Menin蛋白和TGF-β在甲状旁腺肿瘤中的表达及临床意义

目的探讨Menin蛋白与转化生长因子-β(transforming growth factorβ,TGF-β)在甲状旁腺肿瘤发生中的作用及相互关系。方法62例标本包括正常甲状旁腺组织7例,甲状旁腺增生组织12例,甲状旁腺腺瘤组织43例。通过免疫组化法检测它们中Menin蛋白和TGF-β的表达,并进行分组比较。结果Menin蛋白的表达,正常组、增生组同腺瘤组比较差异均有统计学意义(P<0.05),正常组同增生组比较差异无统计学意义(P>0.05)。TGF-β的表达,正常组、增生组和腺瘤组三者之间比较差异均无统计学意义(P>0.05)。但Menin蛋白和TGF-β在腺瘤中的表达存在显著相关(P<0.05)。结论Menin蛋白的失活在甲状旁腺肿瘤的发生中起重要作用;Menin是TGF-β信号传递途径中的一种关键蛋白。     

PTEN蛋白与cyclin-D1蛋白在结肠癌中的表达及意义

目的 :研究人第10号染色体缺失的磷酸酶及张力蛋白同源的蛋白(PTEN蛋白)、细胞周期蛋白D1(cyclin-D1)在结肠癌中的表达及临床意义。方法:用免疫组化SP法检测78例结肠癌患者癌组织及癌旁正常黏膜组织中PTEN蛋白与cyclin-D1的表达情况,分析两者在结肠癌发生、发展过程中的表达情况及相互关系。结果:(1)PTEN蛋白在结肠癌组织中低表达,在正常黏膜组织高表达(P0.05),并随着肿瘤分化程度的降低、临床分期的提高、浸润深度的加深、淋巴结的转移而降低(P0.05)。(2)cyclin-D1在结肠癌组织中高表达,在正常黏膜组织低表达(P0.05),并随着肿瘤分化程度的降低、临床分期的提高、浸润深度的加深、淋巴结的转移而升高(P0.05)。(3)PTEN蛋白及cyclin-D1在肿瘤细胞中的表达呈负相关。结论:PTEN蛋白表达下调、cyclin-D1上调与结肠癌的发生、发展、浸润、转移有关,对两者的检测有助于对结肠癌患者做出病情评估及预测预后。     

早期直肠癌中p53和hMLH1的表达

目的探讨p53基因和hMLH1基因的突变是否是直肠癌发生的早期事件之一。方法用PV-9000二步法免疫组化检测技术对我院2003年2月至2009年7月期间的32例早期直肠癌、32例直肠腺瘤及30例正常直肠黏膜组织石蜡切片进行p53和hMLH1基因蛋白表达的检测。结果①正常直肠黏膜组织、直肠腺瘤组织及早期直肠癌组织中p53蛋白表达阳性率分别为0(0/30)、59.38%(19/32)及68.75%(22/32),其在直肠腺瘤组织和早期直肠癌组织中的p53蛋白表达阳性率差异无统计学意义(P0.05),但二者均明显高于其在正常直肠黏膜组织中的表达(P0.01);hMLH1蛋白在前述三种组织中的表达阴性率分别为0(30/30)、12.50%(4/32)和50.00%(16/32),其在早期直肠癌组织中的表达阴性率明显高于在直肠腺瘤组织和正常直肠黏膜组织中的表达(P0.01)。②在直肠腺瘤组织和早期直肠癌组织中p53阳性表达同时合并hMLH1阴性表达者分别占9.38%(3/32)及37.50%(12/32),二者间差异有统计学意义(P=0.008)。③p53阳性表达和hMLH1阴性表达均与早期直肠癌的分化程度有关(P0.05),而均与患者的性别、年龄、肿瘤最大径、浸润深度及大便潜血均无关(P0.05)。④p53阳性表达与腺瘤的增生程度有关(P=0.009),hMLH1阴性表达与腺瘤的增生程度无关(P0.05)。结论 p53基因和hMLH1基因的同时突变致癌作用明显,可能是直肠癌发生的早期事件之一。     

