结直肠黏膜光镜下炎症表现及 NF-κB、COX-2的表达与结直肠肿瘤发病的相关性分析

目的：观察结直肠黏膜光镜下炎症表现,探讨细胞核因子（ NF-κB）、环氧合酶（ COX-2）的表达与结直肠肿瘤发病的相关性。方法选取2013年7月至2014年7月收治的30例结直肠癌患者、30例结直肠腺瘤患者、30例健康体检者,取肿瘤组织及距离其5 cm处正常黏膜,对照组取正常黏膜进行检查,利用显微镜观察组织淋巴细胞浸润程度、腺体形态等,并对NF-κB、COX-2表达水平进行检测。结果结直肠癌、癌旁黏膜、直肠腺瘤、腺瘤旁黏膜淋巴细胞浸润程度较正常黏膜更高（ P ＜0．05）;结直肠癌旁黏膜腺体变异率高于腺瘤旁黏膜、正常结肠黏膜（ P ＜0．05）;结直肠癌、结直肠腺瘤患者NF-κB、COX-2表达水平高于正常黏膜（ P ＜0．05）。结论结直肠癌、腺瘤患者,结直肠黏膜存在炎症表现,主要表现为腺体变异、淋巴细胞浸润等,在结直肠癌、腺瘤组织中,NF-κB、COX-2表达水平较高,有利于为疾病的诊断、治疗提供专业参考。     

CUL4相关因子7、微RNA-589-5p在结直肠癌组织中表达与临床病理特征及预后的关系

目的 分析结直肠癌组织中CUL4相关因子7(DCAF7)、微RNA-589-5p(miR-589-5p)表达水平,并探讨其与结直肠癌临床病理特征及预后的关系。方法 纳入2018年9月至2019年9月于宝鸡市人民医院行结直肠癌根治术的78例。采用实时荧光定量PCR法检测组织中miR-589-5p、DCAF7 mRNA表达水平。免疫组化法检测组织中DCAF7表达。Spearman法分析组织miR-589-5p与DCAF7的相关性。分析miR-589-5p、DCAF7水平与结直肠癌临床病理特征的关系。Kaplan-Meier法分析结直肠癌组织miR-589-5p、DCAF7表达与病人预后的关系,多因素Cox回归分析影响结直肠癌预后的因素。结果 Ualcan数据库中,结肠腺癌组织中miR-589表达水平低于癌旁组织（P<0.05）,DCAF7表达水平高于癌旁组织（P<0.05）。与癌旁组织相比,结直肠癌组织中DCAF7阳性表达率及DCAF7 mRNA相对表达水平（2.79±0.84比1.02±0.31）较高（P<0.05）,miR-589-5p相对表达水平（0.44±0.09...     

CPNE7在结直肠癌组织中的表达及临床意义

目的：研究结直肠癌患者肿瘤组织中CPNE7的表达及与患者病情、预后的关系。方法：回顾性选择90例结直肠患者为研究对象,经手术获取结直肠癌组织标本和癌旁正常黏膜组织标本,采用Western blotting免疫印迹法检测标本组织和肿瘤细胞系中CPNE7的表达,采用免疫组化检测结直肠癌组织芯片中CPNE7的表达,分析CPNE7表达水平与肿瘤病理特征的关系及在预后评估中的应用。结果：CPNE7在结直肠癌细胞株中的表达明显高于正常结直肠上皮细胞株,且与癌旁正常组织比较,CPNE7在结直肠癌组织中的表达明显升高。90例患者中共84例(93.33%)出现肿瘤组织中CPNE7表达量高于癌旁组织(P<0.05)。肿瘤组织中CPNE7高表达率为50.00%,高于癌旁组织的7.78%(P<0.05)。淋巴结转移N1、N2者CPNE7高表达率均高于N0者(P<0.05),TNM分期Ⅲ期者CPNE7高表达率高于Ⅰ期、Ⅱ期者(P<0.05)。Kaplan-Meier生存曲线分析发现,CPNE7表达水平与患者预后相关(P<0.05)。结论：结直肠癌患者肿瘤组织中CPNE7的表达较癌旁...     

