FHIT基因在结直肠癌中的表达及其与细胞凋亡抑制的相关性

Objective： To investigate the expression of fragile histidine triad （FHIT） protein in normal colorectal tissue, colorectal adenoma and colorectal cancer （CRC）, and to study the relationships between the expression of FHIT protein and the clinical pathology, the apoptosis-associated protein （Bcl-2, Bax, Survivin）, apoptosis in colorectal cancer. Methods： Tissue microarray （TMA） and immunohistochemistry SP were used to detect the expression of FHIT gene, Bcl-2, Bax and Survivin in 16 cases of the normal colorectal tissue, 16 cases of colorectal adenoma and 80 cases of the colorectal cancer. TUNEL was used to detect the apoptosis index （Al） in 80 cases of the colorectal cancer. Results： （1） The positive rates of FHIT gene expression in normal colorectal tissue, colorectal adenoma and adenocancer were 93.75%, 68.75% and 46.25% respectively. There were no significant differences in the relationships between the FHIT gene expression and histological types, the gender as well as the age （P〉0.05）. There were significant relationships between FHIT gene expression and lymph node metastasis, histological grades, Duke＇s system as well as the 5-year survival rate after operation. （2） The positive rates of Bax, Bcl-2 and Survivin in colorectal adenocancer were 72.50%, 51.25%, 77.50% respectively. The expression of FHIT gene was positively correlated with that of Bcl-2, Bax and Survivin. （3） The mean AI in FHIT negative tumors was significantly lower than that in FHIT positive tumors （P〈0.01）. Conclusion： FHIT gene may play a role in the oncogenesis and progression of colorectal cancer. The abnormal regulation of apoptosis may play an important role in the pathogenesis of colorectal cancer.     

Expression of Hpa and CD222 in bladder carcinoma and analysis of clinico-pathologic correlation

Objective： Our study aimed to investigate the relationships between the clinico-pathologic features and the heparanase （Hpa） and CD222 expressions in bladder carcinoma. Methods： The expressions of Hpa and CD222 in 95 bladder carcinoma specimens and 20 paraneoplastic bladder tissues （controls） were assessed using the immunohistochemical stain- ing method. Results： The positive expression rates of Hpa and CD222 in bladder carcinoma were 68.42% and 61.05%, respectively. The positive rate of Hpa was significantly higher in the carcinoma specimens than in the control specimens （P 〈 0.01）. Similarly, the Hpa expression in the invasive bladder carcinoma was significantly higher than that in the non-invasive bladder carcinoma （P 〈 0.01）. A positive correlation was observed between the expressions of Hpa and CD222 （P 〈 0.05）. The expressions of Hpa and CD222 were significantly correlated with lymphatic invasion and TNM staging （P 〈 0.05）. The 5-year survival rate was significantly higher in negative expression of the Hpa group than that in the positive expression group （P 〈 0.05）. Compared with the non-co-positive expression group, the 5-year survival rate in the co-positive expression of Hpa and CD222 group was significantly lower （P 〈 0.05）. Conclusion： High Hpa and CD222 expressions in tumor tissues were associated with the occurrence and development of bladder carcinoma. Our results provide helpful information for the further diagnosis and therapy of bladder carcinoma.     

乳腺良恶性疾病端粒酶hTERT及调亡相关蛋白的表达

Objective： To analyse the expression of telomerase and apoptosis related protein, and explore the possible mechanism of breast cancer development. Methods： Immunohistochemistry method （SP） was used to detect the expression of hTERT, p53 and bcl-2 in the tissues of 48 cases of human breast cancer and 42 cases of benign lesions in breast. Results： The positive rates of expression of hTERT, p53 and bcl-2 in breast cancer were 87.50%, 56.25% and 54.17%, respectively. Compared with the groups of adjacent noncancerous and benign lesions, there was a significant difference among three types of tissues （P 〈 0.05）. The positive rates of expression of p53 and bcl-2 in the group with positive expression of hTERT were 64.28% and 61.90%, respectively, and their difference was significant compared with the negative group （P 〈 0.05）. Conclusion： There is a correlation between the activation of telomerases and p53 gene mutation in the development of breast cancer, and they are perhaps relation to the down regulation of bcl-2.     

