Survivin在膀胱移行细胞癌中的表达及其临床意义

目的:检测抗凋亡(IAP)家族中生存素(Survivin)基因在膀胱移形细胞癌(BTCC)组织及癌旁组织的表达,探讨Survivin的表达在膀胱癌发生发展中的意义.方法:采用免疫组织化学和实时荧光定量逆转录聚合酶链反应(RT-QPCR)方法,对30例BTCC患者中Survivin基因在癌组织和癌旁组织中的表达进行检测.结果:免疫染色标本中,在癌旁组织、BTCC和胚胎癌组织中Survivin的阳性表达率分别为0、60%和100%.同一患者,Survivin在BTCC的表达量远大于癌旁组织,Survivin在BTCC组织中的-△△CT值是癌旁组织的10.2829(9.0034～11.5624)倍,同时在病理分级之间和临床分期之间均有统计学意义(P＜0.05).结论:Survivin基因在癌旁组织中有少量表达,而在BTCC组织中的表达量远远高于癌旁组织;其表达量与病理分级和临床分期均有相关性.     

凋亡抑制因子Livin在膀胱移行细胞癌组织中的表达及意义

目的：研究凋亡抑制蛋白家族(IAP家族)新的凋亡抑制因子Livin在膀胱移行细胞癌(BTCC)中的表达及其与肿瘤分级、分期、复发之间的关系。方法：采用免疫组织化学SP法,对54例BTCC和8例正常膀胱组织中Livin的表达情况进行检测,分析其在膀胱癌组织和非膀胱癌组织中的表达及其与肿瘤病理学分级、临床分期和患者复发情况的关系。结果：Livin在8例正常膀胱组织中均不表达,而在54例膀胱移行细胞癌组织中表达率为85．2%(P＜0．01)。Livin的表达和膀胱移行细胞癌的病理分级、临床分期、是否复发无明显相关(P＞0．05)。结论：细胞凋亡抑制因子Livin在BTCC组织中表达上调,提示Livin可能通过抑制细胞凋亡,对BTCC的发生发展起重要作用;Livin在膀胱癌中的高表达有望成为一种有效、敏感的瘤标,并为BTCC的基因治疗提供新的靶点。     

人丝氨酸蛋白酶抑制因子HAI-1在膀胱移行细胞癌中的表达及意义

目的 检测HAI-1基因在膀胱移行细胞癌(BTCC)中的表达,探讨与BTCC临床病理参数间的关系及其临床意义.方法 应用免疫组化SP法检测HAI-1在BTCC及正常膀胱黏膜中的表达,并结合临床病理资料分析.结果 HAl-1在BTCC及正常膀胱组织中均有表达;在BTCC中的表达阳性率明显低于正常膀胱组织,差异有统计学意义(P＜0.001);HAI-1表达随肿瘤病理分级、临床分期的升高、肿瘤转移的发生而显著性降低(P＜0.001).结论 HAI-1异常表达在BTCC的发生发展过程中可能具有重要作用,HAI-1蛋白可作为判断BTCC恶性程度及预后的新型生物学标记.     

HER-2和MGMT在膀胱移行细胞癌中的表达及意义

目的 探讨HER-2和O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methylguanine-DNA-methyltransferase,MGMT)在膀胱移行细胞癌(BTCC)中的表达及意义.方法 采用免疫组织化学法检测80例BTCC组织及30例癌旁组织和20例正常膀胱黏膜组织中HER-2和MGMT的表达,分析其与病理分级、临床分期及其他临床因素的相关性及意义.结果 HER-2在正常膀胱组织和癌旁组织及BTCC中的阳性表达率依次增高.差异有统计学意义(P<0.001),随着BTCC病理分级和临床分期的不同其表达依次增高,差异有统计学意义(P<0.05);MGMT在正常膀胱组织和癌旁组织及BTCC中的阳性表达率依次降低,差异有统计学意义(P<0.001),随着BTCC病理分级和临床分期的不同其表达依次降低,差异有统计学意义(P<0.05).HER-2和MGMT与患者性别、年龄、肿瘤数量及发病次数均无明显相关性(P>0.05).结论 HER-2和MGMT可能成为膀胱癌诊断及判断预后的标志物.     

