肾细胞癌中抑癌基因PTEN的表达及生物学意义

目的 :研究抑癌基因PTEN在肾细胞癌的表达及其生物学意义。方法 :应用免疫组织化学S P法检测 5例正常肾组织、18例癌旁肾组织和 40例肾细胞癌组织中抑癌基因PTEN的表达。结果 :5例正常肾组织和 18例癌旁肾组织均有较强的PTEN蛋白的表达 ,二者PTEN蛋白的表达强度、阳性细胞的分布形式无差异。抑癌基因PTEN在肾细胞癌中的表达不同于正常肾组织和癌旁肾组织 ,12 5 %的肾细胞癌呈PTEN蛋白阴性 ;17 5 %的肾细胞癌PTEN蛋白呈弱阳性 ;70 %的肾细胞癌PTEN蛋白呈阳性或强阳性 ,与癌旁组织PTEN蛋白的染色强度无差异。PTEN蛋白阴性的肾细胞癌 ,肾门淋巴结转移率为80 % ;PTEN蛋白阳性的肾细胞癌 ,肾门淋巴结转移率为 2 0 %。PTEN蛋白阴性肾细胞癌的肾门淋巴结转移率与PTEN阳性肾细胞癌的肾门淋巴结转移率比较 ,差异有显著性 (P <0 0 5 )。结论 :肾细胞癌中存在着抑癌基因PTEN的表达缺失和异常 ;抑癌基因PTEN的表达异常可能与肾细胞癌的发生、发展有关 ,抑癌基因PTEN是肾细胞癌的一种新的相关基因。     

肾损伤因子-1在肾上皮性肿瘤中的表达及临床意义

目的 明细胞癌、乳头状肾细胞癌、肾嫌色细胞癌、Xp11.2易位/TFE3基因融合相关性肾癌和转移性透明细胞癌中的表达率分别是77.8%(49/63)、90.9%(20/22)、1/13、7/7和87.5%(21/24),7例嗜酸细胞腺瘤均阴性.在原发性肾透明细胞癌中,KIM-1弥漫阳性表达更易发生于Furhman细胞核Ⅲ/Ⅳ级的病例(P=0.010).肾特异性钙黏蛋白主要表达于嫌色细胞癌和嗜酸细胞腺瘤.结论 KIM-1仅表达于损伤的近曲小管和由其起源的肿瘤,对原发性和转移性肾透明细胞癌、乳头状肾细胞癌及Xp11.2易位/TFE3基因融合相关性肾癌具有高度的特异性和敏感性,与肾特异性钙黏蛋白合用可以提高原发性肾脏上皮性肿瘤组织学分类的准确性和转移性肾透明细胞癌的诊断率.     

EZH2和RUNX3在肾细胞癌中的表达及其临床意义

目的 检测EZH2和RUNX3蛋白在肾细胞癌中的表达,探讨它们与肾细胞癌临床病理特征间的关系,并分析它们的表达相关性.方法 利用免疫组织化学法分别检测肾细胞癌标本及对应的癌旁正常肾组织(各56例)中EZH2和RUNX3的表达情况,并利用统计学软件进行统计分析.结果 EZH2和RUNX3蛋白在肾癌组织中的阳性表达率分别为67.9％和37.5％,在癌旁正常肾组织的阳性表达率分别为28.6％和67.3％;EZH2和RUNX3在肾癌组织和癌旁正常组织中的表达差异均有统计学意义(P＜0.05),肾细胞癌中EZH2的高表达和RUNX3的低表达均与临床分期相关(P＜0.05),与病理分化程度、性别、年龄等的关系无统计学差异(P＞0.05).Spearman相关分析显示二者在肾癌中的表达没有明显的负相关关系.结论 EZH2和RUNX3可能共同参与了肾癌的发生发展过程,可成为肾癌的早期诊断及判断预后的指标.     

