Epigenetic changes of pituitary tumor-derived transforming gene 1 in pancreatic cancer

BACKGROUND: Pancreatic cancer is a devastating disease with abnormal genetic changes. The pituitary tumor-derived transforming gene (PTTG) is considered to be implicated in the tumorigenesis of cancers when the gene is epigenetically transformed. In this study, we investigated the relationships between aberrant expression and epigenetic changes of the PTTG1 gene in pancreatic cancer. METHODS: We chose 4 cell lines (PANC-1, Colo357, T3M-4 and PancTu I) and pancreatic ductal adenocarcinoma (PDAC) tissues. After using restriction isoschizomer endonucleases (Msp I /Hpa II) to digest the DNA sequence (5'-CCGG-3'), we performed PCR reaction to amplify the product. And RT-PCR was applied to determine the gene expression. RESULTS: The mRNA expression of the PTTG1 gene was higher in pancreatic tumor than in normal tissue. The gene was also expressed in the 4 PDAC cell lines. The methylation states of the upstream regions of the PTTG1 gene were almost identical in normal, tumor pancreatic tissues and the 4 PDAC cell lines. Some (5'-CCGG-3') areas in the upstream region of PTTG1 were methylated, while some others were unmethylated. CONCLUSIONS: The oncogene PTTG1 was overexpressed in pancreatic tumor tissues and verified by RT-PCR detection. The methylation status of DNA in promoter areas was involved in the gene expression with the help of other factors in pancreatic cancer.

Effect of insulin-like growth factor-1 on pituitary tumor transforming gene in glioma C6 cells

Objective: To investigate the effect of insulin-like growth factor-1 (IGF-1) on pituitary tumor transforming gene (PTTG) in glioma C6 cells. Methods: Glioma C6 cells were divided into four groups: A group, treated without IGF-1; B group, treated with 0.1 ng/mL dose of IGF-1; C group, treated with 1 ng/mL dose of IGF-1; D group, treated with 10 ng/mL dose of IGF-1. PTTG mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR), western blotting was used to detect the expression of PTTG protein. Results: The expressions of PTTG mRNA were 1.370 ± 0.212, 2.198 ± 0.354, 3.452 ± 0.332, and 4.576 ± 0.387 respectively in the four groups, and there was a significantly difference between any two groups (P < 0.01). The protein expressions of PTTG in the four groups were 1.407 ± 0.334, 1.813 ± 0.465, 2.412 ± 0.576, and 3.128 ± 0.665 respectively, and there was a significantly difference between any two groups (P < 0.01). Conclusion: IGF-1 can up-regulate the expression of PTTG significantly in dosage-dependent manner.

IN SITU PCR AND IMMUNOHISTOCHEMICAL STUDIES ON p16 GENE IN PITUITARY ADENOMAS

Objective: To examine the occurrence of p16 gene deletion and to analyze p16 expression on paraffin-embedded human pituitary adenoma specimens. Efforts were made to optimize the technical conditions forin situ PCR. Methods:In situ PCR techniques and inimuno-histochemistry were used. Results: Immunohistochemically, p16-positive tumor cells were observed in all cases with various proportions. The majority of the stromal cells and part of tumor cells was devoid of p16 immunostaining, but signal ofin situ PCR for p16 gene, exon 2, was displayed in these cells. Conclusion: The results implied that p16 gene might not be deleted in these pituitary adenomas. It also indicated thatin situ PCR, both direct and indirect methods, proved feasible and informative to the aim of DNA detection. It is critical to overcome non-specific amplification in directin situ PCR by means of higher annealing temperature, fewer cycle, lower magnesium concentration and stringent washing. A target DNA-deleted sample as the negative control is extremely necessary. For the indirect method, the way to improve the sensitivity is to loosen the conditions for amplification and washing, so that more amplification products are subject to hybridization, and signal detection is facilitated.