miR-126在膀胱移行细胞癌中的表达及其与VEGF的相关性

目的研究miR-126在膀胱移行细胞癌（BTCC）中的表达和临床意义,以及与血管内皮生长因子（VEGF）蛋白表达的相关性,探讨miR-126在BTCC发生、发展和转移中的作用及机制。方法采用组织芯片平台,应用原位杂交方法（ISHH）检测56例BTCC组织、41例相应的癌旁组织和15例正常膀胱组织的miR-126表达,免疫蛋白印迹法（Western blotting）检测VEGF蛋白的表达。结果 miR-126在56例BTCC组织中的阳性表达率为23.21%,均显著低于癌旁组织和正常膀胱组织（43.90%和53.33%,P均〈0.05）。VEGF蛋白在BTCC组织中的表达均显著高于癌旁组织和正常膀胱组织。BTCC组织中miR-126的低表达与肿瘤的淋巴转移密切相关,与病理分级有一定关系;而与患者性别、年龄、临床分期、肿瘤直径和肿瘤发生情况等均无相关性。相关性检验表明miR-126与VEGF表达呈显著负相关（P〈0.05）。结论 miR-126在BTCC组织中的表达下调,可促进膀胱癌内微血管形成,提示miR-126可能作为重要的抑癌基因参与BTCC的发生和发展过程,并可能是膀胱癌转移的潜在标志。     

血小板源性生长因子-D在膀胱癌中的表达及其临床意义

目的 观察血小板源性生长因子-D(PDGF-D)在膀胱移行细胞癌(BTCC)中的表达,探讨PDGF-D与膀胱移行细胞癌的发生发展之间的关系及其临床意义.方法 分别采用实时定量逆转录-聚合酶链反应(real-time RT-PCR)法和蛋白免疫印迹(Western blot)法检测62例膀胱移行细胞癌组织、10例癌旁正常膀胱组织中PDGF-D mRNA和蛋白的表达,并分析其与临床病理之间的关系.结果 PDGF-D mRNA在BTCC中的表达量是正常对照组的1.89倍,PDGF-D蛋白在癌组织中的表达明显高于正常膀胱组织(相对吸光度比值:正常组0.231±0.041,癌症组0.689±0.083,P＜0.05).而且随着肿瘤病理分级的升高及淋巴结转移的出现,PDGF-D mRNA和蛋白的表达水平均逐渐升高,各组问差异有统计学意义(P＜0.05).但不同分期的癌组织中PDGF.D mRNA和蛋白的表达水平的差异无统计学意义(P＞0.05).结论 PDGF-D表达增强与膀胱癌的发生及分化、转移密切相关,可能在膀胱癌的发生发展中发挥重要作用.     

膀胱癌中受体酪氨酸激酶EphA2的表达及其意义

目的 探讨EphA2在膀胱移行细胞癌(BTCC)中的表达及其临床意义.方法 采用免疫组织化学SP法检测49例BTCC及10例癌旁正常组织中EphA2的蛋白表达,应用实时定量逆转录-聚合酶链反应(real-time RT-PCR)法检测各临床标本相应的mRNA表达并通过2-△△CT方法分析相对基因表达的差异.结果 相对于癌旁正常组织而言,EphA2在BTCC中的蛋白表达显著增强(积分吸光度IA均值:正常组8364±3915,癌症组19 122±5696,P＜0.01),其mRNA表达平均上调约2.57倍;除中分化(G2)与低分化(G3)组间EphA2 mRNA表达差异无统计学意义(P＞0.05)外,EphA2在其余各分级与分期组间的蛋白及核酸表达,差异均具有统计学意义(P＜0.05);EphA2的表达与膀胱癌的临床病理特征密切相关.结论 EphA2表达增强可能促进了膀胱癌的恶性晋级,EphA2可作为监测膀胱癌恶性发展的指标和治疗的新靶点.     