生长激素受体在直肠癌组织中的表达

目的 研究生长激素受体(GHR)在直肠癌患者癌组织和正常直肠组织中的表达情况.方法 收集接受直肠癌根治术且手术断端无肿瘤转移患者的直肠癌标本43例.按癌细胞分化程度分成高、中、低3组,癌组织及远端断端正常直肠组织用免疫组化法行GHR染色.结果 中分化组1例直肠癌患者断端正常直肠黏膜GHR染色为可疑阳性,其余患者癌组织及断端直肠黏膜GHR染色均为阳性.低分化组直肠癌组织中GHR的表达水平明显高于中分化组和高分化组(P<0.05),中分化组GHR表达亦高于高分化组,但无统计学意义(P>0.05);高分化组断端正常直肠黏膜GHR表达水平显著低于中分化组和低分化组(P<0.05),中分化组正常直肠黏膜GHR表达低于低分化组,但无统计学差异(P>0.05).各组癌组织与断端正常直肠黏膜GHR表达无显著性差异(P>0.05).结论 直肠癌患者癌组织和断端正常直肠粘膜的GHR表达水平基本一致,随着直肠癌细胞分化程度的降低,GHR在癌组织及断端正常直肠黏膜中的表达有增高的趋势.     

神经因子诱导的克隆B基因和Nur相关因子1基因在醛固酮瘤中的表达及意义

目的 探讨神经因子诱导的克隆B(NGFI-B)基因和Nut相关因子1(Nurr1)基因在醛固酮瘤中的表达及意义.方法 应用实时逆转录-聚合酶链反应(Real time RT-PCR)检测34例醛固酮瘤和13例正常肾上腺组织中神经因子诱导的克隆B基因,Nur相关因子1基因,醛固酮合成酶基因(CYP11B2)和皮质醇合成酶基因(CYP11B1)的mRNA表达水平.结果 醛固酮腺瘤组织中NGFI-B基因的mRNA表达量与正常肾上腺组织的差异无统计学意义(P>0.05),醛固酮腺瘤组织中Nurr1基因呈高表达,与正常肾上腺组织的差异有统计学意义(P<0.01),醛固酮腺瘤组织中Nurr1基因的表达量与其CYP11B1和CYP11β2的表达量之间明显相关(分别r=0.434,P<0.01;r=0.376.P<0.05).结论 孤核受体Nurr1对醛固酮瘤组织中CYP11B2的表达有促进作用,醛固酮瘤组织CYP11B1和CYP11B2的表达是多种转录因子共同作用的结果.     

明胶酶A及其抑制剂、碱性成纤维细胞生长因子及受体在肺鳞癌的表达

目的 探讨明胶酶A(MMP2)及其抑制剂组织金属蛋白酶抑制物(TIMPs),碱性成纤维细胞生长因子(bFGF)及其受体(FGFR)-1在人肺鳞癌(SCCL)中的表达与肿瘤转移及患者预后的相关性。方法采用免疫组织化学链霉抗生物素蛋白-过氧化物酶(SP)法检测30例有转移的肺鳞癌、20例无转移的肺鳞癌、15例正常支气管上皮、12例增生上皮和10例不典型增生上皮中MMP2、TIMP2和bFGF、FGFR-1蛋白的表达。结果 正常支气管上皮→增生上皮→不典型增生→无转移鳞癌→有转移鳞癌的演进过程中,MMP2、TIMP2和bFGF、FGF-1蛋白阳性表达率均逐步显著增高(P<0.05)。有转移癌与无转移癌相比,MMP2、TIMP2和bFGF表达的差异均有显著性(P<0.05),增生 不典型增生上皮与正常上皮相比,MMP2表达的差异具有显著性(P<0.05)。4种蛋白的表达率均与鳞癌TNM分期正相关(P<0.05)。Kaplan meier分析发现MMP2和TIMP2阳性表达患者的术后生存率显著低于阴性表达者(P<0.05)。结论 MMP2、TIMP2和bFGF、FGFR-1过度表达导致肿瘤的浸润转移及预后不良。     