Correlation of pituitary tumor transforming gene 1 with proliferation and invasion genes in prostate cancer

Objective:To study the change of pituitary tumor transforming gene 1 (PTTG1) expression in prostate cancer and its correlation with proliferation and invasion genes.Methods: Patients with prostate cancer who underwent radical operation in our hospital between March 2015 and January 2018 were selected as the malignant group of the research, and the prostate cancer lesions were collected;patients who underwent transurethral resection of the prostate due to benign prostatic hyperplasia in our hospital during the same period were selected as the benign group of the research, and the benign prostate lesions were collected. The mRNA expression levels of PTTG1, proliferation genes and invasion genes in the lesions were determined. Results:PTTG1, Survivin, Bcl-2, CyclinD1, GPRC6A, ZEB1, CatB, CatD and PAR-1 mRNA expression in prostate cancer lesions of malignant group were significantly higher than those of benign group whereas CDKN2, p21 and TFPI2 mRNA expression were significantly lower than those of benign group;Survivin, Bcl-2, CyclinD1, GPRC6A, ZEB1, CatB, CatD and PAR-1 mRNA expression in prostate cancer lesions with high PTTG1 were significantly higher than those in prostate cancer lesions with low PTTG1 whereas CDKN2, p21 and TFPI2 mRNA expression were significantly lower than those in prostate cancer lesions with low PTTG1.Conclusion:The PTTG1 gene is highly expressed in prostate cancer lesions and it is closely related to the changes of proliferation and invasion gene expression.

Genomic and transcriptomic analyses reveal selection of genes for puberty in Bama Xiang pigs

The Bama Xiang pig （BMX） Chinese indigenous breed is a famous early-maturing with a two-end black coat To uncover the genetic basis of the BMX phenotype, we conducted comparative genomic analyses between BMX and East Asian wild boars and Laiwu pigs, respectively. Genes under positive selection were enriched in pathways associated with gonadal hormone and melanin synthesis, consistent with the phenotypic changes observed during development in BMX pigs. We also performed differentially expressed gene analysis based on RNA-seq data from pituitary tissues of BMX and Large White pigs. The CTTNBP2NL, FRS2, KANK4, and KATNAL1 genes were under selection and exhibited expressional changes in the pituitary tissue, which may affect BMX pig puberty. Our study demonstrated the positive selection of early maturity in the development of BMX pigs and advances our knowledge on the role of regulatory elements in puberty evolution in pigs.

Cloning,tissue distribution and effects of fasting on pituitary adenylate cyclase-activating polypeptide in largemouth bass

Pituitary adenylate cyclase activating polypeptide （PACAP） has a wide range of biological functions. We cloned the full-length cDNAs encoding PACAP and PACAP-related peptide （PRP） from the brain of largemouth bass （Micropterus salmoides） and used real-time quantitative PCR to detect PRP- PACAP mRNA expression. The PRP-PACAP cDNA has two variants expressed via alternative splicing： a long form, which encodes both PRP and PACAP, and a short form, which encodes only PACAR Sequence analysis results are consistent with a higher conservation of PACAP than PRP peptide sequences. The expression of PACAP-Iong and PACAP-short transcripts was highest in the forebrain, followed by the medulla, midbrain, pituitary, stomach, cerebellum, intestine, and kidney; however, these transcripts were either absent or were weakly expressed in the muscle, spleen, gill, heart, fatty tissue, and liver. The level of PACAP-short transcript expression was significantly higher than expression of the long transcript in the forebrain, cerebella, pituitary and intestine, but lower than that of the long transcript in the stomach. PA CAP- long and PACAP-short transcripts were first detected at the blastula stage of embryogenesis, and the level of expression increased markedly between the muscular contraction stage and 3 d post hatch （dph）. The expression of PACAP-long and PACAP-short transcripts decreased significantly in the brain following 4 d fasting compared with the control diet group. The down-regulation effect was enhanced as fasting continued. Conversely, expression levels increased significantly after 3 d of re-feeding. Our results suggest that PRP- PA CAP acts as an important factor in appetite regulation in largemouth bass.