金属硫蛋白在肝细胞癌中表达的临床意义

目的 探讨金属硫蛋白（metallothionein,MT)在肝细胞癌（hepatocellular carcinoma,HCC)组织中、癌旁组织和正常肝组织中的表达及其临床意义。方法 对30例HCC病人的癌组织、10例癌旁组织和10例正常肝组织标本进行免疫组化染色，观察MT的表达状况，分析MT在HCC中的表达与癌旁组织、正常肝组织、细胞分化和预后等的关系。结果 正常肝组织有很强而稳定的MT染色；癌旁组织（包括肝硬化结节）有较高的MT着色；癌组织中MT染色很弱，甚至无MT染色。相互比较有显著性差异（P＜0．05）。MT在肝细胞癌组织中的表达与生存率之间存在着正相关关系（P＜0．05）。结论 MT在正常肝组织、癌旁组织和肝细胞癌组织中的表达呈递减趋势，肝细胞癌组织中MT表达与癌细胞分化无关，而与瘤体组织坏死及肝切除术后病人生存状况相关。     

透明质酸在膀胱移行细胞癌、乳头状瘤组织中的表达及其意义

目的　探讨人膀胱移行细胞癌 (BTCC)、乳头状瘤组织中透明质酸的表达情况及其意义。方法　应用免疫组织化学染色方法检测 60例BTCC、2 5例膀胱乳头状瘤和 15例正常膀胱黏膜上皮组织标本中透明质酸表达情况并进行统计学分析。结果　正常对照组HA染色 :(-) 11例 ,( ) 4例 ,染色位于上皮细胞间质。膀胱乳头状瘤组染色 :(-) 3例 ,( ) 10例 ,( ) 12例 ;G1级膀胱移行细胞癌组染色 :(-) 2例 ,( ) 7例 ,( ) 11例 ;G2 级 :( ) 2例 ,( ) 13例 ,( ) 5例 ;G3级 :( ) 1例 ,( ) 8例 ,( ) 11例 ;染色均位于癌细胞胞浆和间质中。正常对照组与BTCC、膀胱乳头状瘤组 ,膀胱乳头状瘤组与BTCC组比较 ,差异均有极显著性 (P <0 .0 0 1) ;G1级组与G2 级组、G1级组与G3 级组之间差异有显著性 (P =0 .0 13 2 ,0 .0 0 3 5 ) ,G2 级组与G3 级组比较 ,差异无显著性 (P =0 .0 5 2 8) ;膀胱乳头状瘤组与G1级组之间的差异无显著性 (P =0 .64 0 7)。结论　透明质酸与BTCC的生物学特性有直接的关联 ,膀胱移行细胞癌细胞本身具有合成透明质酸的功能 ;膀胱乳头状瘤的生物学行为有恶性倾向     

凋亡抑制因子Livin在膀胱移形细胞癌中的表达

目的检测抗凋亡（IAP）家族中Livin基因在膀胱移形细胞癌（BTCC）组织及癌旁组织的表达，探讨Livin的表达在膀胱癌发生发展中的意义。方法采用免疫组织化学和实时荧光定量逆转录．荧光定量聚合酶链反应（RT-QPCR）方法对30例膀胱癌患者中Livin基因在癌组织和癌旁组织中的表达进行检测。结果免疫染色标本中，在癌旁组织和膀胱癌组织中Livin的阳性表达率分别为0．60％。Livin在膀胱癌组织中的-△△CT值是癌旁组织的8．0454（7．4264—8．6644）倍，与分级和分期没有相关性。结论Livin基因在癌旁组织中有少量表达，而在BTCC组织中的表达量远远高于癌旁组织。     