Relationship between FHIT expression and HPV status in vulvar squamous cell carcinomas

OBJECTIVE To investigate the relationship between fragile histidine triad （FHIT） depletion and human papillomavirus （HPV） status in the vulvar squamous cell carcinomas （VSCC）. METHODS Immunohistochemical method was used to detect the expression of FHIT protein in 42 cases of VSCC and 10 cases of normal vulvar tissues. PCR was used to detect HPV infection status. We analyzed the relationship of the expression of FHIT protein between the 2 groups： HPV positive and HPV negative, as well as the clinically pathological characteristics. RESULTS The expression of FHIT was positive in all normal vulvar tissues, and 71.4% was depletion in VSCC （P 〈 0.01）. Abnormal expression of FHIT was significantly correlated with pathological grade （P 〈 0.05）. There was a significant difference between FHIT depletion and HPV infection based on the statistial analysis （P 〈 0.05）. CONCLUSION The FHIT depletion is related to occurrence and development of VSCC, and the abnormal expression of FHIT significantly correlates with HPV infection.     

HYPERMETHYLATION OF p14^ARF PROMOTER REGION AND EXPRESION OF p14^ARF GENE PRODUCT IN NON-SMALL CELL LUNG CANCER

Objective： This study was designed to investigate promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer, and value the role of p14^ARF promoter methylation in carcinogenesis of non-small cell lung cancer. Methods： Promoter methylation status and protein expression of p14^ARF gene in 40 cases of non-small cell lung cancer were analyzed by methylation specific polymerase china reaction （MSP）, restriction enzyme-related polymerase chain reaction （RE-PCR） and immunohistochemistry （IHC）. Results： The positive rates of p14^ARF promoter methylation in tumor tissues and normal tissues adjacent to cancer were 17.5% （7/40） and 2.5% （1/40） respectively. There were statistically significant differences between them, P〈0.05. The results of RE-PCR were consistent with that of MSP. The expression rate of p14^ARF protein in tumor tissues was significantly lower than that in normal tissues adjacent to cancer, p〈0.01. Promoter methylation status and protein expression of p14^ARF gene in non-small cell lung cancer showed significantly an inverse correlation （r=-0.56, P〈0.01）, and both of them did not relate statistically with the clinicopathologic characteristics of patients such as histological classification, clinical stage, differentiation grade and lymph node involvement. Conclusion： Promoter methylation is a crucial mechanism of inactivation of p14^ARF gene. Promoter methylation of p14^ARF gene might he involved in carcinogenesis of non-small cell lung cancer, and is an early event in development process of non-small cell lung cancer. It might be used as a new target in gene treatments in the future.     

Expression of Cyclin E and Its Relationship with the Prognosis of Patients with Breast Cancer

Objective： To investigate the expression of cyclin E in breast cancer tissues and its relationship with prognosis of the patients with breast cancer. Methods： The expression of cyclin E, HER-2/neu, nm23-H1 and actin was detected in 80 breast cancer tissues and 18 benign breast tumor tissues by immunohistochemical methods. The relationship between cyclin E and the remaining genes or the clinical data of the patients with breast cancer was analyzed. Results： The over expression rate of cyclin E in malignant tissues was obviously higher than that in benign tumor tissues （P〈0.01）. The over expression of cyclin E in later stage of disease was higher than that in early stage of disease （P〈0.05）. The expression of cyclin E in ER positive tissues was lower than that in ER negative tissues （P〈0.05）. The expression of cyclin E in PR positive tissues and PR negative tissues had no significant difference （P〉0.05）. The expression of cyclin E in HER-2/neu positive tissues was higher than that in HER-2/neu negative tissues （P〈0.05）. And the expression of cyclin E in ER, PR and HER-2/neu all positive tissues was much higher （P〈0.01）. There was no significant difference in the expression of cyclin E between nm23-H1 positive tissues and nm23-H1 negative tissues （P〉0.05）. The expression of cyclin E in actin positive and continuous distribution tissues was lower than that in actin negative or discontinuous distribution tissues （P〈0.05）. Conclusion： The expression of cyclin E has a strong correlation to the prognosis of the patients with breast cancer.     

