Staufen基因在结直肠癌组织中的表达

为研究甲基化差异相关基因Staufen在结直肠癌不同组织中的表达情况,采用RT-PCR和免疫组织化学等方法在结直肠癌病人中检测Staufen基因在腺癌、癌旁粘膜和相应远端切缘正常组织中的表达。研究发现,在mRNA水平上,远端切缘正常组织中Staufen基因的表达水平显著高于癌旁粘膜和腺癌（P<0.05）;从正常组织、癌旁粘膜到腺癌,Staufen基因的表达有降低的趋势;未发现性别、年龄、肿瘤部位（结肠/直肠）、分化程度、淋巴结转移等临床病理因素与Staufen基因的表达有关（P均>0.05）。正常组织与腺癌组织的Staufen蛋白的表达水平显著高于癌旁粘膜（P<0.05）,而正常组织与腺癌组织之间未发现有统计学差别（P>0.05）。结果表明,Staufen基因在癌旁粘膜和肿瘤中有低表达的趋势,该基因可能在结直肠癌的发生、发展中起作用。     

小肠腺癌组织中Notch1和P53免疫反应性增强

目的研究小肠腺癌中Notch1、Numb、P53和P63蛋白的免疫组织化学表达情况及意义。方法采用免疫组织化学S-P法检测含有60例小肠腺癌(small intestinal adenocarcinoma,SIA)组织芯片中Notch1、Numb、P53和P63蛋白的表达情况,与9例正常小肠组织进行对照,并进行统计学分析。结果在小肠腺癌组织中,Notch1免疫组织化学染色阳性率显著高于正常小肠组织,Notch1蛋白在小肠腺癌中的免疫组织化学表达与肿瘤TNM分期相关,而与年龄和性别不相关。P53在小肠腺癌组织中的免疫组织化学表达为41.7%,P53在正常小肠组织中无表达。Numb蛋白在小肠腺癌组织和正常小肠组织中阳性表达率分别为68.3%和77.8%,二者比较无统计学差异。Numb蛋白在小肠腺癌组织表达缺失率为31.7%。P63在小肠腺癌组织中表达率为5%,而在正常小肠组织中不表达。P53和Numb表达均与年龄、性别、TNM分期等临床病理参数不相关。相关分析显示,小肠腺癌中Numb与Notch1,P53与Numb、Notch1表达之间无相关性。结论小肠腺癌组织中存在Notch1的过表达,且与小肠腺癌的进展相关,部分病例Numb蛋白表达缺失及P53突变,Numb-Notch1信号通路蛋白可能参与了小肠腺癌的发生机制。     

胆管癌组织中KAI1/CD82基因的表达及意义

目的:探讨肿瘤转移抑制基因KAI1/CD82在胆管癌组织中的表达情况及临床病理意义。方法:应用免疫组织化学技术检测48例胆管癌组织及8例正常胆管组织中的KAI1/CD82蛋白表达。结果:KAI1/CD82蛋白在胆管癌组织中阳性表达率31.3%,明显低于正常胆管组织(87.5%,P<0.01)。KAI1/CD82蛋白的表达与肿瘤分化程度、转移相关(P<0.05),而与胆管癌患者年龄、性别、肿瘤部位和病理类型无关。结论:KAI1/CD82蛋白低表达可能参与了胆管癌的发生、发展,并对肿瘤转移的判断有一定指导意义。     