Clinical significance of EGF and EGFR expression changes in cryptorchid boys

Aim: To explore the changes of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) expressions in cryptorchid children and their clinical significance. Methods: The level of serum EGF was measured by radioimmunoassay (RIA) and the expression of EGFR by immunohistochemistry. Results: (1) The level of serum EGF was significantly lower in cryptorchid children than in normal subjects at age group of 59 years (P<0.01) and 10-14 years (P<0.01), (2) The level of EGF was significantly lower in boys with impalpable testis than in those with extracanalicular and intracanalicular testes (P<0.01), (3) The serum EGF level increased significantly 6 months after orchiopexy (P<0.05), (4) The EGFR expression in testicular Leydig's cells was lower in 2-4 year-old boys than in those over 5 years (P<0.05) and (5) the EGFR expression was less positive in the impalpable group and the intracanalicular group than that of the extracanalicular group (P<0.01). Conclusion: The EGF and EGFR expr essions may co     

Expression of the epidermal growth factor receptor family in normal and malignant urothelium

OBJECTIVE: To compare the immunohistochemically assessed expression of the epidermal growth factor receptor (EGFR) family in normal and malignant bladder urothelium, and suggest new hypotheses about their function in the development and progression of transitional cell carcinoma (TCC). PATIENTS AND METHODS: EGFR, ERBB2, ERBB3 and ERBB4 were evaluated immunohistochemically in normal urothelium (NU, 15), primary non-metastasized invasive TCC (NMC, 19) and in primary invasive TCCs with corresponding metastases (MC, 51, both specimens). RESULTS: All NU samples expressed ERBB4, none expressed ERBB2 and two expressed EGFR; all staining was uniform throughout all cell layers. ERBB2 expression increased and ERBB4 decreased from normal samples to carcinomas. There was no difference between NMCs and MCs in ERBB2, ERBB3 and ERBB4, but the NMCs expressed more EGFR than both NU and MC samples. There were no associations with T category, grade or survival. All combinations of expression levels for the four receptors were detected, with no dominant profile. CONCLUSION: We hypothesise that: (i) ERBB4 is important for differentiation in NU; (ii) ERBB2 is up-regulated with carcinogenesis in the urinary bladder but does not discriminate between bladder cancer with or without metastases; (iii) EGFR may be a marker of indolent disease. A current hypothesis, that superficial layers of NU do not express EGFR and thus protect the basal cells from the mitogenic effect of urinary EGF, is challenged.     

Changes in epidermal growth factor receptor expression in human bladder cancer cell lines following interferon-alpha treatment

PURPOSE: We examined the regulation of epidermal growth factor (EGF) receptor (EGFR) expression in human bladder cancer cell lines by interferon-alpha (IFN-alpha), the ability of IFN-alpha to inhibit cell proliferation and the sensitivity of IFN-alpha pretreated cells to EGF. MATERIALS AND METHODS: Cell proliferation was determined using crystal violet colorimetric and clonogenic assays. EGFR expression was measured by flow cytometry using specific antibody or ligand binding approaches. RESULTS: After IFN-alpha (100 IU/ml) treatment cell surface EGFR expression was upregulated in 6 of 11 and down-regulated in 2 of 11 bladder cancer cell lines. The over expression of cell surface EGFR peaked within 48 to 96 hours and increased by 35% to 241% in individual cell lines. High level cell surface EGFR correlated with intracellular EGFR expression. Cell growth inhibition by IFN-alpha coexisted with EGFR over expression in the 6 lines. IFN-alpha treated cells remained sensitive to EGF treatment. CONCLUSIONS: IFN-alpha transiently up-regulates EGFR expression and inhibits in vitro growth in some human bladder cancer cells. IFN-alpha does not prevent EGFR from binding EGF or signal transduction via the EGF-EGFR pathway. This may have clinical implications for improving treatment based on EGFR targeting in select patients with bladder cancer.     

