2A2, consisting of (1) a BMP receptor-targeting sequence, (2) a hydrophobic spacer, and (3) a heparin-binding sequence, is a positive modulator of recombinant BMP-2. MATERIALS AND METHODS: Cultures of mesenchymal cell lines C2C12 and C3H10T1/2 were given B2A2, recombinant BMP-2, or both. Alkaline phosphatase (ALP) activity was assayed by conversion of paranitrophenol phosphate (PNPP). Signaling through Smad and MAP kinase pathways was monitored by Western blot. Receptor binding was assessed by incubating immobilized B2A2 with soluble recombinant receptor-Fc chimeras and detecting bound receptor by anti-Fc antibody ELISA. Surface coating of medical device materials was done by first dip-coating with silyl-heparin, followed by B2A2. RESULTS AND CONCLUSIONS: Treatment of cells with B2A2 alone marginally increased ALP activity. However, B2A2 plus BMP-2 resulted in 5- to 40-fold augmentation of ALP compared with BMP-2 alone in C3H10T1/2 or C2C12 cells, respectively. This synergistic enhancement was observed over a broad concentration range (4-1000 ng/ml BMP-2). B2A2 interacted directly with BMP receptor isoforms (preferentially to BMPR-Ib and ActivinR-II). In cells, B2A2 + BMP-2 led to a repression of MAP kinase and an increase of Smad activation, consistent with known activation pathways of BMP-2. B2A2 was ineffective when paired with other cytokine/growth factors (basic fibroblast growth factor [FGF-2], TGF-beta1, vascular endothelial growth factor [VEGF]). Simultaneous co-administration was not strictly required. Pulse-chase experiments revealed that temporal separations up to 1 h were still effective. B2A2 was also effective when delivered in a polystyrene- or stainless steel-coated surface through a heparin platform (silyl-heparin) while BMP-2 was added exogenously in solution. These results suggest that B2A2 might promote aggregation of receptor subunits, enabling BMP-2 to activate signaling pathways at effectively lower concentrations. Synthetic multidomain constructs like B2A2 may be useful to accelerate bone repair/deposition through augmentation of endogenous levels of BMP-2 or through local BMP-2 contained in artificial or engineered matrices.     

PGD2-CRTH2 Pathway Promotes Tubulointerstitial Fibrosis

Urinary excretion of lipocalin-type PGD₂ synthase (L-PGDS), which converts PG H₂ to PGD₂, increases in early diabetic nephropathy. In addition, L-PGDS expression in the tubular epithelium increases in adriamycin-induced nephropathy, suggesting that locally produced L-PGDS may promote the development of CKD. In this study, we found that L-PGDS–derived PGD₂ contributes to the progression of renal fibrosis via CRTH2-mediated activation of Th2 lymphocytes. In a mouse model, the tubular epithelium synthesized L-PGDS de novo after unilateral ureteral obstruction (UUO). L-PGDS-knockout mice and CRTH2-knockout mice both exhibited less renal fibrosis, reduced infiltration of Th2 lymphocytes into the cortex, and decreased production of the Th2 cytokines IL-4 and IL-13. Furthermore, oral administration of a CRTH2 antagonist, beginning 3 days after UUO, suppressed the progression of renal fibrosis. Ablation of IL-4 and IL-13 also ameliorated renal fibrosis in the UUO kidney. Taken together, these data suggest that blocking the activation of CRTH2 by PGD₂ might be a strategy to slow the progression of renal fibrosis in CKD.Kidney failure is a public health problem worldwide, with increasing incidence and prevalence, high costs, and poor outcomes. CKD is generally progressive, incurable, and ultimately fatal, although some patients resolve with little or no sequelae. Because current treatment is basically limited to slowing the progression to ESRD using angiotensin-converting enzyme inhibitors and angiotensin II type 1 receptor blockers, more efficient therapies with different or additional modes of action are clearly needed.Regardless of disease etiology, tubulointerstitial fibrosis is the common pathway leading to ESRD in many kidney diseases and is regarded as a prognostic factor for renal function.^1–3 It is noteworthy that some clinical trials are proving that antifibrotic therapies, such as pirfenidone against diabetic nephropathy,⁴ are also effective for CKD. Therefore, elucidating the etiological mechanism underlying renal fibrosis and developing novel therapeutic strategies remains a serious, unmet medical need.Lipocalin-type PGD₂ synthase (L-PGDS) is a secretary protein of the lipocalin superfamily that converts PG H₂, a common precursor of prostanoids, to PGD₂. Because the urinary excretion of L-PGDS increases in the early stage of diabetic nephropathy,^5,6 as well as in patients with essential hypertension without any apparent renal injury,⁷ urinary L-PGDS may be an early diagnostic marker of renal injury in these patients. There is evidence indicating that, in the monkey kidney, L-PGDS is synthesized de novo in the loop of Henle, podocytes, and Bowman’s capsule of the glomeruli.⁸ Furthermore, L-PGDS gene expression in the tubular epithelium was increased in adriamycin-induced nephropathy.⁹ These findings suggest that, under conditions of tubulointerstitial stress, locally produced L-PGDS may be involved in the development of CKD. However, the precise pathophysiological significance of L-PGDS in the kidney remains to be determined.PGD₂ interacts with two receptors, the prostanoid DP₁ receptor and the chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2). Activation of the DP₁ receptor by PGD₂ has been shown to produce vasodilation¹⁰ and bronchodilation.¹¹ Furthermore, the DP₁ receptor is expressed by certain leukocyte populations,^12,13 including dendritic cells, where it controls various functions, including cytokine production. CRTH2 was originally identified as an orphan receptor expressed by Th2 lymphocytes. CRTH2 is not structurally related to the DP₁ receptor and belongs to the family of chemokine receptors. Activation of CRTH2 by PGD₂ plays an important role in allergic inflammation via the recruitment of Th2 lymphocytes and other leukocytes¹⁴ and, perhaps more importantly, by driving the production of the Th2 cytokines IL-4, IL-5, and IL-13.¹⁵