Neuronatin在人体常见肿瘤中的表达及对肿瘤细胞增殖的影响

目的:研究印记基因Neuronatin(NNAT)在人体常见肿瘤中的表达情况及其两种编码产物NNATα和NNATβ对肿瘤细胞增殖的影响。方法:运用组织芯片免疫组化技术对一些人体常见肿瘤及其对应正常组织中NNAT的表达进行系统检测;同时利用腺病毒载体技术,将NNATα和NNATβ分别导入人结肠癌细胞系SW620,并用实时细胞分析仪和平板克隆实验分析细胞增殖能力的变化。结果:①免疫组化结果显示:NNAT在食道鳞癌,结肠腺癌,直肠腺癌,胰腺腺癌,乳腺非特殊性浸润性导管癌,宫颈鳞癌,子宫内膜癌,膀胱移行上皮癌,肺鳞癌,皮肤鳞癌以及前列腺腺癌11种肿瘤组织,肾上腺,皮肤,睾丸,脑,骨骼肌,胰腺6种正常人体组织,以及慢性结肠黏膜炎与慢性肝炎2种炎性组织中呈阳性。②体外细胞增殖实验结果显示,NNATα对人结肠癌肿瘤细胞增殖有一定的抑制作用。结论:NNAT基因在多种人体肿瘤组织中表达,其α片段的表达对于结肠癌细胞系SW620增殖具有轻微抑制作用。     

印记基因PEG10 在葡萄胎组织中的表达以及意义

目的:通过免疫组化方法,探讨印记基因PEG10在葡萄胎组织中的表达及其在早期鉴别葡萄胎妊娠中的应用价值。方法:选取经病理组织学诊断为完全性葡萄胎、部分性葡萄胎、正常早孕、难免流产的标本共计156例,采用免疫组织化学技术检测PEG10在其中的表达,研究遗传印记基因PEG10在葡萄胎妊娠以及非葡萄胎妊娠中的表达。结果:PEG10在四组蜕膜组织中均有表达,在难免流产组呈弱阳性表达,在正常早孕组呈弱阳性和中度阳性表达,在部分性葡萄胎组中呈中度阳性和强阳性表达,在完全性葡萄胎组中呈强阳性表达。PEG10在葡萄胎妊娠组表达明显增多于非葡萄胎妊娠组,两组比较具有显著性差异(P0.01),部分性葡萄胎组表达增多于难免流产组,两组比较差异有显著性(P0.05)。结论:遗传印记基因PEG10在葡萄胎组织中的表达明显高于正常早期妊娠和难免流产组,PEG10基因表达上调与葡萄胎的发生可能有重要关系,是否可将其用于病理诊断鉴别困难时的辅助手段。     

Cyclin G1、MDM2和p53在胃腺癌组织中的表达及相关性研究

目的探讨细胞周期蛋白G1(cyclinG1)、鼠双微体基因(MDM2)和p53在胃腺癌组织中的表达意义及相关性。方法采用免疫组织化学方法(SP法)检测54例胃腺癌组织中cyclinG1、MDM2和p53的表达,以20例正常胃粘膜组织作为对照。结果cyclinG1、MDM2、p53在胃腺癌组织中的阳性表达率分别为62.96%、53.70%、44.44%,而在正常胃粘膜组织中的阳性表达率分别为0%,15.00%,0%,两者比较差异均有显著性(P均<0.05),cyclinG1、MDM2在胃腺癌中的表达与肿瘤的分化程度相关(P均<0.05)。胃癌组织中MDM2蛋白的表达与p53的表达呈正相关(r=0.307),而cyclinG1蛋白的表达与p53的表达无明显相关性。结论CyclinG1、MDM2、p53的阳性表达在胃癌的发生、发展过程中起着重要的的作用。MDM2可能是通过调控p53的活性而促进胃癌的发生、发展,cyclinG1可能以不依赖p53的途径发挥作用。     

Nucleostemin与MDM2在肺腺癌组织中的表达及意义

核干细胞因子(Nucleostemin,NS)是细胞增殖的调节因子,在肺腺癌组织中的表达及与鼠双微染色体2(murine double minute 2,MDM2)表达的相关性不清楚.通过反转录聚合酶链反应(RT-PCR)检测22对肺腺癌和癌旁正常组织NS mRNA的表达;应用免疫组织化学SP法检测45例肺腺癌和22例癌旁正常组织NS与MDM2蛋白的表达,分析其相关性及意义.研究发现,肺腺癌组织中NS mRNA的相对表达强度明显高于癌旁正常组织(P0.01).NS和MDM2蛋白在肺腺癌组织中的阳性表达率分别为73.3%(33/45)、57.8%(26/45),而癌旁正常组织中无阳性表达(P0.01).二者的阳性表达均与组织学分级相关(P0.05),并且表达呈正相关(P0.05),而与患者性别、年龄、肿瘤大小、TNM分期及淋巴结转移无关(P0.05).结果表明NS基因的高表达对肺腺癌细胞的恶性增殖发挥了重要作用,可能是通过p53通路对细胞周期影响所实现的.     