A Qualitative Model of the Interaction of Sexual Behavior and Hormone Gene Transcription in Male Blue Gourami during Reproduction

In the present study, a model is suggested to describe hormone control in male blue gourami (Trichogaster trichopterus) along the gonadotropic brain-pituitary- gonad axis (BPG axis) and the hypothalamic-pituitary-somatotropic axis (HPS axis). This model is based on the cloning and transcription of genes encoding hormones of the two axes involved in spermatogenesis during blue gourami reproduction. Gene transcription is affected by environmental, biological, and behavioral factors. Mature males were examined in two different stages—nonreproductive in high-density habitats and reproductive in low-density habitats. Based on gene transcription, gonadotropin-releasing hormone 1 (GnRH1) was involved in controlling spermatogenesis (spermatogonia to spermatids) via the BPG axis in nonreproductive and reproductive stages by controlling follicle-stimulating hormone (FSH), 11-ketotestosterone (11KT) and 17β-estradiol (E2). However, GnRH3 had a larger effect during the reproductive stage via the BPG axis (spermatids to sperm) on luteinizing hormone (LH), 11KT, and 17α-hydroxyprogesterone (17P). At the same time, the HPS axis was involved in spermatogenesis via pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide PRP (formerly known as GHRH-like peptide) in the brain, and growth hormone (GH) in the pituitary affected synthesis of insulin-like growth factor 1 (IGF1) in the liver.

Visfatin(NAMPT)affects global gene expression in porcine anterior pituitary cells during the mid-luteal phase of the oestrous cycle

Background The pituitary belongs to the most important endocrine glands involved in regulating reproductive functions.The proper functioning of this gland ensures the undisturbed course of the oestrous cycle and affects the female's reproductive potential.It is believed that visfatin,a hormone belonging to the adipokine family,may regulate reproductive functions in response to the female's metabolic state.Herein we ve rified the hypothesis that suggests a modulatory effect of visfatin on the anterior pituitary transcriptome during the mid-luteal phase of the oestrous cycle.Results RNA-seq analysis of the porcine anterior pituitary cells revealed changes in the expression of 202 genes(95 up-regulated and 107 down-regulated in the presence of visfatin,when compared to the non-treated controls),assigned to 318 gene ontology terms.We revealed changes in the frequency of alternative splicing events(235 cases),as well as long noncoding RNA expression(79 cases)in the presence of the adipokine.The identified genes were associated,among others,with reproductive system development,epithelial cell proliferation,positive regulation of cell development,gland morphogenesis and cell chemotaxis.Conclusions The obtained results indicate a modulatory influence of visfatin on the regulation of the porcine transcriptome and,in consequence,pituitary physiology during the mid-luteal phase of the oestrous cycle.

一例CYP3A5代谢酶变异与垂体前叶功能减退病例报告并文献复习

目的 探讨人细胞色素P450家族成员3A5(cytochrome P450 family member 3A5, CYP3A5)编码基因变异对糖皮质激素的代谢影响以及与垂体前叶功能减退的关系。方法 回顾性选取2021年3月青岛大学附属医院肾病科的一例病理类型为膜性肾病合并IgA肾病的肾病综合征患者的临床资料,并对该病进行文献复习。结果 该患者给予常规剂量他克莫司治疗后,检测其他克莫司血药浓度异常增高,发现患者他克莫司代谢酶CYP3A5编码基因存在纯合突变,导致他克莫司蓄积。同时发现该患者垂体前叶功能减退,考虑患者垂体前叶功能减退与糖皮质激素蓄积并反馈抑制垂体前叶功能有关。经过调整糖皮质激素及他克莫司用量,监测患者甲功三项,指标均逐渐好转。结论 CYP3A5基因多态性引起的CYP3A5代谢酶慢代谢型变异,会导致其代谢药物的蓄积,尤其是糖皮质激素,进而引起包括垂体前叶功能减退在内的一系列临床不良结局。