组织芯片技术检测人睾丸基因TDRG1在睾丸肿瘤组织中的表达

目的:探讨人睾丸基因TDRG1在睾丸肿瘤组织中的蛋白表达及其病理学意义。方法:运用组织芯片技术、免疫组化法检测人睾丸特异基因TDRG1在睾丸肿瘤组织和正常睾丸组织中的表达。结果:15例正常人睾丸对照组中11例(73.3%)TDRG1蛋白表达为阳性;26例精原细胞瘤中7例(26.9%)TDRG1蛋白表达为阳性,7例畸胎瘤中4例(57.1%)TDRG1蛋白表达为阳性。而12例胚胎癌中10例(83.3%)TDRG1蛋白表达为阳性,10例卵黄囊瘤中8例(80.0%)TDRG1蛋白表达为阳性。精原细胞瘤实验组与正常人睾丸对照组相比较,两组间具有极显著性差异(P<0.01)。畸胎瘤实验组与正常人睾丸对照组相比较,两组间具有显著性差异(P<0.05)。而胚胎癌组和卵黄囊瘤组与正常人睾丸对照组相比较,没有显著性差异(P>0.05)。结论:TDRG1蛋白在精原细胞瘤和畸胎瘤的表达水平较正常对照睾丸组织显著降低,TDRG1可能为候选的抑癌基因。     

TGF-α与EGFR在继发性甲状旁腺功能亢进症腺体组织中的表达及意义

目的:探讨转化生长因子α(TGF-α)与表皮生长因子受体(EGFR)在继发性甲状旁腺功能亢进症(SHPT)患者甲状旁腺腺体组织中的表达及意义。方法:收集36例SHPT组织标本以及7例正常甲状旁腺组织标本,HE染色区分弥漫性增生和结节性增生甲状旁腺,用免疫组化法检测所有甲状旁腺组织中TGF-α、EGFR和Ki-67的表达,并分析TGF-α、EGFR表达与SHPT患者临床病理因素的关系。结果:36例SHPT中弥漫性增生17例,结节性增生19例,甲状旁腺的增生类型与患者的年龄、性别、透析时间、术前甲状旁腺激素水平、血钙、血磷、钙磷乘积、血清白蛋白、血红蛋白、尿素氮、肌酐和碱性磷酸酶水平均无明显关系(均P0.05),TGF-α、EGFR、Ki-67的表达在正常甲状旁腺织、弥漫性增生甲状旁腺织、结节性增生甲状旁腺织中均依次明显升高(均P0.05)。结论:TGF-α与EGFR在SHPT患者腺体组织中表达增高,且与甲状旁腺细胞的增生方式密切有关。     

大肠癌及腺瘤中端粒酶催化亚单位的检测及其临床意义

目的探讨端粒酶催化亚单位(hTERT)在大肠癌发生中的作用及其在大肠癌早期诊断中的意义。方法应用逆转录聚合酶链反应(RTPCR)的方法对10例正常肠黏膜、40例腺瘤及21例早期大肠癌中的hTERTmRNA进行定量检测。结果肿瘤和中、重度不典型增生腺瘤中hTERTmRNA表达量明显高于轻度不典型增生腺瘤(P<0.01);hTERTmRNA在肿瘤组织和中小腺瘤(≤2cm)中的表达量差异有显著性(P<0.05)。结论利用RTPCR的方法检测大肠肿瘤组织中hTERTmRNA的表达敏感、可行,hTERT有望成为一个有效的大肠癌早期诊断的参考指标。     