Id-1在喉癌组织中的表达

目的:证实Id-1基因在喉癌组织和喉癌细胞系中的表达.方法:通过反转录PCR(RT-PCR)和蛋白质印迹杂交法(Westernblot法)对30例喉癌组织,10例癌旁正常组织和一个喉癌细胞株在基因和蛋白水平上对ID-1的表达进行检测.结果:喉癌组织和喉癌细胞株中Id-1基因高表达,而正常喉组织中没有Id-1的表达;Id-1蛋白的表达与喉癌组织的、临床分期、分化程度具有相关性(均P<0.05).结论:ID-1蛋白的表达与喉癌的发生、发展及临床分期有关.     

Nucleostemin基因在非小细胞肺癌组织中的表达及意义

目的探讨Nucleostemin(NS,核干细胞因子)基因在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达及临床意义。方法利用RT-PCR法检测13对NSCLC组织和癌旁正常组织中NS mRNA的表达;采用免疫组织化学SP法检测73例NSCLC组织和13例癌旁正常组织中NS蛋白的表达,并分析与NSCLC患者临床病理特征的关系。结果 NSCLC组织中NS mRNA相对表达强度(0.848±0.305)显著高于癌旁正常组织(0.153±0.020)(t=8.712,P0.01)。NS蛋白在NSCLC中的表达率为58.9%(43/73)显著高于癌旁正常组织中的表达率0%(0/13)(χ2=15.315,P0.01)。NS蛋白的表达率与NSCLC的组织类型及分化程度相关,腺癌组织的表达率为76.5%(26/34)明显高于鳞癌组织的表达率43.6%(17/39)(χ2=8.113,P0.01);低分化组织的表达率81.5%(22/27)明显高于高、中分化组织的表达率45.7%(21/46)(χ2=9.023,P0.01),而与患者性别、年龄、肿瘤大小、TNM分期及淋巴结转移无关(P0.05)。结论 NS基因mRNA及蛋白在NSCLC组织中高表达,对肿瘤细胞的恶性增殖起了重要作用,是一个新的有应用价值的肿瘤分子标志物。     

MDM2基因和TBX2在子宫内膜样腺癌中的表达及临床意义

目的:探讨MDM2和TBX2基因在正常增殖期子宫内膜、子宫内膜增殖症和子宫内膜样腺癌组织中的表达和临床意义。方法:采用免疫组织化学链菌素亲生物素基因过氧化物酶连接法(SP法)和组织芯片技术检测20例增殖期子宫内膜、41例子宫内膜增生性病变和45例子宫内膜样腺癌中MDM2和TBX2基因的表达情况。结果:MDM2在增殖期子宫内膜和单纯性增生子宫内膜中均无强阳性表达,复杂性增生子宫内膜和子宫内膜样腺癌中MDM2强阳性表达率分别为23.81%和51.11,明显高于增殖期子宫内膜和单纯性增生子宫内膜(P<0.05)。子宫内膜样腺癌中MDM2的强阳性表达率为51.11%(23/45),明显高于复杂性增生子宫内膜和复杂性非典型增生中的子宫内膜(P<0.05)。子宫内膜样腺癌中MDM2强阳性表达与肿瘤分化程度和TNM分期密切相关,Ⅱ、Ⅲ级子宫内膜样腺癌的强阳性表达率明显高于I级(72.41%vs 13.33%,P<0.05),而与淋巴结转移无相关性(P>0.05)。TBX2在增殖期子宫内膜,单纯性增生子宫内膜,复杂性增生子宫内膜,子宫内膜样腺癌中的阳性表达率分别为30%,35%,45%和72.7%。子宫内膜样腺癌中TBX2基因的阳性表达明显高于其它各组(P<0.05)。TBX2与子宫内膜样腺癌分化程度、TNM分期均有相关性(P<0.05),与有无淋巴结转移均无相关性(P>0.05)。结论:MDM2、TBX2基因在复杂性增生宫内膜和子宫内膜样腺癌中表达明显增强,提示两者在子宫内膜样腺癌发生中起到一定作用。     