NF-κB在结直肠癌中的表达及与凋亡的关系

目的探讨核因子-κB(nuclear factor-κB,NF-κB)在结直肠癌组织中的表达及与凋亡的关系。方法采用免疫组织化学SP法、TdT介导的dUTP缺口末端标记技术原位观察65例结直肠癌组织及其对照癌旁正常黏膜组织中NF-κB的表达情况和细胞凋亡情况。结果结直肠癌中NF-κB的阳性率为58.46%(38/65),显著高于对照组(P<0.05)。癌旁和癌组织凋亡指数(AI)分别为(8.23±1.57)%和(3.25±1.82)%,两者比较差异有统计学意义(P<0.05)。NF-κB的表达与癌组织的分化程度、淋巴结转移和Dukes分期相关(P<0.05)。结论 NF-κB在结直肠癌组织中高表达,NF-κB表达可抑制结直肠癌的细胞凋亡,并与病理分期、有无淋巴结转移等恶性临床病理特征有密切相关性,提示NF-κB在结直肠癌的发生、发展及预后中发挥重要作用。     

中心体相关蛋白55、睾丸表达基因14在结直肠癌组织中表达及其临床意义

目的 回顾性分析中心体相关蛋白55(CEP55)、睾丸表达基因14(TEX14)在结直肠癌组织中表达及其临床意义。方法选取2015年10月至2016年10月在南阳市中心医院行手术治疗的78例结直肠癌病人癌组织及其癌旁组织为研究对象。利用Ualcan数据库分析CEP55和TEX14在结直肠癌组织及癌旁组织中的表达情况；实时荧光定量PCR(qRT-PCR)法检测CEP55和TEX14 mRNA表达；免疫组化染色法检测CEP55和TEX14蛋白表达；分析CEP55和TEX14蛋白表达与病人临床病理特征的关系；对病人随访5年，使用Kaplan-Meier法绘制生存曲线，分析CEP55和TEX14蛋白表达与结直肠癌病人5年生存率的关系；使用Cox回归模型分析影响结直肠癌病人预后的因素。结果 Ualcan数据库分析结果显示，结直肠癌组织中CEP55和TEX14表达量均明显高于癌旁组织（P<0.05）。结直肠癌组织中CEP55、TEX14 mRNA水平（2.34±0.70比1.00±0.26、1.57±0.44比0.98±0.25）及蛋白阳性表达率均明显高于癌旁组织（P<0.05）。CE...     

ICAM-1、CD82的表达差异对结直肠癌肝转移影响的临床研究

目的对细胞间粘附分子-1(ICAM-1)、CD82的表达差异对结直肠癌肝转移的影响进行相关研究。方法行外科手术治疗结直肠癌患者68例,以自身癌旁正常黏膜组织为对照组,以自身癌组织黏膜组织为研究组,对比观察CD82和ICAM-1蛋白在癌组织的表达情况;对比分析两组CD82及ICAM-1蛋白的表达差异对结直肠癌肝转移的影响及两组的相互关系。结果 68例结直肠癌患者的癌组织中ICAM-1表达明显高于癌旁组织(P<0.05),但在正常大肠黏膜组织中无表达;CD82在结直肠癌组织中表达则明显低于癌旁组织(P<0.05)。同时,ICAM-1、CD82的表达与结直肠癌临床Dukes分期及转移复发有密切关系(P<0.05)。结论可将ICAM-1的表达作为肿瘤发生、浸润、转移的重要判断因子,ICAM-1和CD82联合检测对有效判定肿瘤的恶性程度以及转移情况等均具有临床指导作用。     

结直肠癌组织中Egr-1的表达及意义

目的:研究Egr-1(early growth responsive gene-1,Egr-1)在结直肠癌组织、癌旁粘膜以及癌旁正常粘膜中的表达差异,分析Egr-1核蛋白表达与结肠癌临床病理特征的关系。方法:应用免疫组织化学方法检测结直肠癌组织、癌旁粘膜以及癌旁正常粘膜中Egr-1的表达。结果:Egr-1表达在结直肠癌组织中显著低于癌旁组织及癌旁正常组织(P0.05);结直肠癌组织中Egr-1表达水平与远处转移密切相关(P0.01)。结论:Egr-1基因在结直肠癌中表达下调,并在结直肠癌的发生和转移中起着一定的作用。     