miR-362-3p调控垂体肿瘤转化基因1抑制口腔鳞状细胞癌侵袭及增殖的研究

目的探讨垂体肿瘤转化基因1(PTTG1)在miR-362-3p作用下对口腔鳞状细胞癌(OSCC)细胞Cal-27、HN-30侵袭以及增殖能力的影响。方法生物信息学在线数据库查询PTTG1在头颈部鳞状细胞癌(HNSCC)中的表达。蛋白质印迹法(Western blot)实验检测PTTG1在Cal-27、HN-30以及HOK细胞系中的表达。划痕愈合实验、Transwell侵袭实验及5-乙炔基-2’脱氧尿嘧啶核苷(EdU)细胞增殖实验检测PTTG1对Cal-27、HN-30细胞迁移、侵袭、增殖的影响。生物信息学在线数据库预测PTTG1的上游miRNA,双荧光素酶实验检测结合情况,实时荧光定量聚合酶链反应(qRT-PCR)检测该miRNA在组织中的表达。结果ENCORI数据库结果显示PTTG1在OSCC组织中表达上调;Western blot实验显示Cal-27、HN-30细胞中PTTG1表达量较HOK细胞中高。转染Si-PTTG1质粒的Cal-27、HN-30细胞的迁移能力、侵袭能力和细胞增殖能力均较对照组降低(P<0.05)。通过网站预测出PTTG1的上游miRNA为miR-362-3p,双荧光素酶实验检测出PTTG1与miR-362-3p存在结合位点;qRT-PCR检测结果显示miR-362-3p在OSCC肿瘤组织中相对于正常组织表达下调(P<0.05);并且敲低miR-362-3p的表达后能够促进敲低PTTG1后的Cal-27、HN-30侵袭和增殖。结论miR-362-3p可通过靶向PTTG1抑制Cal-27、HN-30细胞侵袭、增殖。

猪下丘脑和垂体中生长激素受体、胰岛素样生长因子1型受体的发育性变化

GH和IGF 1可作用于垂体或 /和下丘脑负反馈性地调节垂体GH的分泌 ,而这种负反馈作用必须通过下丘脑或垂体的GHR和IGF 1R来实现。为研究猪垂体GH分泌负反馈调节的发育性变化和品种特点 ,分别在 0、3、2 0、30、90、12 0和 180日龄随机选取纯种雄性二花脸猪和大白猪各 4头 ,屠宰并取下丘脑及垂体 ,用相对定量RT PCR分析下丘脑和垂体GHR和IGF 1RmRNA水平。结果表明 :下丘脑GHRmRNA表达呈明显的时序性变化 ,在 0到 12 0日龄期间呈逐渐上升趋势 ,180日龄时显著下降 (P <0 0 5 ) ,提示在快速生长期 ,GH负反馈调控机制逐渐加强。下丘脑GHRmRNA表达还表现明显的品种间差异 ,在 0到 180日龄期间大白猪均显著高于二花脸猪 (P <0 0 5 ) ;而垂体GHRmRNA表达相对稳定 ,品种和年龄间差异不显著 ,提示GH的负反馈作用位点可能主要在下丘脑。IGF 1R与GHR的表达发育模式不同。下丘脑IGF 1RmRNA的表达相对稳定 ,无显著的年龄、品种间差异 ;而在垂体 ,大白猪和二花脸猪IGF 1RmRNA水平在出生时均较高 ,随后显著下降 (P <0 0 5 ) ,2 0日龄后逐渐上升至 90日龄的较高水平 ,随后再次下降。大白猪垂体IGF 1mRNA表达在 30日龄和 90日龄时显著高于二花脸猪 (P <0 0 5 ) ,而 180日龄时二花脸猪垂体IGF 1RmRNA水平却显著高于大白?

右归饮对肾阳虚大鼠下丘脑-垂体-肾上腺轴钙调素mRNA表达的影响

目的 :从分子水平探讨肾阳虚证的本质以及补肾中药的作用机理。方法 :用醋酸可的松造成肾阳虚模型 ,以逆转录多聚酶链反应 (RT PCR)半定量法 ,测定肾阳虚大鼠下丘脑 垂体 肾上腺轴CaMmRNA的表达 ,以及补肾中药右归饮对其的影响。结果 :肾阳虚大鼠下丘脑、肾上腺CaMmRNA水平升高 ,垂体组织CaMmRNA水平没有显著变化 ,右归饮能够降低下丘脑组织CaMmRNA水平 ,但对肾上腺影响不显著。结论 :下丘脑中CaMmRNA水平升高与肾阳虚证有关 ,下丘脑CaMmRNA水平的降低是补肾中药右归饮改善肾阳虚症状的作用机理之一。

