涎腺黏液表皮样癌中p16、p15的表达及相关性研究

[目的]研究p16、p15在正常涎腺组织和黏液表皮样癌中的表达,探讨其在黏液表皮样癌发生、发展中的作用及临床意义.[方法]应用免疫组化S-P法检测10例正常涎腺组织、45例黏液表皮样癌中p16、p15蛋白的表达.[结果]黏液表皮样癌中p16、p15的阳性表达率分别为62.2%、73.3%,显著低于其在正常涎腺组织中的表达(100%)(P＜0.05),且随肿瘤分化程度的降低而递减并具有显著性差异,其中p16的阳性率与淋巴结转移有关.p16、p15两基因在黏液表皮样癌的表达中呈显著相关性(P＜0.05).[结论]p16、p15的失表达与黏液表皮样癌的发生和发展关系密切,p16基因的异常可能是更为直接的因素.     

TGFβ1及其Ⅱ型受体在涎腺黏液表皮样癌中的表达

[目的]研究TGFβ1，TGFβRⅡ在涎腺黏液表皮样癌中的表达和意义。[方法]应用免疫组织化学SP法检测10例正常涎腺组织，45例黏液表皮样癌中TGFβ1和TGFβRⅡ的表达。[结果]正常涎腺组织，黏液表皮样癌中均表达TGFβ1阳性，但正常组的染色强度明显低于肿瘤组（P〈0.05）；TGFβRⅡ在正常涎腺组的阳性表达率为100%，在肿瘤组的阳性表达率为42.22%，两者具有显著性差异（P〈0.05），且高分化组TGFβRⅡ的阳性表达显著高于中，低分化组。[结论]TGFβ1的高表达和TGFβRⅡ的缺失在黏液表皮样癌的发生发展过程中可能起重要作用。     

多药耐药基因在涎腺粘液表皮样癌中的表达及意义

目的探讨涎腺粘液表皮样癌多药耐药基因MDR1/P-gp的表达情况及其临床意义。方法应用免疫组织化学技术(EnVision法)检测36例涎腺粘液表皮样癌(其中高分化组21例、低分化组各15例)以及15例正常涎腺组织中MDR1/P-gp的表达。结果MDR1/P-gp在粘液表皮样癌及正常涎腺组织中均有不同程度表达,阳性部位主要见于细胞膜。高分化粘液表皮样癌、低分化粘液表皮样癌和正常涎腺组织MDR1/P-gp表达阳性率分别为81.0%(17/21)、73.3%(11/15)和26.7%(4/15),组间两两比较均具有显著性差异(P<0.01)。结论MDR1/P-gp是涎腺粘液表皮样癌多药耐药产生的重要细胞机制,检测MDR1/P-gp可为临床拟定化疗方案提供依据,也可作为判断涎腺粘液表皮样癌组织分化程度及预后的参考指标。     

P-gp、GST-π和TopoⅡ在涎腺黏液表皮样癌中的表达及意义

目的：探讨涎腺黏液表皮样癌多药耐药基因P-gp、GST-π和TopoⅡ的表达情况及意义。方法：应用免疫组织化学技术（SP法）检测37例涎腺黏液表皮样癌及12例正常涎腺组织中P-gp、GST-π和TopoⅡ的表达。结果：P-gp、GST-π和TopoⅡ在涎腺黏液表皮样癌及正常涎腺组织中均有不同程度表达。涎腺黏液表皮样癌组织中P-gp、GST-π和TopoⅡ表达阳性率分别为91.9%（34/37）、86.5%（32/37）和21.6%（8/37）。TopoⅡ的表达与肿瘤的分化程度呈负相关。结论：P-gp、GST-π和TopoII在涎腺黏液表皮样癌的多药耐药机制具有重要作用。检测P-gp、GST-π和TopoⅡ可为临床拟定化疗方案提供依据,也可作为判断涎腺黏液表皮样癌预后的参考。     