Nup88和PINCH在结直肠癌中表达的关系

目的:探讨核孔蛋白复合体蛋白(Nup88)和PINCH mRNA在结直肠癌组织和肿瘤细胞株中的表达水平及两者的类系。方法:利用RT-PCR的方法研究Nup88和PINCH mRNA在47例结直肠癌组织、44例癌旁无瘤黏膜、45例近端、44例远端黏膜及6个结肠癌细胞株中的表达水平,用Spearman法对Nup88和PINCH进行相关性分析。结果:Nup88和PINCH mRNA在结直肠癌组织中的表达水平高于癌旁无瘤黏膜、近端及远端黏膜,且两者在上述4组标本中表达水平均呈正相关(rs分别为0.31、0.35、0.46及0.48)。结论:Nup88和PINCH可能在结直肠癌发生发展过程中共同发挥作用。     

结直肠癌组织中MMP-7和SLC12A5表达的检测意义

目的探讨结直肠癌组织中基质金属蛋白酶7(MMP-7)以及溶质载体家族12A5基因表达水平的临床检测意义。方法选取2014年6月至2018年6月在我院病理科采取胃肠肿瘤外科手术切除的60例结直肠癌组织,使用免疫组化法PV-9000两步法检测MMP-7和SLC12A5的表达水平。结果 MMP-7在结直肠癌组织中阳性表达率为81.67%(49/60),明显高于结直肠癌旁正常组织的28.33%(17/60)(χ2=12.341,P<0.05);SLC12A5在结直肠癌组织中阳性表达率为83.33%(50/60),明显高于结直肠癌旁正常组织的31.67%(19/60)(χ2=11.379,P<0.05)。结论结直肠癌组织中具有较高的MMP-7和SLC12A5表达水平,应采取免疫组化法对其表达进行检测。     

Quantitative distribution of mRNAs encoding the 19 human UDP-glucuronosyltransferase enzymes in 26 adult and 3 fetal tissues

The purpose of this study was to generate, by real-time PCR, a quantitative expression level profile of the 19 human UDP-glucuronosyltransferases (UGTs) of subfamilies 1A, 2A and 2B, in 26 adult and 3 fetal tissues, for better understanding of their roles in xenobiotic and endobiotic metabolism. Adult liver contained the highest level of combined UGTs mRNA, and UGT2B4 was the most abundant isoform in this tissue (40% of total). Other well expressed hepatic UGTs, in decreasing order of mRNA level, were 1A9, 2B7, 1A4, 2B10, 1A1, 1A6, 2B11, 2B15, 1A3, 2A3, 2B17 and 2B28. UGT2B4 was by far the most abundant isoform in the fetal liver (90% of total). The combined UGT mRNA expression in both adult and fetal olfactory epithelium was high, about 20% the adult hepatic level. Interestingly, a large developmental change was found in this tissue from high UGT2A1 and UGT2A2 expression in the fetus to UGT1A6 in the adult. The most abundantly expressed UGTs in the small intestine were 2A3, 1A10, 1A1, 1A6 and 2B7, while 1A10 and 2A3 predominated in the colon. The results provide the most comprehensive data to date on the tissue distribution of the human UGTs.     

Notch通路上调miR-223抑制FBXW7表达对结肠癌 HCT116细胞生物学的影响

摘要：目的 探讨microRNAs（miRNAs）对结肠癌HCT116细胞增殖和凋亡的影响及其作用机制。方法 经实时 定量 PCR 和 Western blot 检测 20 例结肠癌组织及癌旁正常组织标本中 FBXW7 mRNA 和其蛋白水平。通过 TargetScan 工具预测、定量 PCR 和荧光素酶活性实验鉴定与 FBXW7 结合的 miRNAs,瞬时转染 miR-223 和其对照 miCtr、抑制剂（Inhibitor）入HCT116细胞后检测FBXW7 mRNA和蛋白水平,CCK-8和流式细胞术分别检测细胞活性 和凋亡率。同时通过 siRNA 下调 Notch3 表达后,检测 miR-223 水平以及细胞凋亡率。结果 结肠肿瘤组织中 FBXW7 mRNA与蛋白含量低于癌旁组织（P＜0.05）,预测并证实miR-223与miR-25能够与FBXW7基因特异结合。 HCT116细胞中瞬时转染miR-223后,FBXW7 mRNA水平和蛋白表达量下调（P＜0.01）,细胞活性增加（P＜0.05）,而 细胞凋亡率降低（P＜0.01）。下调Notch3通路后,miR-223表达水平下降（P＜0.001）,FBXW7 mRNA水平上升（P＜ 0.01）,细胞凋亡率下降（P＜0.05）。结论 Notch通路上调miR-223水平,进而抑制FBXW7基因的表达,最终促进结 肠癌细胞HCT116的增殖并抑制其凋亡。     