P27蛋白在人脑胶质瘤组织中的表达及其临床意义

Objective： To study p27/Kipl expression in gliomas and its application value. Methods： Imo munohistochemical technique was used to detect the exprssion of p27/Kipl gene in 48 different malignant grade human brain glioma tissues categorized according to WHO classification and 12 normal human brain tissues,which were analyzed quantitatively by using the image system and compared retrospectively with the patients＇ clinical characteristics. Results： In this series, the immunohistochemical reaction for p27/Kipl was confined to the nuclei. The abnormal positive expression rate of p27/Kipl in gliomas was found to be higher than that in normal tissues （P〈0.05）. The positive nuclei expression of p27/Kipl decreased in number and staining intensity with the increasing degree of histological malignancy （P〈0.05）. Lower expression of p27/Kipl was associated with poor prognosis （P〈0.05）. P27/Kipl expression could be regarded as an independent prognostic factor. Conclusion： The abnormal expression of p27/Kipl may be closely related to the occurrence and development of gliomas, and also can be used to evaluate the prognosis independently.     

Cx26基因修饰对人胃癌MGC-803细胞增殖和迁移能力的影响

目的 探讨Cx26基因对人胃癌MGC-803细胞的生长、细胞周期及迁移的影响.方法 将重组表达质粒pBudCE4.1-Cx26转染至人胃癌MGC-803细胞(转染组),另设空白组(未经任何特殊处理)和对照组(空载质粒组).收集40例胃癌组织及对应癌旁正常组织.通过RT-PCR和Western blot法测定细胞及组织中的Cx26 mRNA及蛋白表达水平.划痕染料示踪法检测细胞间通讯功能.采用MTF比色法及Transwell迁移实验分别检测转染后细胞增殖及迁移的变化,Western blot法检测各组细胞凋亡相关蛋白(Bax、Bcl-2、caspase 3)表达情况,流式细胞仪检测转染后细胞周期变化.结果 Cx26基因mRNA和蛋白在人胃癌组织中的表达量低于癌旁组织,差异均具有统计学意义(均P＜0.05).Cx26基因修饰人胃癌MGC-803细胞后,转染组与对照组和空白组比较,Cx26 mRNA和蛋白表达升高(均P＜0.05),细胞传递的荧光强度亦升高(P＜0.01).转染后48 h、72 h细胞增殖均受到抑制(均P＜0.01).转染24 h后,转染组MGC-803细胞穿膜数低于对照组和空白组(均P＜0.01),细胞凋亡相关蛋白Bax和caspase 3表达均增加(均P＜0.01),Bel-2表达各组间比较差异无统计学意义(P＞0.05).转染后48 h,转染组MGC-803细胞被阻滞在G2期(P＜0.01).结论 转染Cx26基因对人胃癌MGC-803细胞的增殖与迁移具有抑制作用,对细胞凋亡有促进作用,并可阻滞细胞在G2期.     

