脆性三联组氨酸基因在胃癌中的表达及其临床意义

目的:探讨FHIT基因在胃癌中的表达及意义。方法:采用免疫组化法检测49例胃癌、21例正常胃黏膜、31例萎缩性胃炎、27例异形增生组织中FHIT基因表达,并分析与临床分期、分化程度、淋巴转移及3年生存期的关系。结果:FHIT基因在进展期胃癌中的阳性表达率为18.4%,早期胃癌中为27.3%(3/11),均低于异型增生(55.6%,15/27)、萎缩性胃炎(74.2%,24/31)及正常胃黏膜(100%,P=0.000)中的表达。FHIT基因在早期胃癌中的表达高于中、晚期胃癌(P=0.001);在高分化组中低于中、低分化组(P=0.022),无淋巴结转移者与淋巴结转移者表达无统计学差别(P=0.065)。随访的49例胃癌资料中,FHIT基因表达阴性者生存时间明显低于表达阳性者(Log rank=5.06,P=0.0245)。结论:FHIT基因的表达缺失与胃癌的发生、发展及预后有关,FHIT水平可作为判断胃癌预后的一种指标。     

胃癌中脆性组氨酸三联体基因的甲基化和表达及其意义

目的检测脆性组氨酸三联体（FHIT）基因在胃癌中的蛋白表达及其甲基化情况，探讨FHIT在胃癌中的作用。方法收集胃癌新鲜组织标本共33例，采用免疫组织化学和甲基化特异性聚合酶链反应（PCR）的方法检测FHIT基因的蛋白表达和异常甲基化。结果33例胃癌组织中，24例表达FHIT蛋白，阳性率为73％，有13例发生了甲基化。阳性率为39％。胃癌中FHIT基因的蛋白表达和异常甲基化呈负相关。结论FHIT基因的异常甲基化是引起FHIT基因失活并导致胃癌发生的重要机制。     

乳腺癌中FHIT基因缺失与人乳头状瘤病毒感染相关性的研究

目的探讨乳腺癌组织中FHIT基因缺失的临床意义和人乳头状瘤病毒 (humanpapillo mavirus ,HPV)感染的关系。方法采用RT PCR检测了 38例乳腺癌及 10例正常乳腺组织中FHIT基因缺失情况 ,并用PCR技术检测HPV DNA片段。结果乳腺癌组织中FHIT基因缺失率及HPV DNA片段检出率分别为 6 3 1% ( 2 4/38)、31 6 % ( 12 /38) ,正常组织未检出FHIT基因缺失 ( χ2 =12 6 32 ,P <0 0 1;χ2 =4 2 11,P <0 0 5 )。FHIT基因缺失与乳腺癌组织分化程度、临床分期、淋巴结转移、远处转移密切相关 ;与 3年生存率呈负相关。在 12例HPV阳性组织中有 11例 ( 91 7% )发生FHIT基因缺失 ( χ2 =6 12 6 ,P <0 0 5 )。结论FHIT基因的缺失与乳腺癌发生发展密切相关 ,可作为判断乳腺癌恶性程度及预后的指标。HPV的感染与乳腺癌的发生有关。     

脆性组氨酸三联体基因蛋白在直肠癌组织中的表达及其与细胞凋亡的相关性研究

目的探讨脆性组氨酸三联体基因蛋白(FHIT)在直肠癌组织中的表达情况及其与细胞凋亡间的关系。方法利用组织芯片和免疫组织化学技术,检测16例正常直肠和16例腺瘤组织标本及80例直肠癌组织中的FHIT、Bax、Bcl-2和survivin基因蛋白的表达,并运用脱氧核糖核苷酸末端转移酶介导的缺口末端标记技术(TUNEL)检测直肠癌细胞凋亡。结果(1)直肠癌组织FHIT蛋白异常表达率为53.8%(43/80),其表达减弱与患者年龄、性别及肿瘤组织分型无关(P>0.05);而与肿瘤Dukes分期、淋巴结转移和患者的5年生存率关系密切(P<0.05,P<0.01)。(2)Bax、Bcl-2和survivin在直肠癌组织中的表达率分别为72.5%、51.3%和77.5%;survivin蛋白高表达与FHIT蛋白低表达密切相关(P<0.05);FHIT蛋白表达下降的同时,Bcl-2上调及Bax下调(P<0.01)。(3)FHIT蛋白表达减弱时细胞凋亡指数(AI)也下降;FHIT蛋白不同表达组间的AI比较,P<0.01。结论FHIT基因表达下降可能与直肠癌的发生密切相关,FHIT基因可能参与肿瘤细胞凋亡的调节。     