Single nucleotide polymorphism discovery and functional assessment of variation in the UDP-glucuronosyltransferase 2B7 gene

OBJECTIVE: UDP-glucuronosyltransferase 2B7 (UGT2B7) plays a central role in the liver-mediated biotransformation of endogenous and exogenous compounds. The genetic basis of interindividual variability in UGT2B7 function is unknown. This study aimed to discover novel gene variants of functional significance. METHODS: Caucasian human livers (n=54) were used. UGT2B7 was resequenced in 12 samples [(six highest and six lowest for the formation of morphine-3-glucuronide (M3G)]. Haplotype-tagging single nucleotide polymorphisms were genotyped in the entire sample set. Samples were phenotyped for mRNA expression. RESULTS: 10 haplotype-tagging single nucleotide polymorphisms were identified and their haplotypes were inferred. Haplotype 4 (-45597G; -6682_-6683A; 372A; IVS1+9_IVS1+10A; IVS1+829T; IVS1+985G; IVS1+999C; IVS1+1250G; 801T; IVS4+185C) (frequency of 0.12) was associated with an increase in enzyme activity and gene expression. The 1/4 and 4/6 diplotypes had higher M3G formation compared with 1/1 (P<0.05) and 2/3 (P<0.01) diplotypes. Diplotypes containing haplotype 4 resulted in a significant 45% average increase in the formation of M3G compared with diplotypes without haplotype 4 (P=0.002). There was also an association between haplotype 4 and increased mRNA expression. IVS1+985A>G, 735A>G, and 1062C>T are the putative functional variants of haplotype 4. We also identified two mRNA splicing variants (UGT2B7_v2 and UGT2B7_v3) splicing out exon 1, 4, 5, and 6 but sharing exons 2 and 3 with the involvement of additional 5' exons. UGT2B7_v2 was detected in all livers tested, but UGT2B7_v3 was present at much lower levels compared with UGT2B7_v2. The UGT2B7 reference sequence mRNA is now named UGT2B7_v1. CONCLUSION: UGT2B7 haplotype 4 is functional and its effects on the biotransformation of UGT2B7 substrates should be tested in controlled clinical trials. Biochemical studies should investigate the functional role of the newly discovered mRNA splicing variants.     

Influence of UGT genetic polymorphism on the interindividual variability in mitiglinide pharmacokinetic in Chinese

The aim of the study is to identify the effect of UDP-glucuronosyltransferase (UGT) genetic polymorphism on the interindividual variability in mitiglinide pharmacokinetics (PK) in Chinese. PK and genetic data were obtained from 42 healthy Chinese volunteers (Han ethnic group) treated with single-dose and multiple-dose oral mitiglinide using liquid chromatography/mass spectrometry and DNA sequencing technologies. And the result showed there were great interindividual variabilities in main PK parameters (AUC, Cmax and CL/F) for multiple oral mitiglinide. The enzyme activity of the carriers of UGT1A3 *2/*4 or UGT2B7-1 T/T was stronger than that of the carriers of other genotypes. These results demonstrate that UGT1A3 and UGT2B7-1 genetic polymorphism may have a significant impact on interindividual variability in the PK of mitiglinide in Chinese.     

Monospecific antipeptide antibodies against human hepatic UDP-glucuronosyltransferase 1A subfamily (UGT1A) isoforms

Expression of UDP-glucuronosyltransferases (UGT) in mammals is thought to be regulated in both a tissue- and developmental-specific manner. Furthermore, induction of genes encoding UGT occurs after exposure to xenobiotics including drugs, environmental pollutants and dietary compounds. In human, isoforms of UGT 1A subfamily catalyze the glucuronidation of a greater proportion of drugs, suggesting that the expression of UGT1A isoforms is responsible for the clearance of a diverse range of drugs. To analyze the expression of human UGT1A isoforms, we have developed polyclonal antibodies against specific peptide regions within the isoforms (UGT1A1, 1A3, 1A4, 1A6 and 1A9). The prepared antipeptide antibodies were found to be highly monospecific for each UGT1A isoform and no cross-reactivity with UGT2B isoforms was detected. Analysis of UGT1A protein levels in hepatic microsomes using these antibodies demonstrated interindividual differential expression of each isoform. These highly specific antipeptide antibodies provide an important tool to analyze tissue distribution and interindividual expression levels of human UGT1As.