膀胱癌细胞Cx43对顺铂抗肿瘤作用影响研究

目的 连接蛋白43(connexin43,Cx43)在人体正常组织及肿瘤组织中广泛表达,并且参与细胞的生长控制和组织分化.本研究探讨采用RNA干扰技术沉默膀胱癌5637细胞株Cx43蛋白表达对顺铂化疗敏感性的影响.方法 体外培养膀胱癌5637细胞和正常尿路上皮SV-HUC-1细胞,采用蛋白质印迹法检测膀胱癌5637细胞系和正常尿路上皮SV-HUC-1细胞中Cx43蛋白表达,应用免疫荧光技术检测膀胱癌5637细胞系中Cx43蛋白的定位.采用RNA干扰技术沉默膀胱肿瘤5637细胞株Cx43蛋白的表达并用顺铂(3μg/mL)处理SiRNA-Cx43组(实验组)和SiRNA-Control(对照组)后,通过CCK-8检测实验组、对照组及野生型5637细胞的增殖情况;采用流式细胞术检测两组癌细胞的凋亡;采用蛋白质印迹法检测顺铂处理后野生型5637细胞后细胞中Cx43、Cleaved Caspase-3的表达及实验组和对照组中Cx43、Cleaved Caspase-3和Bcl-2的表达.结果 膀胱癌5637细胞中Cx43表达较正常尿路上皮细胞高,相对蛋白表达量分别为1.013±0.102和0.556±0.054,两细胞系比较差异有统计学意义,t=3.789,P=0.019;免疫荧光检测Cx43主要定位于细胞质中.CCK-8结果显示,膀胱癌5637细胞随着顺铂药物的浓度(0.75、1.5、3和6μg/mL)和时间(0、1、2、3 d)的增加,细胞的增殖减少,采用重复测量方差分析,各组间比较,差异有统计学意义,F=153.634,P＜0.001;实验组增殖较对照组明显减少,差异有统计学意义,F=9.949,P=0.02.流式细胞仪检测结果显示,实验组和对照组凋亡率分别为(63.00±4.58)％和(34.33±6.03)％,差异有统计学意义,t=7.457,P＜0.01.蛋白质印迹法结果显示,5637细胞随着药物(顺铂3 μg/mL)作用时间增加,Cx43蛋白表达逐渐降低,差异有统计学意义,F=178.868,P＜0.001;而CleavedCaspase-3逐渐升高,差异有统计学意义,F=21.643,P＜0.001.顺铂(3μg/mL,4h)处理实验组和对照组后,CleavedCaspase-3蛋白相对表达量分别为0.740±0.092和0.373±0.091,差异统计学意义,t=6.394,P=0.001;BclL-2蛋白相对表达量分别为0.260±0.066和0.817±0.068,差异统计学意义,t=4.814,P=0.005 结论 沉默膀胱癌5637细胞中Cx43蛋白表达能提高顺铂的敏感性,可能与反常定位于细胞质中的Cx43参与线粒体介导的凋亡途径有关.     

Caspase-3与Bcl-2蛋白在膀胱移行细胞癌中的表达及意义

背景和目的:肿瘤的发生是细胞异常增生和凋亡不足的结果。以往研究认为凋亡的发生与Caspase-3和Bcl-2蛋白有密切的联系。本研究拟通过检测Caspase-3和Bcl-2蛋白在膀胱移行细胞癌中的表达,探讨其在膀胱移行细胞癌发生、发展中的意义。方法:采用免疫组织化学(SP)法,对52例膀胱移行细胞癌组织和10例正常膀胱粘膜组织中的Caspase-3及Bcl-2蛋白表达情况进行检测。结果:膀胱移行细胞癌组织中的Caspase-3蛋白表达率(53.8%,28/52)明显低于正常膀胱粘膜组织(90.0%,9/10),Caspase-3的蛋白表达与膀胱移行细胞癌的病理分级有关(P<0.05),但与临床分期没有明显的联系;膀胱移行细胞癌组织中的Bcl-2蛋白表达率(51.9%,27/52)明显高于正常膀胱粘膜组织(20.0%,2/10),且Bcl-2蛋白表达情况与病理分级、临床分期均无关(P>0.05)。在正常膀胱粘膜中Caspase-3与Bcl-2的蛋白表达呈负相关(rs=-0.659,P<0.01)。结论:Caspase-3蛋白的高表达与Bcl-2蛋白的低表达在膀胱癌的发生和细胞凋亡的调控方面可能起着重要作用。     