MT1F mRNA在结肠癌组织中的表达及其临床病理意义

目的:探讨MT1F mRNA在结肠癌组织中的表达及其临床病理意义。方法:采用Taq Man探针实时荧光定量Real-time PCR检测40例结肠癌及对应正常粘膜组织中MT1F mRNA的表达,并通过免疫组化检测Metallothionein(MT)的蛋白表达。结果:82.5%(33/40例)的结肠癌组织MT1F mRNA表达较对应正常组织明显下调,降低2.4倍至113倍不等。MT1F mRNA水平与结肠癌患者的年龄、性别、肿瘤部位、肉眼形态、直径、分化及Dukes分期无关。在MT1F mRNA低表达的病例中,癌组织MT蛋白表达低于对应正常组织(P0.05)。结论:结肠癌组织MT1F mRNA水平显著下调,但与结肠癌的临床病理特征无关。     

The relationship between metallothionein-1F (MT1F) gene and hepatocellular carcinoma

To investigate the expression of MT1F gene in hepatocellular carcinoma tissue and the growth suppression effect of exogenous introduction of MT1F gene on liver cell line HepG2 and to explore the potential application of MT1F gene in gene therapy of tumor. Eukaryotic expression vector of pCMV-MT1F plasmid was introduced into HepG2 line which expressed no MT1F protein originally with lipofectamine transfection method. The cell growth curve, soft agar colony formation rate and tumorigenicity in SCID mice were examined to demonstrate the growth suppression effect of exogenous MT1F gene on HepG2 cell line. The MT1F mRNA and MT1F protein were also detected in 60 pairs of surgical specimens of hepatocellular carcinoma by in situ hybridization and immunohistochemistry. The transfected HepG2 cell line grew more slowly than control HepG2 as shown by cell growth curves, the soft agar colony formation rate (3.8 percent vs. 7.4 percent, p <.01) and the average growth rate of tumor in SCID mice (30.9 +/- 6.9 vs. 70.3 +/- 5.6, p <.01). The expression level of MT1F mRNA and protein significantly increased in paracancerous tissue, normal tissue than in cancer tissues (75 percent, 70 percent vs. 16.7 percent by ISH and 66.7 percent, 60 percent vs. 10 percent by IHC, p <.01). Exogenous MT1F gene shows the strong effect of growth inhibition on HepG2 cell line. In the liver cancer tissue, MT1F shows down-regulated expression that supports the inhibited function of MT1F in cancer growth and suggests MT1F may have an important role in gene therapy of hepatocellular carcinoma.     