人锌铁调控蛋白-1(hZIP1)在肾透明细胞癌组织中的表达及意义

目的研究hZIP1蛋白在肾透明细胞癌组织中的表达及其临床意义。方法采用免疫组化及Western blot方法检测86对肾透明细胞癌及癌旁正常肾组织中hZIP1蛋白的表达情况,并结合临床资料分析hZIP1蛋白表达的临床意义。结果 hZIP1蛋白在肾透明细胞癌组织中的表达水平明显低于在癌旁正常肾组织中的表达(P0.01),在Ⅰ+Ⅱ期肾癌组织中的阳性率为80.3%(49/61),显著高于在Ⅲ+Ⅳ期肾癌组织中的阳性率40%(10/25,P0.001),在1+2级肾癌组织中的阳性率为90.3%(56/62),显著高于在3+4级肾癌组织中阳性率12.5%(3/24,P0.001)。结论 hZIP1蛋白可能成为肾透明细胞癌的一个早期诊断指标及治疗的一个重要靶点。     

肾细胞癌的临床病理与免疫表型研究

目的 研究肾细胞癌的临床病理特征、预后及免疫表型特点.方法 复习114例肾细胞癌的临床病理资料、HE切片,按2004年WHO肾肿瘤分类标准重新分类、随访并进行免疫组织化学染色.结果 114例.肾细胞癌包括5个类型,肾透明细胞癌77例(67.5%)、乳头状肾癌11例(9.6%)、肾嫌色细胞癌14例(12.3%)、Xp11.2易位_/TFE3基因融合相关性肾癌10例(8.8%)、未能分类肾肿瘤2例(1.8%).免疫组织化学结果,肾透明细胞癌主要表达CK(93.5%,72/77)、CD10(93.5%,72/77)、波形蛋白(75.3%,58/77),乳头状肾癌主要表达α-甲酰基辅酶A消旋酶(AMACR,11/11),肾嫌色细胞癌主要表达CD117(11/14),Xp11.2易位/TFE3基因融合相关性肾癌TFE3、AMACR、CD10和CK的阳性率分别为10/10、10/10、9/10和7/10.结论 肾癌是一组形态学上各有特征的异质性肿瘤,在形态学基础上,CD10、波形蛋白、CD117、AMACR、CK7、TFE3有助于亚型的诊断.     

青少年肾细胞癌的临床病理及分子遗传学研究

目的 探讨青少年肾细胞癌的临床病理特征、遗传学改变、鉴别诊断及预后.方法 对46例青少年肾细胞癌进行光镜观察及免疫组织化学染色,随访并复习相关文献.对46例肿瘤进行von Hippel-Lindau(VHL)基因区域杂合性缺失(LOH)及VHL基因突变筛查.结果 共诊断19例Xp11.2易位/TFE3基因融合相关性肾癌(Xp11 RCC)、9例透明细胞癌、17例乳头状肾细胞癌(PRCC)和1例不能分类肾细胞癌.19例Xp11 RCC均TFE3阳性,而TFEB阴性.8例肿瘤具有巢状和乳头状结构形态类似t(X;17)ASPL-TFE3型肾癌,6例肿瘤组织学类似t(X;1)PRCC-TFE3型肾癌,4例肿瘤形态像透明细胞癌,1例肿瘤组织学形态文献中未被检索到,表现为细胞核呈毛玻璃样,核仁不明显,可见核沟,肿瘤间质见大量黏液.LOH及VHL突变检测结果显示,仅1例透明细胞癌和1例2型PRCC存在LOH,并且该2型PRCC的VHL基因的一个剪切位点存在胚系突变,553+5 G→C.其余45例均未检测出VHL突变.统计学分析表明TFE3阳性肾细胞癌比TFE3阴性肾细胞癌更倾向于高病理分期(pT3/pT4),并且预后较差(P=0.035).结论 青少年肾细胞癌表现出不同的组织学形态以及分子遗传学背景.其中Xp11 RCC为最常见的肾癌亚型.TFE3阳性肾细胞癌的预后要差于TFE3阴性肾细胞癌.     