微波辐射后大鼠肺组织中VEGF和AQP5的表达变化

目的 探讨微波辐射后大鼠肺组织中VEGF和AQP5的表达及其意义。方法 采用10、30和100mW／cm^2的微波辐射96只二级Wistar雄性大鼠，于辐射后6h、1d、3d、7d和14d取肺组织，采用免疫组织化学和图象分析技术探讨血管内皮生长因子（vascular endothelial growth factor，VEGF）和水通道蛋白5（aquaporin5，AQP5）在辐射后大鼠血-气屏障（blood—air barrier）改变中的作用。结果 （1）大鼠肺组织中VEGF表达变化：假辐射组VEGF见于血管内皮细胞浆，呈弱阳性表达。30、100mW／cm^2组微波辐射后6h～7d，VEGF于血管内皮细胞浆呈阳性表达，6h即见增加，1d达高峰，3、7d也呈阳性表达，14d恢复至正常。图象分析结果显示，6h～7d与假辐射组相比有显著性差异（P〈0.05或P〈0.01），且上述改变与辐射剂量呈正相关；（2）大鼠肺组织中AQP5表达改变：AQP5在假辐射组Ⅰ型肺泡上皮细胞膜呈阳性分布，上述三组在辐射后Ⅰ型肺泡上皮细胞膜AQP5的表达均减弱，其中100mW／cm^2组减弱最明显，1d呈弱阳性，3d见恢复，7d基本恢复至正常。结论 10-100mW／cm^2的微波辐射可使VEGF表达增加，AQP5表达减少，并且有时效性和量效性关系，二者可能参与了微波辐射致血．气屏障损伤的病理生理过程。     

乳腺肿瘤组织AQP1表达及其临床意义

目的:探讨水通道蛋白1(AQP1)在乳腺癌组织及乳腺良性肿瘤中的表达特点,以及其在乳腺癌发生、发展中的意义.方法:收集乳腺癌136例、乳腺良性肿瘤68例,应用免疫组织化学方法测定 AQP1在乳腺癌和乳腺良性肿瘤组织中表达水平及分布特点.结果:AQP1在乳腺良性肿瘤细胞及其周围的血管内皮细胞中呈低表达或不表达,但是在乳腺癌腺上皮细胞及乳腺癌组织中的血管内皮细胞呈高表达.两组AQP1阳性细胞的表达率分别为35.66±7.32和277.39±19.35,两者差异有统计学意义,P<0.001.同时乳腺癌组织中AQP1表达及分布存在异质性.结论:AQP1在乳腺良、恶性肿瘤组织中表达存在显著差异,AQP1在乳腺癌组织中呈过高表达,这可能与乳腺癌的恶性生物学行为有关.     

喉部黏液表皮样癌1例并文献复习

目的:探讨喉部黏液表皮样癌的临床病理特征.方法:对1例喉部黏液表皮样癌进行HE切片、免疫组织化学染色等观察并复习文献.结果:镜检肿瘤由黏液细胞、表皮样细胞及中间型细胞组成,免疫组化染色示:CEA(+),EMA(+),CK5/6(+),CK8(+).结论:喉部黏液表皮样癌较为少见,确诊主要依靠组织病理学并辅以免疫组织化学染色,应与鳞状细胞癌、腺鳞癌及腺样囊性癌相鉴别.     

116例腮腺粘液表皮样癌患者的预后因素分析

背景与目的:粘液表皮样癌是涎腺中最常见的恶性肿瘤,有关腮腺的粘液表皮样癌的大宗病例报道较少.本研究旨在探讨影响腮腺粘液表皮样癌患者预后的临床病理因素.方法:回顾性分析中山大学肿瘤防治中心1980年5月至2000年12月收治的116例腮腺粘液表皮样癌患者的临床资料,对其预后进行单因素和多因素分析.结果:116例腮腺粘液表皮样癌患者的5、10和15年生存率分别为75.64%、64.55%和60.39%.单因素生存分析显示年龄、饮酒及T分期等12项因素是腮腺粘液表皮样癌预后的影响因素.多因素分析表明T分期(P=0.006,OR＞1)、病理分级(P=0.000,OR＞1)、远处转移(P=0.000,OR＞1)是影响腮腺粘液表皮样癌患者预后的独立因素.结论:T分期、病理分级和远处转移是影响腮腺粘液表皮样癌患者预后的独立危险因素.     