E2F3 及C-myc 在膀胱移行细胞癌组织中的表达及其相关性研究*

目的:探讨E2F3 及C-myc 在膀胱癌组织中表达的相互关系及其与膀胱癌恶性生物学行为之间的关系。方法:应用免疫组织化学方法检测不同病理分期及组织分级的BTCC标本和正常膀胱黏膜中E2F3 及C-myc 蛋白的表达情况,应用免疫组化自动分析系统进行分析,结果以积分光密度表示。结果:膀胱移行细胞癌组织中E2F3 及C-myc 蛋白的表达阳性率明显高于正常膀胱黏膜,两组间有显著性差异（P<0.05）,同时E2F3 的表达率与膀胱癌的分级和分期密切相关（P<0.05;P<0.01）;C-myc 的表达率与膀胱癌的分级和分期密切相关（P<0.01;P<0.01）。 膀胱移行细胞癌中E2F3 与C-myc 的表达呈正相关。结论:E2F3 及C-myc 与膀胱癌的恶性程度密切相关,同时二者之间的表达密切相关,二者不仅可以作为膀胱癌的诊断与预后指标,而且为以E2F3 为靶点的膀胱癌的基因治疗提供理论依据。      

TMS1/ASC基因CpG岛甲基化与膀胱移行细胞癌发生的关系

目的检测抑癌基因TMS1/ASC启动子区5′CpG岛甲基化状态及其在膀胱移行细胞癌（BTCC）中mRNA和蛋白表达水平。方法应用MSP技术检测膀胱移行细胞癌中TMS1/ASC基因启动子区甲基化状态,RT-PCR和Western blot法分别检测其mRNA和蛋白表达水平。结果TMS1/ASC基因在正常膀胱组织中未发生甲基化,而在癌组织中甲基化频率为46.9%（15/32）,并且随着肿瘤分级、分期的增加,其甲基化水平逐渐升高（χ^2=23.106,P〈0.05）。在15例启动子异常甲基化的BTCC标本中,14例同时伴有TMS1/ASC基因表达缺失或下调,两者存在明显的相关性（γ=0.5842,P〈0.05）。TMS1/ASC mRNA和蛋白表达在正常膀胱组织和BTCC组织中分别为81.3%（26/32）、18.8%（6/32）（P〈0.01）,不同病理分级、临床分期间差异有统计学意义（P〈0.05）。结论TMS1/ASC基因启动子区异常甲基化可能导致该基因转录表达失活,使其mRNA和蛋白表达减少,甚至缺失,这可能是膀胱癌发生、发展的原因之一。     

Ki-67抗原及细胞周期素D1在初发和复发性膀胱移行细胞癌中表达的研究

 目的 探讨Ki-67抗原、细胞周期素D1（CyclinD1）在初发和复发性膀胱移行细胞癌组织中的表达及其临床意义。方法 收集47例膀胱移行细胞癌组织（膀胱移行细胞癌组），所有标本均经病理证实。肿瘤分期按UICC及TNM标准分为：浅表性肿瘤Tis ~ T1期14例，浸润性肿瘤T2 ~ T4期33例；肿瘤病理分级按WHO标准分为G1期21例，G2期22例，G3期4例；其中14例为复发；收集13份正常膀胱黏膜组织作对照组。采用免疫组织化学SP法检测组织中Ki-67抗原、CyclinD1的表达。结果 膀胱移行细胞癌组Ki-67抗原阳性率为87.23 ％，对照组为15.38 ％，两组比较差异有统计学意义（P＜0.001）；Ki-67抗原表达与肿瘤分级分期相关（P＜0.001）；复发病例Ki-67抗原表达明显高于初发病例（P＜0.05）。膀胱移行细胞癌组CyclinD1阳性率为91.48 ％，对照组为7.69 ％，差异有统计学意义（P＜0.001）；CyclinD1表达与肿瘤分级呈负相关（r =-0.384，P＜0.01），同分期、复发未见相关性。结论 Ki-67抗原、CyclinD1能较准确地评估膀胱移行细胞癌的生物学行为。     