肾透明细胞癌中Eotaxin-1的表达与肿瘤增殖能力相关

目的探讨嗜酸性粒细胞趋化因子-1 Eotaxin-1在肾透明细胞癌(ccRCC)中的表达情况及其与肿瘤增殖能力的关系。方法免疫组织化学检测58例肾透明细胞癌患者肾癌组织及癌旁组织中Eotaxin-1以及增殖细胞核抗原(PCNA)的表达并分析其与肾癌临床病理特征的关系;用Western blot法分析10例相互配对肾癌组织、癌旁组织中Eotaxin-1及PCNA的表达,并对Eotaxin-1与PCNA做相关性分析。结果 Eotaxin-1主要表达于胞质,PCNA主要表达于细胞核内,Eotaxin-1和PCNA在肾癌组织的阳性率明显高于癌旁组织(P0.05);病理分级为2-3级组的Eotaxin-1及PCNA阳性率显著高于病理分级1级组(P0.01)。临床分期为Ⅲ/Ⅳ组的Eotaxin-1及PCNA阳性率显著高于临床分期Ⅰ/Ⅱ组(P0.05),有淋巴结转移组的Eotaxin-1及PCNA阳性率高于无淋巴结转移组(P0.05);10例肾癌组织中Eotaxin-1及PCNA蛋白水平显著高于癌旁组织(P0.01,P0.05);Eotaxin-1与PCNA在肾癌组织中的表达存在相关性(P0.05)。结论肾透明细胞癌组织中Eotaxin-1与肾透明细胞癌的增殖能力密切相关,对Eotaxin-1的研究可能为临床对肾透明细胞癌诊断治疗提供新的思路。     

肾透明细胞癌中AnnexinⅡ的表达及其临床意义

目的:探讨AnnexinⅡ在人肾透明细胞癌和癌旁组织样本中的表达情况及其临床意义。方法:采用Western blot方法检测29例新鲜肾透明细胞癌及相应癌旁组织中AnnexinⅡ蛋白的表达;采用免疫组织化学方法检测120例肾透明细胞癌及相应癌旁组织中AnnexinⅡ的表达情况,并将结果与患者的临床病理参数及预后进行相关性分析。结果:AnnexinⅡ在肾透明细胞癌及相应癌旁组织中均有表达,前者的表达水平较后者显著升高(P<0.01);免疫组织化学显示肾透明细胞癌组织中,AnnexinⅡ的阳性表达率高达67.5%,与癌旁组织存在明显差异(P<0.01);AnnexinⅡ在肾透明细胞癌组织中以胞膜表达为主,其表达状况与肾透明细胞癌患者的临床分期(P<0.05),组织学分级(P<0.05),肾被膜受侵(P<0.01)和远地转移(P<0.01)紧密相关;并且AnnexinⅡ的表达状况与肾透明细胞癌患者的5年生存率显著相关。结论:AnnexinⅡ在肾透明细胞癌组织中过表达,其与肾癌的发展密切相关,在肾癌的侵袭及转移过程中可能发挥重要作用。     

ICAM-1和8-OHdG在肾癌组织中的表达及其与临床病理指标的关系

目的:观察ICAM-1和8-OHdG在肾癌组织中的表达情况,并分析两者的表达与肾癌患者临床病理参数之间的关系,探讨其在肾癌发生和进展中的作用。方法:采用免疫组化的方法检测46例肾癌组织和18例远离癌组织的正常肾组织中ICAM-1和8-OHdG的表达,通过阳性染色强度及阳性细胞百分比进行综合评分,观察不同组织者中两者的表达差异,同时分别统计分析ICAM-1和8-OHdG的表达与肾癌患者年龄、性别、肿瘤大小、肿瘤分期以及是否有淋巴结转移或远处转移等临床病理参数之间的关系。结果:ICAM-1在正常肾组织中的表达多为弱阳性或阴性表达,而在肾癌组织中ICAM-1阳性表达增加(占比82. 6%),且与肿瘤的分化程度、淋巴结转移和远处转移具有显著相关性(P0. 05或P0. 01)。绝大多数正常肾组织中8-OHdG呈阴性表达,而肾癌组织中8-OHdG阳性表达增加(占比63%),与肿瘤大小、淋巴结转移和远处转移以及无进展生存期(PFS)之间具有显著相关性(P0. 05或P0. 01)。结论:ICAM-1和8-OHdG在肾癌组织中高表达,二者在肾癌的发生和进展中可能发挥着重要的作用。     