β-链接素与基质金属蛋白酶-7在大肠腺瘤及其癌变组织表达的意义

目的探讨β链接素(β-catenin,β-cat)和基质金属蛋白酶-7(matrixmetalloproteinase7,MMP-7)表达与大肠腺瘤癌变的关系。方法应用组织芯片和免疫组化技术检测β-cat和MMP-7在25例大肠正常黏膜及135例腺瘤及腺瘤癌变组织的表达。结果(1)腺瘤癌变组织β-cat胞质、胞核表达率分别为68.4%、40.4%,显著高于腺瘤(40.0%、16.0%)及正常粘膜(4.0%、0.0%,均为P<0.01)。(2)高级别与低级别上皮内肿瘤相比β-cat胞质、胞核表达率显著升高(均为P<0.05),而胞膜表达率无显著性差异(P>0.05)。(3)正常黏膜、腺瘤、腺瘤癌变组织MMP-7阳性表达率依次增高,腺瘤癌变组织MMP-7阳性表达率(45.6%)显著高于正常黏膜(16.0%,P<0.05)。(4)大肠腺瘤及其癌变组织中β-cat胞质、胞核表达与MMP-7阳性表达均呈正相关关系(均为P<0.001)。结论β-cat胞质/胞核表达及MMP-7阳性表达与大肠腺瘤癌变的发生有关;β-cat与MMP-7的协同表达可能在腺瘤恶性转化过程中发挥着重要的作用。     

Expression of PRAD1/cyclin D1, retinoblastoma gene products, and Ki67 in parathyroid hyperplasia caused by chronic renal failure versus primary adenoma

BACKGROUND: In primary hyperparathyroidism, certain genetic abnormalities responsible for parathyroid tumorigenesis are proposed, and it has been reported that the overexpression of PRAD1/cyclin D1 induced by a DNA rearrangement of the parathyroid hormone (PTH) gene is one of the genetic disorders in a number of primary parathyroid adenomas. However, in secondary hyperparathyroidism caused by uremia, the mechanism of monoclonal proliferation in nodular parathyroid hyperplasia is not well understood. To elucidate the mechanism, we examined the expression of PRAD1/cyclin D1, retinoblastoma gene products, and Ki67 in primary adenoma and secondary hyperplasia. METHODS: In adenomas (N = 15) and associated glands (N = 7) with normal histology obtained from patients with primary hyperparathyroidism and in diffuse (N = 14), multinodular (N = 58), and single nodular (N = 28) glands from patients who underwent parathyroidectomy for renal hyperparathyroidism, the expression of these cell cycle regulators was evaluated by immunohistochemical technique. A labeling index was used to define the proportion of cells with positive nuclear staining by each antibody. RESULTS: In 6 out of 15 (40%) primary adenomas, PRAD1/cyclin D1 was overexpressed (a labeling index of more than 500), possibly because of the PTH gene rearrangement, but not in secondary hyperplasia, including single nodular glands. Compared with diffuse hyperplasia, nodular hyperplasia showed a significantly higher expression of PRAD1/cyclin D1 (P < 0.05), retinoblastoma gene products (P < 0.05), and Ki67 (P < 0.05). However, no statistically significant correlation between the expression of PRAD1/cyclin D1 and that of Ki67 was observed in both primary adenoma and secondary hyperplasia. CONCLUSIONS: These results suggest that in secondary hyperplasia caused by uremia, at least remarkable overexpression of PRAD1/cyclin D1 induced by PTH gene rearrangement may be not the major genetic abnormality responsible for tumorigenesis. Heterogenous genetic changes seem to contribute to monoclonal proliferation of parathyroid cells induced by the expression of PRAD1/cyclin D1 or by some other mechanism independent of the amplification of the proto-oncogene.     