ER、PTEN在不同类型子宫内膜组织中的表达及意义

目的 检测ER、PTEN在不同类型子宫内膜腺上皮细胞中的表达.方法采用免疫组化检测ER、PTEN的表达.结果⑴ER的表达:在正常子宫内膜腺上皮细胞增生期显著高于分泌期(P＜0.01).增殖症中显著高于正常宫内膜(P＜0.05),而内膜癌中明显低于增殖症和正常宫内膜(P＜0.05).在增殖症中:单纯＞复杂＞不典(均P＜0.05).内膜样腺癌中:＜50岁和≥50岁年龄组、＞1/2和≤1/2肌层浸润组和不同的手术病理分期间ER的表达均无显著性差异(P＞0.05).而在高、中和低分化程度之间均有显著性差异(均P＜0.05);⑵PTEN的表达:在增生期和分泌期无差异(P＞0.05).不典与单纯、不典与复杂相比均有显著性差异(均P＜0.01).内膜样腺癌中:PTEN表达在＜50岁和≥50岁组,不同分化程度的癌组织,不同肌层浸润深度和不同分期之间均无显著性差异(P＞0.05);⑶ER、PTEN在不典型增生和子宫内膜样癌组织中表达均无相关性(r=0.315,r＝0.422,P＞0.05);结论⑴ER参与了正常宫内膜的变化,ER表达降低与内膜样癌的发生有一定关系,且ER表达减少与癌细胞恶性程度增加有关,故可以作为病理诊断的一项指标;⑵PTEN表达降低促使了癌前病变的发生,可能是引起内膜细胞癌变的机制之一,但是PTEN表达与临床病理学特征关系不大;⑶雌激素对PTEN的表达影响不大.     

AQP5与非小细胞肺癌临床特征及转移的关系

[目的]探讨AQP5在非小细胞肺癌(NSCLC)原发灶和淋巴结转移灶中的表达及其意义.[方法]应用免疫组织化学LSAB法检测94例NSCLC原发灶及51例淋巴结转移灶中AQP5的表达情况.[结果]AQP5在腺癌中的表达明显高于鳞癌(P=0.002);在伴有淋巴结转移的NSCLC原发灶中AQP5的表达明显高于不伴有淋巴结转移的原发灶(P=-0.024),而转移灶的表达率与原发灶相比差异无统计学意义(P=0.337);AQP5表达还与NSCLC的TNM分期有关(P=-0.027),在Ⅲ期和Ⅳ期中的表达明显高于Ⅰ期和Ⅱ期.AQP5表达与NSCLC患者的生存率无关(P=0.051).[结论] AQP5表达与NSCLC的发生发展有关,有望成为研究肺癌发生发展和转移的新靶点.     

Inhibition of the aquaporin 3 water channel increases the sensitivity of prostate cancer cells to cryotherapy

Aquaporins (AQPs) are intrinsic membrane proteins that facilitate selective water and small solute movement across the plasma membrane. In this study, we investigate the role of inhibiting AQPs in sensitising prostate cancer cells to cryotherapy. PC-3 and DU145 prostate cancer cells were cooled to 0, −5 and −10°C. The expression of AQP3 in response to freezing was determined using real-time quantitative polymerase chain reaction (RT–qPCR) and western blot analysis. Aquaporins were inhibited using mercuric chloride (HgCl₂) and small interfering RNA (siRNA) duplex, and cell survival was assessed using a colorimetric assay. There was a significant increase in AQP3 expression in response to freezing. Cells treated with AQP3 siRNA were more sensitive to cryoinjury compared with control cells (P<0.001). Inhibition of the AQPs by HgCl₂ also increased the sensitivity of both cell lines to cryoinjury and there was a complete loss of cell viability at −10°C (P<0.01). In conclusion, we have shown that AQP3 is involved directly in cryoinjury. Inhibition of AQP3 increases the sensitivity of prostate cancer cells to freezing. This strategy may be exploited in the clinic to improve the efficacy of prostate cryotherapy.     