糖皮质激素对下丘脑-垂体-肾上腺轴钙调素基因表达的影响

目的 探讨糖皮质激素醋酸可的松对大鼠下丘脑 垂体 肾上腺轴 (hypothalamus pituitay adrenal,HPAA)钙调素基因表达的影响。方法 以逆转录多聚酶链反应 (RT PCR)半定量法测定大鼠注射醋酸可的松后钙调素 (calmodulin ,CaM)mRNA的表达水平。结果 醋酸可的松诱导大鼠下丘脑、肾上腺CaMmRNA水平升高 ,对垂体组织CaMmRNA水平没有显著影响。

慢病毒介导shRNA抑制垂体腺瘤PTTG基因表达及对细胞增殖和侵袭能力的影响

【目的】观察慢病毒介导的sh RNA对GH3和At T20型垂体腺瘤中垂体肿瘤转化基因(PTTG)表达的抑制作用,以及对肿瘤细胞增殖和侵袭能力的影响及其机制探讨。【方法】筛选最佳抑制效果的干扰序列构建PTTG基因sh RNA的慢病毒表达载体,转染GH3和At T20型垂体腺瘤细胞。流式细胞仪分析细胞转染效率,RT-PCR和Western blot检测细胞PTTG基因m RNA和蛋白的表达水平,MTT和Ed U法分别检测靶向PTTG的sh RNA对肿瘤细胞增殖生长的抑制作用,流式细胞仪检测细胞周期变化。Transwell小室检测细胞侵袭能力的变化,基因芯片筛查干扰后的基因表达谱差异并进行生物信息学分析。【结果】细胞转染表达PTTG基因sh RNA的慢病毒载体后,能显著抑制肿瘤细胞PTTGm RNA和蛋白的表达,与对照组相比有统计学差异(P<0.05),细胞的增殖能力受到明显抑制,且增殖抑制效应具有时间依赖性;细胞生长显著变慢,G0/G1期细胞比例显著增高,克隆形成能力下降,细胞侵袭能力也显著减弱,均与对照组相比有统计学差异(P<0.05)。PI3K等信号通路的基因表达也发生显著变化。【结论】慢病毒介导sh RNA沉默PTTG基因表达可能是通过对PI3K信号通路的调控,抑制了GH3和At T20垂体腺瘤细胞的生长,抑制肿瘤细胞的体外增殖和侵袭能力。

PCNA,PTTG在垂体瘤卒中的表达及与侵袭性的关系

目的:通过对垂体腺瘤卒中的垂体瘤转化基因(PTTG)、增殖细胞核抗原(PCNA)蛋白表达的检测,探讨PTTG和PCNA表达与垂体腺瘤卒中细胞生物学活性的关系。方法:将50例垂体腺瘤手术标本分为卒中组与非卒中组,采用免疫组化SABC法检测PTTG和PCNA蛋白的表达,分析卒中组与非卒中组PTTG和PCNA表达水平的差异性。结果:PTTG蛋白表达在卒中组与非卒中组之间差异无统计学意义(P>0.05);卒中组PCNA蛋白表达较非卒中组显著增高(P<0.05);卒中组侵袭性发生率显著高于非卒中组(P<0.05)。结论:PCNA与PTTG表达失衡可能是垂体腺瘤卒中发生的原因之一,PCNA为垂体腺瘤卒中侵袭性行为的主要调控因子。