应用生物信息学方法结合细胞实验探究 Plakophilin-1（PKP1）在皮肤黑色素瘤中的生物学作用

目的 探索PKP1在皮肤黑色素瘤（skin cutaneous melanoma，SKCM）中相关生物学功能。 方法 通过实时TIME 2.0数据库分析PKP1在不同的肿瘤以及正常组织中的相对表达量；通过GEPIA以及 UALCAN数据库分析PKP1在皮肤黑色素瘤组织以及正常组织的表达量以及探究PKP1的表达水平和肿瘤 病人预后的关系；在SK-MEL-1细胞系中，过表达PKP1探究对于肿瘤细胞的影响。结果 TIME 2.0分析显 示，多种肿瘤组织中PKP1表达水平低于正常组织（P＜0.05）；而胆管癌、肝癌以及肾透明细胞癌等肿瘤 中PKP1表达水平高于正常组织（P＜0.05）；GEPIA数据库分析显示，正常组织中PKP1表达水平高于皮肤 黑色素瘤组织（P＜0.05）；PKP1过表达组PKP1的蛋白水平高于NC组（P＜0.05）；PKP1高表达组OS指标 优于低表达组（P＜0.05）；CCK8实验显示，PKP1过表达组与空质粒组细胞OD值比较，统计学意义显著 （P＜0.01）。结论 PKP1在皮肤黑色素瘤中低表达，其表达水平下调和预后不良有关，PKP1可以抑制皮肤 黑色素瘤细胞的增殖，PKP1有成为靶向治疗的关键基因的潜力。     

EphrinA1在原发性肝细胞癌中的表达及临床意义

目的探讨原发性肝细胞癌中EphrinA1的mRNA和蛋白表达水平及其临床意义。方法应用逆转录聚合酶链反应（RT-PCR）及免疫组织化学方法检测EphrinA1的mRNA及蛋白在40例肝癌及相应的癌旁肝组织和10例正常肝组织中的表达水平,并分析其与临床病理学特点之间的关系。结果40例肝癌组织及相应的癌旁肝组织和10例正常肝组织中均有EphrinA1 mRNA的表达。RT-PCR分析显示EphrinA1mRNA在肝癌组织（0．5413±0．1527）中的表达显著高于癌旁肝组织（0．3895±0．0549,P〈0．05）和正常肝组织（0．3770±0．1055,P〈0．05）,而在癌旁肝组织（0．3895±0．0549）和正常肝组织（0．3770±0．1055）中的表达差异无统计学意义（P〉0．05）。从正常肝组织、癌旁肝组织到肝癌组织,EphrinA1蛋白的阳性表达率分别为20％（2／10）、35％（14／40）和62％（25／40）,呈递增趋势（x^2=14．762,P〈0．05）。EphrinA1蛋白的过表达与肝癌细胞的分化程度、门静脉癌栓的形成及淋巴结转移等临床病理因素有关（P〈0．05）。结论EphrinA1蛋白的过表达与肝癌细胞的分化程度、淋巴结转移和门静脉癌栓有密切联系,提示其可能在肝癌的恶性转化、侵袭和转移过程中发挥重要作用。     

FOXN2在前列腺癌组织的表达及对前列腺癌细胞生物学的影响

探讨双头框蛋白N2（FOXN2）在前列腺癌（PCa）组织中的表达以及对PCa细胞生物学的影响。方法 选取宜宾市第二人民医院50例接受手术治疗的PCa患者的标本及癌旁组织,通过RT-qPCR实验和Western blot实验分别检测PCa组织和癌旁组织中的FOXN2 mRNA及蛋白的表达水平,并分析PCa组织中FOXN2与患者临床病理特征的相关性。通过RT-qPCR实验检测正常前列腺细胞RWPE-1、PCa细胞PC-3、DU145、LNCaP中FOXN2 mRNA的表达水平。选取PC-3细胞为研究对象,分为FOXN2过表达组、空载体对照组以及对照组,分别通过MTT实验、流式细胞实验、Transwell实验以及Western blot实验检测各组细胞增殖、凋亡、迁移和侵袭情况以及相关蛋白Bax、CyclinD1、MMP-2的表达量。结果 与癌旁组织相比,FOXN2的mRNA和蛋白表达水平显著降低（P<0.05）,与TCGA数据库中结果一致。FOXN2低表达与PCa患者淋巴转移、TNM分期以及Gleason评分有关（P=0.003、0.005、0.002）。与人正常前列腺细胞RWPE-1相比,FOXN2在PCa细胞PC-3、DU145、LNCaP中均呈现低表达（P<0.05）,其中PC-3细胞中表达量最低。与对照组和空载体对照组相比,FOXN2过表达组的PC-3细胞在作用24 、48 、72 h后增殖能力显著下降（F=290.400、57.735、113.014,P<0.05）,CyclinD1蛋白表达水平显著下降（P<0.05）;PC-3细胞的凋亡率显著升高（P<0.05）,Bax蛋白表达水平显著升高（P<0.05）;PC-3细胞的迁移和侵袭能力显著下降（P<0.05）,MMP-2蛋白表达水平显著下降（P<0.05）。结论 FOXN2在PCa组织及细胞中低表达,过表达FOXN2可抑制PCa细胞的增殖、迁移和侵袭,并促进细胞凋亡。     