FHIT基因蛋白的表达与肝癌的临床关系

目的　探讨脆性组氨酸三联体 (FHIT)基因蛋白在原发性肝细胞癌 (简称肝癌 )中表达的临床意义。方法　采用免疫组织化学S -P法检测FHIT基因蛋白在 46例肝癌组织及癌旁组织和 10例正常肝组织中的表达。结果　FHIT基因蛋白在正常肝组织及癌旁组织均为阳性表达 ,显著高于在肝癌中的阳性表达率 5 6.5 2 %。FHIT基因蛋白的丢失与肿瘤是否有门静脉癌栓、肿瘤组织分化程度明显相关 (P <0 .0 5 ) ,与肿瘤大小、有无包膜、甲胎蛋白是否阳性、是否感染乙型肝炎病毒、是否合并肝硬化等因素均无明显的相关性。结论　FHIT基因蛋白缺失在肝癌组织中是频发事件 ,且FHIT基因蛋白的缺失和肝癌的临床表现及预后有一定的关联。     

基质金属蛋白酶-9和脆性组氨酸三联体基因在人类膀胱移行细胞癌中的表达及其意义

目的 探讨基质金属蛋白酶-9(MMP-9)和脆性组氨酸三联体基因(FHIT)在膀胱移行细胞癌组织中的表达、相互关系和临床意义.方法 采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)法观察50例膀胱移行细胞癌石蜡标本和10例正常膀胱组织中MMP-9和FHIT的表达.结果 50例膀胱癌组织中的FHIT和MMP-9的阳性表达率分别为58.0％和52.0％,与正常膀胱组织中的表达比较差异有统计学意义(P＜0.05和P ＜0.01).FHIT和MMP-9的表达与膀胱移行细胞癌的临床分期、病理分级和肿瘤的复发相关;肿瘤组织中的FHIT与MMP-9的表达呈负相关(r=-0.412,P＜0.01).结论 膀胱癌组织中FHIT和MMP-9的表达在肿瘤进展中发挥重要作用.     

肝细胞癌脆性组氨酸三联体Fhit蛋白表达丢失研究

目的　探讨肝细胞癌中FHIT基因蛋白 (Fhit)表达状况及其与临床病理指标的可能关系。方法　采用兔抗人Fhit蛋白抗体和枸橼酸 微波 SP免疫组化方法检测 83例福尔马林固定、石蜡包埋的肝细胞癌蜡块标本中Fhit表达状况并分析其与组织学分级、临床分期以及肿瘤大小的关系。结果　 83例肝细胞癌组织Fhit表达较正常肝组织明显降低或缺失者为 5 0例 (6 5 2 % ) ,与正常肝组织相等者为 33例 (34 8% )。Fhit蛋白低表达癌在癌组织学分级中的分布为Ⅰ级癌 4 6 7% (7/15 ) ,Ⅱ级癌 5 3 8% (2 1/ 39) ,Ⅲ级癌 75 9% (2 2 / 2 9) ,各级癌组间比较 ,相差无显著性 (P >0 0 5 )。Fhit蛋白低表达癌在临床分期中的分布为Ⅰ～Ⅱ期癌 2 1 6 % (8/ 37) ,Ⅲ～Ⅳ期癌 91 3% (42 / 4 6 ) ,二组比较 ,有非常显著性差异 (P <0 0 1)。Fhit蛋白低表达癌在非小肝癌 (>5 0mm)和 (≥ 30mm)病例组中的分布分别为 75 0 % (33/ 4 4)和 6 4 9% (46 / 71) ,而在≤ 5 0mm组和 <30mm(小肝癌 )组中则为4 3 6 % (17/ 39)和 33 3% (4/ 12 ) ,组间比较 ,均有显著性差异 (P均 <0 0 5 )。结论　肝细胞癌Fhit表达状况可能与临床分期以及肿瘤大小相关 ,提示Fhit表达降低可能对肝细胞癌的演化和进展具有重要作用并可能成为一个新的预后指标。     