miR-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为研究

目的:探讨mi R-199a-3p负调控CBX7影响肺癌细胞NCI-H460的生物学行为。方法:qRT-PCR法检测并比较肺癌组织、癌旁正常组织、肺癌细胞、正常肺上皮细胞中的mi R-199a-3p m RNA相对表达量。比较远处转移肺癌组织、未转移肺癌组织中mi R-199a-3p m RNA相对表达量。qRT-PCR法、Western Blot法检测并比较肺癌组织、癌旁正常组织中的CBX7 m RNA及蛋白的表达水平。荧光素酶活性法检测mi R-199a-3p与靶基因CBX7的结合。比较mi R-199a-3p模拟物转染组与阴性对照组的肺癌细胞中的CBX7 m RNA相对表达量及CBX7蛋白表达水平。CCK8实验检测mi R-199a-3p对肺癌细胞增殖的促进作用。Tranwell实验检测mi R-199a-3p对肺癌细胞侵袭与迁移能力的影响。结果:肺癌组织中mi R-199a-3p明显高于癌旁正常组织,发生远处转移的肺癌组织中mi R-199a-3p m RNA的表达量明显高于未发生转移的肺癌组织,差异有统计学意义(P<0.001)。肺癌组织中CBX7m RNA、CBX7蛋白表达水平均明显低于癌旁正常组织,差异有统计学意义(P<0.001)。荧光素酶活性法证实mi R-199a-3p可与靶基因CBX7结合抑制CBX7的表达。肺癌细胞中mi R-199a-3p m RNA的相对表达量明显高于正常肺上皮细胞,CBX7 m RNA相对表达量明显低于正常肺上皮细胞(P<0.05)。对于肺癌细胞,mi R-199a-3p模拟物转染组的CBX7 m RNA相对表达量及CBX7蛋白表达水平均明显低于阴性对照组(P<0.001)。CCK8实验证实mi R-199a-3p能够促进肺癌细胞的增殖,Tranwell实验证实mi R-199a-3p对肺癌细胞侵袭与迁移具有积极的促进作用。结论:mi R-199a-3p在肺癌的发生发展过程中发挥重要作用,能够通过抑制CBX7基因的表达,促进肺癌细胞的增殖、侵袭和转移。     

抑癌候选基因NDRG2在甲状腺癌组织中的表达及其意义

目的:研究抑癌候选基因NDRG2在人类甲状腺癌组织及其癌旁组织中的表达情况.方法:收集30例甲状腺癌组织及其癌旁组织,提取总RNA,应用半定量RT-PCR方法检测NDRG2 mRNA的表达水平.分别提取30例组织的总蛋白,应用蛋白印迹技术检测其NDRG2的蛋白表达水平.结果:RT-PCR结果显示,30例甲状腺癌组织中,有25例NDRG2的mRNA水平明显降低,蛋白印迹结果显示,30例甲状腺癌组织中发现25例NDRG2的蛋白水平明显下降,与RT-PCR检测结果一致.结论:NDRG2在甲状腺癌组织中呈低表达,提示其可能对甲状腺癌的发生或发展有重要作用影响.     

Association of NDRG1 Gene Promoter Methylation with Reduced NDRG1 Expression in Gastric Cancer Cells and Tissue Specimens

NDRG1 (N-myc downstream-regulated gene 1) plays a role in cell differentiation and suppression of tumor metastasis. This study aims to determine the expression of NDRG1 mRNA and protein in gastric cancer cell lines and tissue specimens and then assess the possible cause of its aberrant expression. Six gastric cancer cell lines and 20 pairs of normal and gastric cancer tissue samples were used to assess NDRG1 expression using Real-time PCR and Western blot. High-resolution melting analysis (HRM) and methylation-specific PCR (MSP) were performed to detect gene mutation and methylation, respectively, in cell lines and tissues samples. Expression of NDRG1 mRNA and protein was downregulated in gastric cancer cell lines and tissues. Specifically, expression of NDRG1 mRNA and protein was lower in all six gastric cancer cell lines than that of normal gastric cells, while 15 out of 20 cases of gastric cancer tissues had the reduced levels of NDRG1 mRNA and protein. HRM data showed that there was no mutation in NDRG1 gene, but MSP data showed high levels of NDRG1 gene promoter methylation in the CpG islands in both cell lines and tissue samples. Moreover, treatment with the DNA methyltransferase inhibitor 5-Aza-2′-deoxycytidine upregulated NDRG1 expression in gastric cancer HGC27 cells, but not in the histone deacetylase inhibitor trichostatin A-treated HGC27 cells. In conclusion, this study has shown that expression of NDRG1 mRNA and protein was reduced in gastric cancer cell lines and tissues, which is due to methylation of NDRG1 gene promoter. Further study will unearth the clinical significance of the reduced NDRG1 protein in gastric cancer.     

BRCA1 gene expression in breast cancer: a correlative study between real-time RT-PCR and immunohistochemistry.