miR-497与肾癌预后的关系及其对肾癌786-0细胞增殖、凋亡和侵袭的影响

目的:探讨微小RNA-497(miR-497)与肾癌预后的关系及其对肾癌786-0细胞增殖、凋亡和侵袭的影响。方法:收集2011年~2015年80例肾细胞癌患者手术切除的癌组织和癌旁组织并回顾性分析患者随访资料。采用实时定量PCR法检测癌组织及配对的癌旁组织中miR-497的表达。在肾癌细胞系786-0中转染miR-497模拟物,应用MTT法、台盼兰拒染法活细胞计数、流式细胞术和Transwell小室实验检测miR-497对786-0细胞增殖、凋亡和侵袭能力的影响。通过生物信息学预测miR-497在786-0细胞中作用的靶基因。Western blotting法检测miR-497对靶基因蛋白表达影响。结果:肾癌组织中miR-497的表达量明显低于癌旁组织(P0.05)。786-0中miR-497表达量明显低于正常肾上皮细胞中miR-497表达量(P0.05)。随访时间2~48个月,中位随访时间29个月,失访6例,随访率92.5%。76例患者3年无复发生存率(recurrence-free survival,RFS)为55.2%。高miR-497表达组和低miR-497表达组肾癌患者的RFS分别为71.2%和40.1%,差异有统计学显著性(P0.05)。过表达miR-497可显著抑制786-0细胞的增殖和侵袭能力,显著促进细胞凋亡(P0.05)。过表达miR-497可显著抑制786-0细胞中cyclin D1蛋白的表达(P0.05)。结论:miR-497与肾癌预后相关,miR-497能明显抑制肾癌786-0细胞的增殖和凋亡,其机制可能与其靶向抑制cyclin D1有关。     

CDKNA2A mutation analysis, protein expression, and deletion mapping of chromosome 9p in conventional clear-cell renal carcinomas: evidence for a second tumor suppressor gene proximal to CDKN2A

Inactivation of tumor suppressor genes on chromosome 9p is considered a critical event in renal cell carcinoma pathogenesis. Alterations of CDKN2A on 9p21 have been reported in renal cancer cell lines, but their relevance for primary renal carcinomas is unclear. Loss of heterozygosity (LOH) was analyzed by using four polymorphic microsatellites at D9S970 (9p12-9p13), D9S171 (9p13), D9S1748 (9p21), and D9S156 (9p21) in 113 primary conventional clear-cell renal cell carcinomas (CRCCs). Allelic deletion was detected in 21 of 88 informative CRCCs (24%) with the highest rate of LOH being observed at D9S171 on 9p13 (20%). Chromosome 9p LOH was associated with short tumor-specific survival in stage pT3 RCC (P = 0.01). Fluorescence in situ hybridization analysis of 54 CRCCs revealed no homozygous CDKN2A deletions indicating that this mechanism of CDKN2A inactivation is rare in CRCC. Sequencing of 113 CRCCs showed that 13 tumors (12%) had a 24-bp deletion abrogating codons 4 through 11 of CDKN2A. Immunohistochemical CDKN2A expression was absent in normal renal tissue and was only detected in six of 382 CRCCs (1.5%) on a renal tumor microarray. These data suggest that CDKN2A alterations are present in a small subset of CRCCs and a second, yet unknown tumor suppressor gene proximal to the CDKN2A locus, may play a role in CRCC development.     