FHIT在散发性结肠癌中的表达及意义

目的探讨散发性结肠癌中脆性组氨酸三联体(FHIT)基因蛋白表达状况及其与临床病理指标的可能关系。方法采用兔抗人FHIT蛋白抗体和枸橼酸微波SP免疫组织化学方法检测74例结肠癌26例正常组织、26例良性腺瘤标本中FHIT表达状况并分析其与Dukes’分期等临床病理指标以及5年生存率的关系。结果26例正常结肠组织中FHIT基因表达,1例阴性(1/26,3.8%)26例结肠腺瘤组织阴性7例(7/26,26.9%)。74例结肠癌组织中阴性35例(47.3%)。三者比较有显著性差异(P<0.05)。FHIT蛋白低表达与结肠癌患者的年龄、性别、肿瘤部位、大小、血清CEA的水平、组织学类型均无相关性(P>0.05);与分化程度、病理分期、淋巴转移、远处转移、五年生存率均有差异(P<0.05)。结论FHIT表达降低可能对结肠癌的演化和进展具有一定重要作用,并可能成为一个新的预后指标。     

Retinoblastoma gene mutations in primary human prostate cancer

Structural alterations in the entire coding regions (exons 1 to 27) of the retinoblastoma (RB) gene in primary human prostate cancers were investigated, using polymerase chain reaction and single strand conformational polymorphism analysis of RNA. Of 25 samples obtained from patients, four (16.4%) were found to have RB alterations. DNA sequencing of the PCR products revealed point mutations resulting in single amino-acid substitutions of exons 6 and 19 in two cases, and base deletions of exons 8 and 17 in two cases. Two of four cases with RB mutations were moderately differentiated localized tumors and other two with RB mutations were poorly differentiated tumors with metastases. Our results suggest that RB gene mutation is involved in progression steps of prostate carcinogenesis. © 1995 Wiley-Liss, Inc.     

Expression profile of an androgen regulated prostate specific homeobox gene NKX3.1 in primary prostate cancer

PURPOSE: NKX3.1, a member of the family of homeobox genes, exhibits prostate tissue specific expression and appears to play a role in mouse prostate development. Rapid induction of NKX3.1 gene expression in response to androgens has also been described. On the basis of the established role of androgens in prostatic growth and differentiation and studies showing an association of aberrant homeobox gene expression with the neoplastic process, we hypothesize that alterations of NKX3.1 gene expression play a role in prostate tumorigenesis. MATERIALS AND METHODS: NKX3.1 expression was analyzed in matched, microdissected normal and tumor tissues from 52 primary prostate cancer specimens from radical prostatectomy by semiquantitative RT-PCR and in situ hybridization and correlated with the clinicopathologic features. NKX3.1 expression was quantified as differential expression between matched tumor and normal tissues and was grouped as overexpression in tumor tissue, reduced expression in tumor tissue and no change between tumor and normal tissues. Androgen regulation of NKX3.1 expression was also studied in LNCaP cells. Androgen receptor (AR) expression in prostate tumor and normal tissue was correlated with NKX3.1 expression. RESULTS: Comparison of NKX3.1 expression between normal and tumor tissues revealed overexpression in 31% tumor specimens (16 of 52), decreased expression in 21% tumor specimens (11 of 52) and no change in 48% specimens (25 of 52). When these expression patterns were stratified by organ confined and non-organ-confined tumor, a higher percentage of patients exhibited NKX3.1 overexpression in non-organ confined tumor (40%) versus organ confined tumor (22%). Elevated NKX3.1 expression significantly correlated with tumor volume and serum prostate specific antigen (PSA) level in the NKX3.1 overexpression group (p<0.05). Metastatic prostate cancer cell lines did not exhibit mutations in the protein coding sequence of NKX3.1. Additionally, the NKX3.1 expression correlated with AR expression (p<0.01) in vivo in human prostate tissues. Comparison of PSA and NKX3.1 expression in response to androgen revealed a rapid androgen mediated induction of NKX3.1 expression in LNCaP cells. In situ hybridization analysis of representative specimens confirmed RT-PCR observations. CONCLUSIONS: These results suggest an association of NKX3.1 with a more aggressive phenotype of carcinoma of the prostate. Correlation of AR expression with NKX3.1 in human prostate tissues underscores the androgen regulation of NKX3.1 in the physiologic context of human prostate tissues.     