Hp感染与胃癌组织中Bcl-2、Bax表达的相关性研究

目的：揭示Bcl-2,Bax在胃癌组织的表达与Hp感染的相关性。方法：采用快速尿素酶试验及组织学改良Giemsa染色法联合检测130例胃癌组织与70例慢性胃炎组织中Hp感染情况,免疫组织化学PV9000法,检测130例胃癌组织与70例慢性胃炎组织中Bcl-2,Bax蛋白的阳性表达情况。结果：Hp（＋）组Bcl-2蛋白的阳性表达率明显高于Hp（-）组（P〈0.01）;胃癌组织中Hp感染与Bcl-2蛋白表达之间存在正相关关系（P〈0.01,r=0.288）;Hp（＋）组Bax蛋白的阳性表达率明显低于Hp（-）组（P〈0.01）;胃癌组织中Hp感染与Bax蛋白表达之间存在负相关关系（P〈0.01,r=-0.536）。结论：Hp感染与Bcl-2蛋白表达存在正相关性,与Bax蛋白表达存在负相关性,提示Hp感染与细胞凋亡,二者可能共同参与胃癌的发生发展过程。     

乳腺癌组织中上皮钙黏蛋白和B细胞淋巴瘤基因2的表达及其临床意义的研究

目的:探讨乳腺癌中上皮钙黏蛋白（E-cadherin）和B细胞淋巴瘤基因2（Bcl-2）的表达及其临床意义。方法:使用免疫组织化学方法检测68例乳腺癌旁正常组织、41例纤维腺瘤组织和74例乳腺癌组织的E-cadherin和Bcl-2的表达水平,并分析乳腺癌患者各病理特征与E-cadherin和Bcl-2表达的相关性。结果:乳腺癌旁正常组织和纤维腺瘤组织中E-cadherin阳性率高于乳腺癌组织,Bcl-2阳性率低于乳腺癌组织,差异具有统计学意义（P均＜0.05）。乳腺癌组织病理分级越高、TNM分期越严重、有淋巴结转移和复发的患者 E-cadherin阳性率低,Bcl-2阳性率高;ER阳性患者E-cadherin阳性率高,Bcl-2阳性率高。组织病理分级低-中分化患者E-cadherin阳性率高于高分化患者,Bcl-2阳性率低于高分化患者,差异具有统计学意义（P均＜0.05）;TNM分期Ⅰ+Ⅱ期患者E-cadherin阳性率高于Ⅲ期,Bcl-2阳性率低于Ⅲ期,差异具有统计学意义（P均＜0.05）;有淋巴结转移患者E-cadherin阳性率低于无转移者,Bcl-2阳性率高于无转移者,差异具有统计学意义（P均＜0.05）;雌激素受体（ER）阴性患者E-cadherin、Bcl-2阳性率低于ER阳性者,差异具有统计学意义（P均＜0.05）;复发患者E-cadherin阳性率低于无复发者,Bcl-2阳性率高于无复发者,差异具有统计学意义（P均＜0.05）;Bcl-2的表达与E-cadherin的表达存在负相关性（r=-0.638,P＜0.05）。结论:乳腺癌组织中E-cadherin表达降低,Bcl-2表达升高,二者与乳腺癌组织病理分级、TNM分期、淋巴结转移、ER和复发有关,E-cadherin和Bcl-2的表达存在相关性。     

Hp感染与胃癌组织中Bcl-2、Bax表达的相关性研究

目的 揭示Bcl-2,Bax在胃癌组织的表达与Hp感染的相关性.方法 采用快速尿素酶试验及组织学改良Giemsa染色法联合检测130例胃癌组织与70例慢性胃炎组织中Hp感染情况,免疫组织化学PV9000法,检测130例胃癌组织与70例慢性胃炎组织中Bcl-2,Bax蛋白的阳性表达情况.结果 Hp(+)组Bcl-2蛋白的阳性表达率明显高于Hp(-)组(P<0.01);胃癌组织中Hp感染与Bcl-2蛋白表达之间存在正相关关系(P<0.01,r=0.288);Hp(+)组Bax蛋白的阳性表达率明显低于Hp(-)组(P<0.01);胃癌组织中Hp感染与Bax蛋白表达之间存在负相关关系(P<0.01,r=-0.536).结论 Hp感染与Bcl-2蛋白表达存在正相关性,与Bax蛋白表达存在负相关性,提示Hp感染与细胞凋亡,二者可能共同参与胃癌的发生发展过程.     