AQP1在骨肉瘤中的表达和意义

目的探讨骨肉瘤中AQP1的表达和临床意义。方法对39例骨肉瘤和22例骨良性疾病对照组标本采用免疫组化法观察AQP1的表达并用人工计数和自动图像分析两种方法检测其表达差别,同时行CD34抗体染色标记肿瘤微血管,计算所有标本MVD。结果①AQP1表达于肿瘤微血管和小血管内皮细胞及绝大多数骨肉瘤的肿瘤细胞,尤其见于骨肉瘤的肿瘤巨细胞上;②人工计数AQP1阳性细胞百分比（YP）、AQP1人工计数分级、自动图像分析所得平均光密度（MD）,均表明AQP1在骨肉瘤中高表达（P均〈0.01）;③骨肉瘤中AQP1的YP与MVD呈明显正相关（r=0.341,P〈0.05）。结论大多数骨肉瘤肿瘤细胞高表达AQP1的原因不明;AQP1在骨肉瘤生物学行为及肿瘤微血管生成中占有重要作用;提示AQP1有可能作为骨肉瘤治疗的新靶点。     

Differential expression of aquaporin 5 and aquaporin 3 in squamous cell carcinoma and adenoid cystic carcinoma

Aquaporins (AQPs) are a membrane protein family involved in the selective transport of water across cell membranes. Recent studies have reported the expression of AQP5 in several tumor types such as gastric, pulmonary, ovarian, pancreatic and colorectal cancer. We have previously reported the expression on tumor cells and the important role of AQP3 on cell growth in tongue cancer. However, little is known about the expression and precise role of AQP5 on squamous cell carcinoma (SCC) of the tongue. We investigated the expression of AQP5 and AQP3 in human oral SCC and adenoid cystic carcinoma (ACC). Overexpression of both AQP5 and AQP3 were immunohistochemically observed on tumor cells in SCC, whereas ACC cells were faintly stained with those antibodies against AQPs. Treatment with pan-AQP inhibitor or specific AQP5-siRNA showed inhibition of cell growth in SCC cell lines via the inhibition of integrins and the mitogen-activated protein kinase pathway. AQPs play important roles in cell growth in SCC rather than ACC.     

Overexpression of AQP5, a putative oncogene, promotes cell growth and transformation

Overexpression of several aquaporins has been reported in different types of human cancer but the role of AQPs in human carcinogenesis has not yet been clearly defined. Here, we demonstrate that ectopic expression of human AQP5 (hAQP5), a water channel expressed in lung, salivary glands, and kidney, induces many phenotypic changes characteristic of transformation both in vitro and in vivo. Furthermore, the cell proliferative ability of AQP5 appears to be dependent upon the phosphorylation of a cAMP-protein kinase (PKA) consensus site located in a cytoplasmic loop of AQP5. In addition, phosphorylation of the PKA consensus site was found to be phosphorylated preferentially in tumors. These findings altogether indicate that hAQP5 plays an important role in human carcinogenesis and, furthermore, provide an attractive therapeutic target.     

Aquaporins mediate the chemoresistance of human melanoma cells to arsenite

The integral membrane channel protein aquaporin (AQP) is aberrantly expressed with oncogenic characteristics in various human cancers. In this study, we analyzed the expression pattern of all subtypes of AQPs, and found that 8 out of 13 AQPs expressed in melanoma cells. To understand the role of aberrant expression of AQP in this disease, we over-expressed AQP3 and AQP9 in human melanoma WM266.4 cells and found that both AQPs significantly increased the chemoresistance of WM266.4 cells to arsenite. Functional studies showed that AQP3 and AQP9 can inhibit cell apoptosis induced by arsenite through down-regulating p53 and up-regulating Bcl-2 and XIAP. Our data suggest the implication of APQ in melanoma progression and that the over-expression of AQP3 and AQP9 contributes to the chemoresistance of melanoma to arsenite.     

水通道蛋白3在非小细胞肺癌中的表达及与临床病理相关性的研究

背景与目的肺癌是严重威胁人类生存和发展的恶性疾病之一,本研究旨在探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中水通道蛋白-3(aquaporins3,AQP3)的表达,探讨其与NSCLC临床病理学之间的关系。方法应用免疫组织化学方法检测180例NSCLC组织中AQP3表达及微血管密度(micro vascular density,MVD)。结果 NSCLC组织中AQP3阴性表达率为13.9%(25/180),中等强度阳性表达率为37.2%(67/180),强阳性表达率为48.9%(88/180)。NSCLC组织中AQP3表达高于癌旁组织,有明显统计学差异(P<0.01)。AQP3高表达也同时伴随着MVD计数增高(P<0.01)。男性患者AQP3表达高于女性患者(P=0.003)。腺癌中AQP3的表达较鳞癌明显增强(P<0.001);有淋巴结转移的病例存在AQP3高表达(P=0.026)。NSCLC中AQP3的阳性表达率与肿瘤分化程度呈正相关,表现为AQP3的阳性表达率在高分化癌中明显高于低分化癌(P<0.001)。结论 AQP3在NSCLC的肿瘤血管生成和进展中起重要促进作用,AQP3可能为NSCLC治疗的新靶点。     