多聚唾液酸神经细胞黏附分子在垂体腺瘤中的表达及其临床意义

目的 探讨垂体腺瘤中多聚唾液酸神经细胞黏附分子(PSA-NCAM)的表达及与侵袭性、组织学类型、Ki-67抗原之间的关系.方法 收集61例手术切除的垂体腺瘤标本及临床资料,3例正常垂体腺组织来自24 h内死亡的尸解标本,应用免疫组织化学法检测PSA-NCAM在不同类型腺瘤中的表达.结果 本组61例垂体瘤中有26例(52.6%)PSA-NcAM阳性表达,3例正常垂体组织皆为阴性.25例男性患者中11例(44.0%)PSA-NCAM阳性表达,36例女性患者中15例(41.7%)PSA-NCAM阳性表达,两者差异无统计学意义(P>0.05).PSA-NCAM阳性表达组的平均年龄(42.1±11.2)岁,阴性表达组为(44.7±12.4)岁,两者差异无统计学意义(P>0.05).PSA-NCAM阳性表达组的平均Ki-67抗原标记指数(Ki-67 LI)为(3.6±1.3)%,阴性表达组为(2.7±1.4)%,两者差异有统计学意义(P<0.05).PSA-NCAM阳性表达组的肿瘤平均宽度为(2.5±0.9)cm,阴性表达组的为(1.6±1.2)cm,差异有统计学意义(P<0.05).侵袭性垂体腺瘤的PSA-NCAM阳性表达率62.5%(20/32),非侵袭组20.7%(6/29),两者差异有统计学意义(P<0.01).结论 PSA-NCAM的表达与垂体腺瘤的大小、侵袭性、增殖指数密切相关,而与患者的性别、年龄等无明显相关.     

Apoptosis in parathyroid hyperplasia of patients with primary or secondary uremic hyperparathyroidism

BACKGROUND: Chronic oversecretion of parathyroid hormone (PTH) is associated with parathyroid hyperplasia, reflecting a disturbed balance between cell proliferation and apoptosis. This study addressed the unsolved issue of apoptosis in hyperparathyroidism. METHODS: Parathyroid glands from 19 patients with primary (1 degrees ) and 11 patients with secondary (2 degrees ) uremic hyperparathyroidism, as well as 13 normal parathyroid glands, were examined. Apoptosis was evaluated by terminal deoxynucleotidyl transferase (Tdt)-mediated dUTP nick end-labeling assay (TUNEL). Because the apoptotic process is regulated by several oncoproteins, the expression of Bcl-2 and Bax was analyzed by immunohistochemistry. RESULTS: The numbers of apoptotic cells in 1 degrees parathyroid adenoma (0.99 +/- 0.03 per 1000 cells, mean +/- SE, P < 0.009) and 2 degrees parathyroid hyperplasia (1.20 +/- 0.54 per 1000 cells, P < 0.005) were significantly higher than in normal parathyroid tissue (0.13 +/- 0. 06 per 1000 cells). Light microscopy examination of hyperplastic parathyroid tissue from a uremic patient showed the presence of nuclei with dense chromatin characteristic of apoptosis. Bcl-2 staining was strong in normal tissues but weak or negative in several sections of 1 degrees and 2 degrees hyperparathyroid tissues, mostly in nodular areas. Bax staining was homogeneous in normal tissue but patchy in several hyperplastic tissues. CONCLUSION: These results suggest that hyperparathyroidism is associated with a compensatory increase in apoptosis, possibly favored by a diminished Bcl-2/Bax ratio. This renders highly improbable the hypothesis that parathyroid hyperplasia is due to a decreased rate of apoptosis.     