表皮细胞生长因子及其受体在胎儿和成人肠道组织中的表达特征

目的探讨表皮细胞生长因子(EGF)及其受体(EGFR)在不同发育阶段人小肠组织中的表达特征及其可能的生物学意义。方法24例被测标本中包括胎儿(胎龄13～31周)的小肠组织18例，成人(16～54岁)的小肠组织6例。用病理学技术和免疫组织化学方法确定EGF及EGFR两种蛋白在胎儿、成人小肠组织中的定位以及表达量的变化规律．结果EGF及EGFR在不同胎龄的胎儿和成人小肠组织中均有阳性表达。随着胎儿生长发育，小肠组织中EGF和EGFR的蛋白含量逐渐增加，这一变化趋势一直保持到成人小肠组织中。其中EGF主要存在于小肠黏膜上皮细胞、黏膜下层的血管内皮细胞内和浆膜上皮细胞的胞浆和胞外基质中，EGFR则分布于这些细胞的细胞膜上。结论EGF及EGFR在不同发育阶段的小肠组织中呈阳性表达，这种细胞因子可能以自分泌或旁分泌方式调节人肠道的生长发育、结构和功能的维持，并可能在小肠损伤后的修复中起重要作用。     

不同剂量己烯雌酚对新生小鼠睾丸表皮生长因子及受体的影响

目的 观察不同剂量己烯雌酚(DES)对新生小鼠睾丸组织中表皮生长因子(EGF)及其受体(EGFR)的影响,探讨DES对睾丸发育影响的机制.方法 建立小鼠DES模型.将72只怀孕的雌性昆明小鼠随机分成3组:正常组、对照组及实验组1～4(DES 10、25、50、100 μg/kg).采用免疫组织化学方法检测各组新生小鼠睾丸组织中EGF、EGFR的表达.结果 EGF和EGFR主要表达在新生小鼠睾丸的间质细胞.实验组1～4中EGF阳性细胞的累积吸光度值(IA)分别为75.43±1. 42、52.22±5.67、13.75±3.14、6.38±3.20,显著低于正常组及对照组中阳性细胞的IA值433.88±11.64、23.44±4.70;实验组EGFR阳性细胞的IA值分别为198.16±34.35、138.00±12.04、46.03±6.74、27.22±5.52,显著低于正常组及对照组中阳性细胞的IA值804.74±22.52、800.03±21.96.两者在正常、不同剂量DES实验组的两两比较差异有统计学意义(P＜0.05).随DES剂量增加,EGF和EGFR表达减弱,各组间差异有统计学意义(P＜0.05).EGF与EGFR呈强正线性相关(r=0.750,P＜0.01).结论 不同剂量DES对新生小鼠睾丸组织中EGF和EGFR表达强度均有影响,可能是影响睾丸发育机制之一.

Abstract：

Objective To investigate the effects of prenatal exposure to diethylstilbestrol (DES)with different dosages on epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in offspring mice testis, and the possible mechanism. Methods DES model was induced in mice by DES.Female Kungmiag mice were randomly divided into normal group, control group and experimental groups 1-4 ( DES 10, 25, 50, 100 μg/kg). Immunohistochemistry was applied to detect the expression of EGF and EGFR in the testicular tissue in each group. Results EGF and EGFR were expressed mainly in sffspring mice testis Leydig cells. The cumulative absorbance ( IA ) values of EGF positive cells in experimental groups 1-4 were 75. 43 ± 1.42, 52. 22 ± 5.67, 13.75 ± 3. 14, and 6. 38 ± 3.20 respectively, which were significantly lower than in normal and control groups (433.88 ± 11.64,423.44 ±4. 70 respectively).The IA values of EGFR positive cells in experimental groups 1-4 were 198. 16 ± 34. 35, 138.00 ± 12.04,46.03 ± 6. 74, 27.22 ± 5.52 respectively, which were significantly lower than in normal and control groups (804. 74 ± 22. 52,800. 03 ± 21.96 respectively). The expression levels of EGF and EGFR could be detected in each subgroup and statistically significant differences existed in the expression of EGF and EGFR between any two groups ( P ＜ 0. 05 ). With the increase of DES dosage, the expression of EGF and EGFR was decreased, with the difference being significant amond the groups ( P ＜ 0. 05 ). Conclusion DES can influence the expression of EGF and EGFR in mice testis, which might be one of possible mechanisms effecting the development of testis.     