水通道蛋白1与恶性肿瘤的相关性

水通道蛋白1(AQP1)是细胞膜上特异性通道蛋白家族中的一类,主要介导水等小分子溶质的跨生物膜转运.现有研究表明,与正常组织对比,AQP1在恶性肿瘤中显著高表达,对肿瘤细胞的增殖、迁移以及血管生成等病理过程产生影响,AQP1有望成为疾病早期筛选、诊断、治疗、判断预后的分子标志.     

Potential role of aquaporin 3 in gastric intestinal metaplasia

Gastric intestinal metaplasia (GIM) is a pre-cancerous condition and a pivotal step in the formation of gastric cancer (GC). Aquaporin 3 (AQP3) has been found to be expressed in goblet cells rather than mucus-secreting glands. To investigate the characteristics of GIM in non-cancerous tissues adjacent to GC, as well as the expression and role of AQP3 in GIM tissues, 16 patients diagnosed with gastric adenocarcinoma of intestinal type located in the lesser curve of the antrum were consecutively enrolled in this study. A new pathological technology called “gastric mucosal sausage roll” was introduced. GIM was determined according to the updated Sydney system, and AQP3 expression in goblet cells was determined by immunohistochemistry. GIM was found in all stomach specimens, and its incidence increased with progression to GC (P < 0.001). GIM prevalence displayed remarkable association with the distance to GC in the anterior gastric wall tissues (P = 0.016) and tissues toward the cardia (P = 0.014), such that GIM was more common in the areas closer to GC (P < 0.001). AQP3 was found to be expressed in 67.71% of parts with GIM, and AQP3 immunoreactivity was identified more frequently in severe GIM areas (P < 0.001). In short, the incidence and severity of GIM correlated with the distance from GC, and AQP3 was differentially expressed in goblet cells, with most AQP3-positive goblet cells presenting in severe GIM. Together, this study suggests that AQP3 may play an important role in gastric carcinogenesis from GIM.     

Development and characterisation of a monoclonal antibody family against aquaporin 1 (AQP1) and aquaporin 4 (AQP4)

Recent studies have discovered the existence of water-channel molecules, the so called aquaporins (AQP) presumably involved in active, ATP dependent water transport between the intracellular and extracellular compartments. Both genetic and protein sequences and structures of the AQPs are known and crystallographic analyses of some members of the AQP family have been performed. Specific antibodies are required to examine their histological locations and analyse their roles in physiological and pathological pathways of water transportation and osmotic regulation. Until recently some polyclonal antibodies have been developed against certain members of the AQP family. However, to date highly specific monoclonal antibodies against aquaporins do not exist. We have developed a monoclonal antibody family against the aquaporin 1 (AQP1) and aquaporin 4 (AQP4) molecules. Well-conserved epitop sequences of AQP1 and AQP4 proteins were selected by computer analysis and their synthetic peptide fragments were used as the antigens of immunisation and the following screening. Antibodies were characterised by immunoserological methods (ELISA, dot-blot and immunoblot), flow cytometry and immunohistochemistry of formaldehyde-fixed and paraffin-embedded tissue samples. RT-PCR tests controlled the specificity of the immune reactions. Our monoclonal antibodies recognised AQP1 and AQP4 in all the techniques mentioned above and apparently they are useful both in various quantitative and qualitative measurements of the expressions of AQP1 and AQP4 in several species (human, rat, mouse, invertebrates, even plants). According to preliminary immunohistochemical studies our monoclonal anti-AQP1 and anti-AQP4 antibodies are appropriate tools of patho-morphological examinations on routine formol-paraffin tissue samples.