Breast cancer is a major cause of cancer-related mortality in women. There are major discrepancies concerning the usefulness of various antibodies in detecting breast cancer susceptibility gene 1 (BRCA1) protein and its subcellular localization. The aim of the present study was to determine the specificity and sensitivity of immunohistochemistry (IHC) as a screening method for demonstrating BRCA1 expression. BRCA1 gene expression in archival paraffin-embedded breast cancer tissues was studied simultaneously at the protein and mRNA levels, and the two findings were compared. Forty-eight archival paraffin-embedded breast cancer tissues were studied for BRCA1 gene expression at protein level by IHC using four different antibodies against different BRCA1 epitopes and at mRNA level using real-time RT-PCR. BRCA1 mRNA expression was reduced or absent in 79% of the samples, and this finding correlated significantly with loss of BRCA1 protein expression in 83% of breast cancer tissues using one BRCA1 antibody studied (AB-1, against N-terminus epitope). The specificity of this antibody was 91.3%, and its sensitivity was 66.6%. There was no significant correlation between BRCA1 mRNA and protein expression as demonstrated by the remaining three antibodies. Antibody 8F7 had the highest sensitivity of 100%, but its specificity was 30.4% if mRNA levels were considered as the reference standard.     

前列腺癌患者组织中RPL6的表达及临床意义

目的:探讨前列腺癌患者组织中核糖体蛋白L6(RPL6)表达,分析其对肿瘤恶性程度的评估作用。方法:收集2013年12月-2015年12月在本院接受治疗的前列腺疾病患者117例,根据病理结果分为前列腺炎组63例、前列腺癌组54例;另取同期进行健康体检的健康者80例作为正常对照组。采用Western-blot法检测三组研究对象的前列腺组织RPL6表达,采用酶联免疫吸附法(ELISA)测定血清肿瘤标志物含量,采用RT-PCR法检测前列腺组织增殖基因、侵袭基因mRNA表达,采用Pearson检验分析RPL6表达量与肿瘤恶性程度的相关关系。结果:前列腺癌组患者的前列腺组织RPL6表达量高于前列腺炎组及正常对照组(P0.05);前列腺癌组患者的血清前列腺特异性抗原(PSA)、前列腺特异性酸性磷酶(PSAP)、尿激酶型纤溶酶原激活剂(u PA)、硫氧还蛋白(TRX)含量高于前列腺炎组及正常对照组(P0.05);前列腺癌组患者的前列腺组织增殖基因URG11、PTEN mRNA表达量高于前列腺炎组及正常对照组,PRDM5 mRNA表达量低于前列腺炎组及正常对照组,侵袭基因IgGHG1、TMPRSS2-ER、PIK3C2B mRNA表达量高于前列腺炎组及正常对照组(P0.05)。前列腺癌患者的组织RPL6表达量与肿瘤标志物含量、增殖及侵袭基因活性均呈直接相关关系(P0.05)。结论:高表达的RPL6是前列腺癌早期诊断的可靠指标,且与肿瘤恶性程度直接相关,有望成为前列腺癌辅助诊断及预后评估的重要手段。     

Elevated cyclin E level in human clear cell renal cell carcinoma: possible causes and consequences

The expression of cyclin E gene (CCNE) in relation to the expression of its major regulatory protein, E2F1, was examined in clear cell renal cell carcinomas (ccRCC). We show that the overexpression of E2F1 is accompanied by the significant increase of the mean amounts of cyclin E mRNA, as well as of total cyclin E protein and its low molecular weight forms in cancer tissues as compared to peritumoral controls. A significant increase of the mean amount of total cyclin E was found in peritumoral tissues compared to cancer-free kidneys, suggesting that cancer cells might secrete factors having a profound influence on the metabolism of neighbouring tissues. A significant, positive correlations between E2F1 protein and total cyclin E mRNA, as well as between E2F1 protein and full length cyclin E protein were found in cancer-free kidneys and in peritumoral tissues, but not in ccRCCs. The overexpression of cyclin E positively correlated with the decreasing degree of tumor differentiation, implicating a role for cyclin E in the promotion of tumorigenesis.