Casodex treatment induces hypoxia-related gene expression in the LNCaP prostate cancer progression model

Background

The changes in gene expression profile as prostate cancer progresses from an androgen-dependent disease to an androgen-independent disease are still largely unknown.

Methods

We examined the gene expression profile in the LNCaP prostate cancer progression model during chronic treatment with Casodex using cDNA microarrays consisting of 2305 randomly chosen genes.

Results

Our studies revealed a representative collection of genes whose expression was differentially regulated in LNCaP cells upon treatment with Casodex. A set of 15 genes were shown to be highly expressed in Casodex-treated LNCaP cells compared to the reference sample. This set of highly expressed genes represents a signature collection unique to prostate cancer since their expression was significantly greater than that of the collective pool of ten cancer cell lines of the reference sample. The highly expressed signature collection included the hypoxia-related genes membrane metallo-endopeptidase (MME), cyclin G2, and Bcl2/adenovirus E1B 19 kDa (BNIP3). Given the roles of these genes in angiogenesis, cell cycle regulation, and apoptosis, we further analyzed their expression and concluded that these genes may be involved in the molecular changes that lead to androgen-independence in prostate cancer.

Conclusion

Our data indicate that one of the mechanisms of Casodex action in prostate cancer cells is induction of hypoxic gene expression.     

脆性组氨酸三联体基因对结肠癌细胞增殖和凋亡的影响

目的 探讨脆性组氨酸三联体基因（FHIT）转染对结肠癌细胞株SW480增殖和凋亡的影响及其作用机制.方法 将重组真核表达质粒pRc/CMV2-FHIT通过脂质体转染技术导入人结肠癌细胞株SW480（实验组）,筛选稳定转染的细胞并扩增培养,以转染了空质粒pRc/CMV2的SW480细胞作为阴性对照,以正常SW480细胞作为空白对照.应用MTT法检测细胞的增殖活性,流式细胞仪检测细胞周期分布和细胞凋亡率,Western blot分析caspase-8酶原的变化,半定量RT-PCR检测caspase-8 mRNA水平的改变,肽核酸标记底物的比色法检测caspase-8的相对活性.结果 转染96 h后,实验组和阴性对照组细胞生长抑制率分别为71.7%和16.9%,G0/G1细胞比例分别为（63.3±3.5）%和（50.6±2.1）%,细胞凋亡率分别为（40.5±3.1）%和（18.6±2.6）%,caspase-8 mRNA条带光密度积分值分别为107和41,caspase-8蛋白相对活性分别为0.43和0.25;上述差异均有统计学意义（P＜0.05）.当加入FHIT抑制剂后,caspase-8蛋白相对活性恢复至对照组水平（0.22）.结论 FHIT基因转染能够明显抑制人结肠癌细胞株SW480的增殖,诱导SW480细胞发生G0/G1期阻滞,其作用机制可能与caspase-8表达及活性上调有关.     

Expression of a model gene in prostate cancer cells lentivirally transduced in vitro and in vivo

In a preclinical model for prostate cancer gene therapy, we have tested lentiviral vectors as a practical possibility for the transfer and long-term expression of the EGFP gene both in vitro and in vivo. The human prostate cancer cell lines DU145 and PC3 were transduced using experimental conditions which permitted analysis of the expression from a single proviral vector per cell. The transduced cells stably expressed the EGFP transgene for 4 months. After injection of the transduced cell populations into Nod-SCID mice a decrease in EGFP was only observed in a minority of cases, while the majority of tumors maintained transgene expression at in vitro levels. In vivo injection of viral vector preparations directly into pre-established subcutaneous or orthotopic tumor masses, obtained by implantation of untransduced PC3 and DU145 cells led to a high transduction efficiency. While the efficiency of direct intratumoral transduction was proportional to the dose of virus injected, the results indicated some technical limitations inherent in these approaches to prostate cancer gene therapy.     