Prognostic significance of molecular alterations in human pancreatic carcinoma – an immunohistological study

Background: During the last decade, many molecular alterations have been described for pancreatic carcinomas. However, the clinical and prognostic value of these alterations has been discussed and is controversial. Methods: An immunhistochemical study was performed in 82 cases of adenocarcinoma of the pancreas. Using specific antibodies, expression of EGF, EGF-receptor, cERB-B2, p53, p21^CIP1, cyclin-D1, BCL-2, CD95 and KI67 was evaluated. Results: Overexpression of the different molecules was found in 44–69% of the pancreatic carcinomas. With regard to clinico-pathological features, p53 positivity was more frequently found in advanced and undifferentiated tumours (P<0.05), EGF overexpression was significantly more frequent in advanced tumours (P<0.05) and CD95 overexpression was observed to a greater extent in undifferentiated tumours (P<0.05). Besides cyclin-D1, none of the molecules tested was of prognostic significance. Patients whose tumours expressed cyclin-D1 lived significantly shorter than patients with cyclin-D1-negative tumours. However, in subgroup analyses of patients with the same tumour stage or tumour grade, even cyclin-D1 expression had no prognostic significance. Conclusion: These results demonstrate that the prognostic significance of the molecules tested here is low. Nevertheless, with regard to tumorigenesis and tumour biology of pancreatic carcinoma, determination of molecular alterations could provide important information about pancreatic cancer. Received: 16 February 1998     

促肝细胞再生磷酸酶-3基因在结直肠癌组织中的表达及其临床意义

目的检测促肝细胞再生磷酸酶-3(PRL-3)mRNA在结直肠癌(CRC)中的表达,探讨其与CRC发生发展以及侵袭转移的关系。方法半定量PCR测定46例CRC组织及其对应正常黏膜、6例结直肠腺瘤(CRA)及其对应正常黏膜以及18例淋巴结和肝转移灶中PRL-3mRNA相对表达水平,并分析其表达水平与临床病理学指标的关系。单链多态性分析法(PCR-SSCP)检测基因突变情况。结果46例CRC中PRL-3基因表达量较相应正常黏膜组织高,差异有统计学意义(1.6±0.7比0.4±0.1,P<0.01);6例结直肠腺瘤组织与对应的正常黏膜中PRL-3mRNA表达量差异无统计学意义(0.6±0.3比0.5±0.2,P>0.05);12例淋巴结转移灶和6例肝转移灶PRL-3基因表达量分别为2.1±0.8和3.3±1.0,显著高于原发癌肿、正常黏膜、阴性淋巴结组织中的表达(P<0.01)。PRL-3基因表达水平与CRCDukes分期、浸润深度、淋巴结和远处转移有关(P<0.05),与性别、肿瘤大小、分化程度无关(P>0.05)。PCR-SSCP分析发现,6例肝转移灶中有1例出现异常泳动条带。结论PRL-3基因在CRC的发生发展和侵袭转移中起重要作用,其作用可能是通过基因表达上调和基因突变共同实现的。     