EPIDERMAL GROWTH FACTOR BINDING BY MEMBRANES OF HUMAN RENAL CELL CARCINOMAS: ESTABLISHMENT OF AN EPIDERMAL GROWTH FACTOR RECEPTOR ASSAY FOR CLINICAL USE

In order to quantitate epidermal growth factor receptors (EGFR) in human renal cell carcinoma (RCC) tissues, the binding of isotope-labeled epidermal growth factor (¹²⁵I-EGF) to pooled membrane samples from human RCC was studied. Specific EGF binding to membranes at 4C reached a plateau after 2 h of incubation and remained constant up to 8 h; specific binding at 25C reached a plateau after 30 min of incubation, then decreased gradually. ¹²⁵I-EGF binding to the membrane was displaced by both EGF and transforming growth factor-aL, but not by insulin and basic fibroblast growth factor. Scatchard analysis of the specific EGF binding generated a straight line, indicating a single class of binding sites for EGF, with a dissociation constant (Kd) of 21.1 times10¹⁰M and a maximum number of binding sites of 57.2fmol/mg membrane protein. When the protein concentration in the incubation medium was adjusted from 0.71mg/ml to 2.84 mg/ml, Scatchard analysis revealed identical Kd values, and the maximum number of binding sites was proportional to the protein concentration. These results demonstrate the presence of EGFR on RCC membranes and indicate that these receptors can be studied quantitatively.     

Expression of human epidermal growth factor and its receptor in esophageal cancer

The expression of human epidermal growth factor (hEGF) was examined immunohistochemically in 86 esophageal cancer lesions, comprising 67 primary tumors and 19 metastatic lymph nodes. In the normal esophagus, the parabasal and intermediate cell layers showed a weak expression of hEGF, however, hEGF-positive tumor cells were detected in 62 (92.5 per cent) of the 67 primary esophageal carcinomas and in 18 (94.7 per cent) of the 19 metastatic lymph nodes. In this study, the immunoreactivity of hEGF was classified into 4 grades according to the number of stained tumor cells. A significant correlation was observed between the histologic type and the grade of hEGF immunoreactivity (Chisquare test, p<0.01). hEGF immunoreactivity in well differentiated squamous cell carcinomas was significantly higher than in other squamous cell carcinomas, although there were no correlations between other pathological findings and hEGF immunoreactivity. Patients with hEGF immunoreactivities of grades II or III had much worse prognoses than those with grades 0 or I (p<0.05). In 22 esophageal carcinomas and 10 normal esophageal mucosae, EGF receptor (EGFR) contents were measured by the competitive binding assay. The average EGFR content (101.3±35.7 fmol/mg protein, mean±SE) of the esophageal carcinomas was significantly higher than that (5.3±1.2) of the normal esophageal mucosae (p<0.05). Moreover, in hEGF negative tumors, EGFR contents were lower than in hEGF positive tumors. These results suggest that hEGF and EGFR show increased production in squamous cell carcinomas and could to be useful prognostic factors in patients with esophageal cancer.     

Survivin在膀胱移行细胞癌中的表达及其临床意义

目的比较Survivin在正常组织与膀胱移行细胞癌中表达的差异，分析其表达与膀胱移行细胞癌生物学特性及预后的关系。方法选用中山大学附属第三医院2001年至2006年间外科切除及活检的膀胱癌石蜡块标本40例，其中24例为浅表性膀胱癌组织，16例为浸润性膀胱癌组织。另取10例正常膀胱组织石蜡块标本，用免疫组化染色显示组织中Survivin表达，并对结果进行相应的统计学分析。结果10例正常膀胱组织中未见Survivin表达。于G，级、G：级、G，级膀胱癌组织中Survivin的表达率分别为20％（2／10）、38．9％（7／18）、75％（9／12）。经多因素分析，Survivin的表达与膀胱癌细胞的病理分级及患者的无瘤生存时间相关，而与其肿瘤分期及生存率不相关。结论正常膀胱组织不表达Survivin。Survivin的表达率随肿瘤细胞分化程度的降低而升高。Survivin的表达水平反映患者的无瘤生存率，是预测膀胱癌复发的独立预后因子。