新疆维吾尔族妇女宫颈上皮内瘤变及宫颈癌中FHIT、P16~(INK4a)、RARβ蛋白的表达及意义

目的探讨脆性组氨酸三联体(fragile histidine triad,FHIT)、P16INK4a与视黄酸受体β(retinoic acid receptor-beta,RARβ)蛋白在新疆维吾尔族妇女宫颈上皮内瘤变(cervical intra-epithelial neoplasia,CIN)及宫颈癌中的表达及其意义。方法采用免疫组化链霉素抗生物素蛋白-过氧化物酶链接(SP)法检测20例慢性宫颈INK4a炎、30例CIN(CINⅠ、CINⅡ、CINⅢ各10例)以及40例浸润性宫颈鳞癌组织标本中FHIT、P16INK4a及RARβ蛋白的表达。结果(1)FHIT蛋白阳性表达率依次为慢性宫颈炎(90.00%)CIN(66.67%)浸润性宫颈鳞癌INK4a(27.50%)(P=0.000);P16和RARβ蛋白阳性表达率依次为浸润性宫颈鳞癌(82.50%;90.00%)CIN(46.67%;53.33%)慢性宫颈炎(0.00%;10.00%)(P=0.000;P=0.000)。(2)在宫颈各组织中,FHIT蛋白INK4a与P16INK4a蛋白表达呈负相关(r=-0.384,P=0.000);FHIT蛋白与RARβ蛋白表达呈负相关(r=-0.291,P=0.006);P16INK4a蛋白与RARβ蛋白表达呈正相关(r=0.445,P=0.000)。结论抑癌基因FHIT表达缺失与P16INK4a和RARβ过度表达与宫颈癌的发生发展密切相关,并且FHIT、P16INK4a、RARβ具有协同作用。FHIT蛋白与P16INK4a蛋白和RARβ蛋白的联合检测可作为宫颈癌早期诊断和宫颈癌进展的分子指标。     

P选择素在几种恶性肿瘤中表达的临床意义

研究P选择素在胃癌、肾癌、肺癌、胶质瘤及卵巢肿瘤中表达的临床意义。应用免疫组化LSAB法分别检测P选择素在上述肿瘤中的表达。结果显示P选择素在上述肿瘤中阳性表达率分别为胃癌 5 3 3% (32 / 60例 )、肾癌 5 5 % (11/ 2 0例 )、肺癌 5 4% (2 7/ 5 0例 )、胶质瘤 61 5 % (2 4/ 39例 )、卵巢恶性肿瘤 5 3 3% (16/ 30例 )。P选择素不仅在肿瘤血管内皮上表达 ,亦在肿瘤细胞膜上表达。胃癌患者P选择素阳性者平均生存期和 5年生存率显著低于阴性者。此外 ,III、IV期肿瘤患者P选择素阳性表达率显著高于I、II期患者。P选择素与上述肿瘤的浸润转移相关 ,并可能是一种新的肿瘤转移及预后指标。     

乳癌组织端粒酶活性表达及其与p16基因的关系

目的：探讨乳房恶性肿瘤组织中端粒酶活性的表达情况及ｐ１６抑癌基因的影响。方法：应用端粒重复放大程序－酶标法（ＴＲＡＰ－ＥＬＩＳＡ）及ＴＲＡＰ－银染法检测３５例有乳癌组织及２４例良性肿瘤组织中的端粒酶活性,并采用多重ＰＣＲ技术对乳癌组织的ｐ１６基因的缺失空谈进行了分析,结果：３５例乳癌标本中有２８例端粒酶活性表达为阳性（８０％）,而２４例乳房良性肿瘤标本中仅１例端粒酶活性表达阳性（４．１％）。此外３５例乳癌标本中的１５例存在有ｐ１６基因的外显子２及外显子１的缺失（４２．８％）,而该１５例中有１４例被检测为端粒酶活性阳性（９３．３％）,但２０例未缺失标本的端粒酶阳怀率仅为７０％（１４／２０）。结论：端粒酶活性与乳房肿瘤组织的恶性程度密切相关,提示端粒酶参与了肿瘤的形成过程,而端粒酶活性的增高与ｐ１６抑癌基因的缺失突     

Characterization of a 3;6 translocation associated with renal cell carcinoma

The most frequent cause of familial clear cell renal cell carcinoma (RCC) is von Hippel-Lindau disease and the VHL tumor suppressor gene (TSG) is inactivated in most sporadic clear cell RCC. Although there is relatively little information on the mechanisms of tumorigenesis of clear cell RCC without VHL inactivation, a subset of familial cases harbors a balanced constitutional chromosome 3 translocation. To date nine different chromosome 3 translocations have been associated with familial or multicentric clear cell RCC; and in three cases chromosome 6 was also involved. To identify candidate genes for renal tumorigenesis we characterized a constitutional translocation, t(3;6)(q22;q16.1) associated with multicentric RCC without evidence of VHL target gene dysregulation. Analysis of breakpoint sequences revealed a 1.3-kb deletion on chromosome 6 within the intron of a 2 exon predicted gene (NT_007299.434). However, RT-PCR analysis failed to detect the expression of this gene in lymphoblast, fibroblast, or kidney tumor cell lines. No known genes were disrupted by the translocation breakpoints but several candidate TSGs (e.g., EPHB1, EPHA7, PPP2R3A RNF184, and STAG1) map within close proximity to the breakpoints.     