脆性组氨酸三联基因在肿瘤相关研究中的状况

脆性组氨酸三联基因(ragile histidine triad,FHIT)是近年来克隆出的一个新候选抑癌基因，定位于染色体3p14．2，其cDNA长约1．1kb，含10个外显子。FHIT基因包含脆性部位FRA3B和t(3；8)易位断裂处，编码的蛋白是一种典型的Ap3A水解酶。FHIT基因在人类多种肿瘤中均有不同类型的高比例缺失。目前的研究显示FHIT的缺失和人类众多肿瘤的发生发展相关。     

Microarray analysis of differential gene expression in androgen independent prostate cancer using a metastatic human prostate cancer cell line model

Progression to androgen independence (AI) leading to uncontrolled cell growth is the main cause of death in prostate cancer. While almost all patients with metastatic prostate cancer will initially respond to anti-androgen treatments, the majority will fail hormonal treatments in less than 2 yrs. Both genetic and epigenetic alterations in gene expression contribute significantly to the development of AI. To investigate this we have used an in vitro cell line model of AI prostate cancer from which we have identified a number of differentially expressed genes associated with progression to AI in prostate cancer. We used an in vitro cell line model of AI prostate cancer, to study differential gene expression using cDNA microarray analysis and corroborated the microarray results with Ribonuclease Protection Assay (RPA). Approximately 4480 out of 7075 (63.3%) cDNA cloned genes were differentially expressed, of which, 6 genes were differentially expressed by at least fivefold. RPA was used to corroborate the microarray results for the five most highly differentially expressed genes. Using an in vitro cell line model and microarray analysis we have identified a number of candidate genes for further investigation in AI prostate cancer.     

脆性组氨酸三体和血管内皮生长因子在前列腺癌组织中的表达

目的探讨脆性组氨酸三体（FHIT）与血管内皮生长因子（VEGF）在前列腺癌中的表达及意义。方法选取不同分级的前列腺癌组织和前列腺增生组织,采用免疫组织化学SP法染色,检测FHIT及VEGF的表达情况。结果64例前列腺癌组织中病理分级（按Gleason分级系统）分化良好癌（Gleason2～4分）、中等分化癌（Gleason5～7分）、分化不良癌（Gleason8～10分）中FHIT的蛋白表达率分别为84．2％（16／19）、45．4％（10／22）、17．4Y00（4／23）。随着Gleason评分增加,FHIT蛋白表达率显著下降（P〈0．05）。病理分级中分化良好癌、中等分化癌、分化不良癌VEGF的蛋白表达率分别为42．0％（8／19）、63．6％（14／22）、86．9％（20／23）,随着Gleason评分增加,VEGF蛋白表达率显著升高（P〈0．05）。结论FHIT的表达与前列腺癌的Gleason评分、VEGF的表达负相关;VEGF的表达与Gleason评分正相关。     

Mechanical strain alters gene expression in an in vitro model of hypertrophic scarring

Fibroblasts represent a highly mechanoresponsive cell type known to play key roles in normal and pathologic processes such as wound healing, joint contracture, and hypertrophic scarring. In this study, we used a novel fibroblast-populated collagen lattice (FPCL) isometric tension model, allowing us to apply graded biaxial loads to dermal fibroblasts in a 3-dimensional matrix. Cell morphology demonstrated dose-dependent transition from round cells lacking stress fibers in nonloaded lattices to a broad, elongated morphology with prominent actin stress fibers in 800-mg-loaded lattices. Using quantitative real-time RT-PCR, a dose dependent induction of both collagen-1 and collagen-3 mRNA up to 2.8- and 3-fold, respectively, as well as a 2.5-fold induction of MMP-1 (collagenase) over unloaded FPCLs was observed. Quantitative expression of the proapoptotic gene Bax was down-regulated over 4-fold in mechanically strained FPCLs. These results suggest that mechanical strain up-regulates matrix remodeling genes and down-regulates normal cellular apoptosis, resulting in more cells, each of which produces more matrix. This "double burden" may underlie the pathophysiology of hypertrophic scars and other fibrotic processes in vivo.