膀胱移行细胞癌组织中透明质酸合成酶基因家族差异表达的临床意义

目的 研究透明质酸合成酶(HAS)家族基因在膀胱移行细胞癌(BTCC)组织中的差异表达情况,并探讨其潜在的临床意义.方法 运用实时定量聚合酶链反应(PCR)方法,检测78例不同恶性程度的BTCC组织中各HAS亚型mRNA的表达.运用Western blot方法,检测78例BTCC组织和12例正常人膀胱黏膜中各HAS亚型的蛋白表达.分析BTCC组织中各HAS亚型mRNA和蛋白表达与临床病理特征的关系.结果 78例BTCC组织中,HAS2、HAS2和HAS3的mRNA中位表达水平分别为0.0123、0.0353和0.0717,三者差异有统计学意义(P＜0.01).12例正常膀胱黏膜组织未见任何HAS亚型的蛋白表达.78例BTCC组织中,HAS2、HAS2和HAS3的蛋白表达量分别为0.5685±0.2510、1.005±0.5159和0.5040±0.3604,其中HAS1和HAS2、HAS2和HAS3的蛋白表达差异有统计学意义(P=0.004,P=0.001),而HAS1和HAS3的蛋白表达差异无统计学意义(P=0.654).HAS1 mRNA的表达与肿瘤发病、T分期、病理分级有关,HAS2 mRNA的表达与肿瘤大小、肿瘤数量、肿瘤发病、T分期、病理分级有关,HAS3 mRNA的表达水平与各临床病理特征均无关.HAS1蛋白表达与肿瘤发病、T分期和病理分级有关,HAS2蛋白的表达与肿瘤大小有关,HAS3蛋白表达与各临床病理特征无关.结论 膀胱移行细胞癌组织中,3种HAS亚型均明显异常高表达,提示透明质酸作为一种尿源性膀胱癌标记物比较可靠.相对于HAS1和HAS3,HAS2在BTCC的发生、发展过程中可能具有更为重要的作用.

Abstract：

Objective To investigate the differential expression of the hyaluronic acid synthase (HAS) family in human bladder transitional cell carcinoma(BTCC) and its potential clinical significance. Methods The relative quantitative detection of the expression of HAS isoforms(HASs) was performed in 78 human BTCC tissues(mRNA & protein)and 12 normal human bladder mucosa(protein) by real-time RTPCR and Western blot,and the results were statistically analyzed according to the clinical data. Results All the BTCC tissues expressed three HAS isoform mRNA and protein,but to a different extent,as for mRNA,HAS3 ＞ HAS2 ＞ HAS1(P ＜0.001),with a significant difference in HAS1/HAS2,HAS1/HAS3 and HAS2/HAS3(P = 0.003,＜0.001,0.006,respectively).Among the proteins,the HAS2 expression was the highest,with a significant difference in HAS1/HAS2,and HAS2/HAS3(P = 0.004,0.001,respectively),but not in HAS1/HAS3.The elevation of HAS1 mRNA and protein expression was significantly related with the tumor malignancy,tumor initial onset/recurrence,T1/T2 and T1/T3-4 stags,and tumor grading(P=0.02,＜0.001,0.038,＜0.001;0.025,0.031,0.023,0.002;respectively).The HAS2 mRNA expression was significantly related with tumor size(diameter ≤ 3.0 cm/＞ 3.0 cm),tumor number(single or multiple),tumor initial onset/recurrence,T-staging,and histopathological differentiation(low grade/high grade)(P=0.012,0.004,＜0.001,＜0.001,＜0.001,respectively),but its protein expression was not significantly different in all subgroups except with the tumor size(mass diameter ＞3.0 cm/≤3.0 cm).However,HAS3 mRNA and protein expression had no significant difference among all the subgroups.In normal human bladder mucosa,no HAS expressions were detected. Conclusions The abnormally high expression of the HASs further indicate the reliability of hyaluronan as a urinary marker for human BTCC.Compared with HAS1 and HAS3,HAS2 as a marker may have more usefulness in studies on human BTCC carcinogenesis or development.The high expression of HAS1 protein seems to play a more important role in the BTCC tumorigenesis,and may indicate a poor prognosis of the BTCC patients.