Expression of retinoblastoma protein and P16 proteins in classic Hodgkin lymphoma: relationship with expression of p53 and presence of Epstein-Barr virus in the regulation of cell growth and death

Deregulation of several genes involved in cell cycle control has been reported in classic Hodgkin lymphoma (cHL). This study aimed to investigate the expression of tumor suppressor proteins (P16(INK4A), retinoblastoma protein, and p53) in cHL in relation to the proliferation and apoptosis of Hodgkin/Reed-Sternberg (H/RS) cells, correlating with the status of Epstein-Barr virus (EBV). A total of 66 cHL cases and 10 nonneoplastic reactive lymphoid tissues were retrieved from the archives. Immunohistochemistry technique was used for the detection of protein expression. Presence of EBV infection was detected by EBV early RNA in situ hybridization. p16(INK4A) gene deletion status was assessed by fluorescence in situ hybridization technique. Expression of P16(INK4A) was observed in 49.2% of the cases, whereas positive retinoblastoma protein and p53 expressions in the H/RS cells were detected in 89.1% and 81.5% of the cases, respectively. Epstein-Barr virus positivity was detected in 53.0% of the cases. Proliferation marker, Ki-67 expression, was observed in 86.7% of the cases. There was no significant correlation between the expression of the various tumor suppressor proteins and Ki-67. Retinoblastoma protein and p53 were also not associated with the presence of EBV. An inverse relationship was observed between the expression of P16(INK4A) and the presence of EBV. There were no significant homozygous or hemizygous deletions of the p16(INK4A) gene. However, an aberrant copy number of chromosome 9 with the loss of one or more p16(INK4A) loci was detected in all cases assessable by fluorescence in situ hybridization. Loss of function of one or more tumor suppressor proteins may be involved in defective cell regulation of H/RS cells. Epstein-Barr virus may have a role in inhibiting P16(INK4A) expression, thus resulting in a perturbed p16(INK4A)-Rb cell cycle checkpoint.     

Proliferating cell nuclear antigen but not p53 or human papillomavirus DNA correlates with advanced clinical stage in renal cell carcinoma

In this study we investigated 56 renal cell carcinomas immunohistochemically for the expression of proliferating cell nuclear antigen (PCNA) and tumour suppressor protein p53 . We also analyzed for the presence of human papilloma virus (HPV) DNA subtypes 6, 11, 16, 18, 31 and 33 by in situ hybridization. In carcinomas which showed more than 10% of PCNA positive nuclei there were significantly more cases with invasion ( P = 0.032) or metastatic disease ( P = 0.047). Nine out of 22 grade III-IV tumours (40.9%) but only six out of 30 grade I-II tumours (20%) showed more than 10% of PCNA positive cells ( P = 0.097). Patients with 10% or more PCNA positive cells in kidney tumours had more advanced disease at the time of diagnosis than those showing less PCNA positive cells ( P = 0.05).
Six p53 positive cases were found among 56 tumours (11%), but only one case had more than 10% positive cell nuclei. The presence of HPV DNA was found in 29 out of 56 cases (52%). Multiple subtypes were found in 19 cases (34%). The most commonly occurring subtypes were 18 and 33. There was no association between PCNA, p53 and the presence of HPV DNA subtypes. Because of the association of PCNA with invasion and metastatic disease, it would be worth while to study PCNA further as a possible marker for aggressiveness of renal carcinomas. Both this study and those concentrated on mutational analysis suggest that p53 is generally not important for the development of renal cell carcinoma. On the other hand, the presence of HPV DNA in these tumours implicates HPV viral infection in the aetiology of renal cancer.