大肠癌中Caspase-8和Caspase-10的表达及其与细胞凋亡的关系

目的 探讨大肠肿瘤中Caspase-8和Caspase-10的表达及其与细胞凋亡的关系.方法 采用实时荧光定量逆转录-聚合酶链反应(RT-PCR)检测121例大肠癌、52例腺瘤和121例癌旁组织中Caspase-8和Caspase-10 mRNA表达,原位末端标记法(TUNEL)检测细胞凋亡.结果 与癌旁组织比较,腺瘤中Caspase-8 mRNA表达上调,Caspase-10 mRNA表达下调(P<0.05);大肠癌中Caspase-10 mRNA表达下调(P<0.01),下调程度与肿瘤的分化程度有关(P<0.05).大肠癌、腺瘤和癌旁组织中的凋亡指数分别为1.63%、2.42%和2.98%,各组间差异有统计学意义(P<0.05).大肠癌中凋亡指数与Caspase-10 mRNA表达呈正相关(r=0.408,P<0.01),与Caspase-8mRNA表达无明显相关.结论 Caspase-8 mRNA在腺瘤中表达上调,与肿瘤细胞凋亡无明显相关.Caspase-10与大肠癌的凋亡相关,并与肿瘤恶性程度关系密切.     

Effect of endothelin receptor antagonist on parathyroid gland growth, PTH values and cell proliferation in azotemic rats.

BACKGROUND: A variety of stimuli are involved in the pathogenesis of parathyroid gland hyperplasia in renal failure. Recently, it was shown that blocking the signal from the endothelin-1 (ET-1) receptor (ET(A)R/ET(B)R) by a non-selective receptor antagonist, bosentan, reduced parathyroid cell proliferation, parathyroid gland hyperplasia and parathyroid hormone (PTH) levels in normal rats on a calcium deficient diet. Our goal was to determine whether in 5/6 nephrectomized (NPX) rats with developing or established hyperparathyroidism, the endothelin receptor blocker, bosentan, reduced the increase in parathyroid cell proliferation, parathyroid gland hyperplasia and PTH values. METHODS: High (HPD, 1.2%) or normal phosphorus diets (PD) (NPD, 0.6%) were given to 5/6 NPX rats for 15 days (NPX(15)). In each dietary group, one-half the rats were given bosentan (B) i.p. 100 mg/kg/day. The four groups of rats were: (1) NPX(15)-1.2% P; (2) NPX(15)-1.2% P+B; (3) NPX(15)-0.6% P; and (4) NPX(15)-0.6% P+B. In a second study in which hyperparathyroidism was already established in 5/6 NPX rats fed a HPD for 15 days, rats were divided into two groups in which one group was maintained on a HPD and the other group was changed to very low PD (VLPD, <0.05%) for an additional 15 days. In each dietary group, one-half the rats were given bosentan i.p. 100 mg/kg-day. The four groups of rats were: (1) NPX(30)-1.2% P; (2) NPX(30)-1.2% P+B; (3) NPX(30)-0.05% P and (4) NPX(30)-0.05% P+B. Parathyroid cell proliferation was measured by proliferating cell nuclear antigen (PCNA) staining and ET-1 expression by immunohistochemical techniques. RESULTS: In the study of developing hyperparathyroidism, bosentan reduced ET-1 expression in the parathyroid glands of rats on the NPD and HPD (P<0.05). But only in rats on the NPD did bosentan result in a reduced increase in parathyroid gland weight (P<0.05). In the study of established hyperparathyroidism, in which 5/6 NPX rats were given a HPD for 15 days, bosentan started on day 15 reduced (P<0.05) ET-1 expression in rats maintained for 15 additional days on the HPD or the VLPD. On the VLPD, parathyroid gland weight was less (P<0.05) than that in rats on the HPD sacrificed at 15 or 30 days. Bosentan did not reduce parathyroid cell proliferation or parathyroid gland weight in rats maintained on the HPD or further reduce these parameters beyond that obtained with dietary phosphorus restriction. PTH values were lowest in the VLPD group, intermediate in the NPD group, and highest in the HPD group, but in none of the three groups did bosentan decrease PTH values. CONCLUSIONS: In azotemic rats with developing hyperparathyroidism, bosentan resulted in a reduced increase in parathyroid gland weight when dietary phosphorus content was normal. Despite a reduction in ET-1 expression in rats on a HPD with developing or established hyperparathyroidism, bosentan did not reduce the increase in parathyroid cell proliferation, parathyroid gland growth or PTH values. Thus, ET-1 blockade with bosentan did not prevent parathyroid gland growth in the azotemic rat.