不同证候H22荷瘤小鼠垂体一致上调或下调的基因

目的:观察常见不同证候荷瘤小鼠垂体基因表达的特征。方法:采用H22荷瘤小鼠及其标准化四诊及辨证方法,以及GeneChip Mouse Exon 1.0 ST Array等技术,检测荷瘤小鼠早期邪毒壅盛证和气虚证、中期阳气虚证、中晚期气阴阳虚证等4个证候及正常小鼠垂体基因的表达,筛选出不同证候一致上调或下调的基因。结果:荷瘤小鼠一致上调的基因有Alas2、Ube2l6、Sgk、BC055107、Cdh26、Bhlhb2、Nr1d2、Slc39a14、Serpina3n、Ms4a6c、Lcp1、Dhx8、Il1r1、S100a8、S100a9、Cxcl1、Hp、Csf1r、RIKEN cDNA2610307008gene、Phactr3、Mknk1、Aloxe3、Aqp11等23个;一致下调的基因有2310047A01Rik、Hoxa10、Cryaa、Hspa1a、Dub2a、Mpz、Klk1、Naaladl1、Prph1等9个;其中部分基因在一些证候中的表达有显著的改变。结论:肿瘤发生后,H22荷瘤小鼠垂体存在大量基因表达的差异,其中部分与证候有关。

绵羊发情周期不同组织Cry1 mRNA转录水平相对定量研究

研究表明,时钟基因Cry1在哺乳动物季节性繁殖内分泌过程中可能发挥了重要作用。文章以多浪羊(非季节性繁殖)和中国美利奴羊(季节性繁殖)为研究对象,采用实时荧光定量PCR技术对下丘脑-垂体-性腺轴主要组织Cry1在发情周期不同阶段的表达变化情况进行了跟踪检测。结果表明:绵羊Cry1在所检测组织中均有表达,其中松果体和甲状腺Cry1的表达水平较高;不同绵羊品种Cry1在发情周期不同阶段的组织表达谱基本一致,除下丘脑外,卵巢、子宫、松果体、垂体和甲状腺中Cry1的表达水平均是在发情前期达到峰值;卵巢、子宫、垂体和松果体Cry1在发情前期和发情期的表达变化幅度存在显著的品种间差异。研究结果表明:Cry1可能具有启动发情和季节性繁殖的重要作用。

PTTG与c-myc在食管癌中的表达及其相关性

目的探讨食管鳞癌组织中垂体肿瘤转化基因(PTTG)和原癌基因(c-myc)的表达和相关性。方法采用免疫组织化学SP法检测PTTG和c-myc在食管癌组织和癌旁正常食管组织中的表达情况。结果48例食管癌组织中PTTG和c-myc的阳性表达率分别为72.9%(35/48)和67.7%(32/48),显著高于癌旁正常食管组织中的表达(分别为10.4%和14.6%,P<0.05),PTTG和c-myc的表达与有无淋巴结转移、TNM分期显著相关(P<0.05),与患者年龄、性别、瘤体大小和肿瘤组织分化程度无相关性;PTTG和c-myc的表达呈正相关(P<0.05)。结论PTTG和c-myc蛋白在食管癌中具有高表达,两者可能参与了食管癌的发生,在食管癌的侵袭和转移中发挥重要作用,并且具有协同作用。

PTTG和b-FGF在肾癌中的表达

目的探讨垂体肿瘤转化基因(PTTG)和碱性成纤维细胞生长因子(b-FGF)在肾癌中的表达及其与临床病理学参数之间的关系。方法采用免疫组织化学链霉菌抗生物素蛋白过氧化酶(SP)法检测40例肾细胞癌中PTTG和b-FGF的表达情况。结果PTTG、b-FGF在肾癌和正常肾组织中的阳性表达率分别为:52.5%和8.33%(P<0.05),62.5%和16%(P<0.05)。PTTG表达程度与肾癌的临床分期和转移有关(P<0.05),而与其细胞学类型和分级无关(P>0.05)。b-FGF表达程度与肾癌的分级、临床分期和转移有关(P<0.05),而与其细胞学类型无关(P>0.05)。结论PTTG蛋白、b-FGF的高表达参与了肾细胞癌的发展,在肾癌的侵袭和转移中发挥重要作用。

垂体瘤发病相关基因的大规模筛选及鉴定

目的：探讨垂体瘤与垂体组织差异表达基因的大规模筛选及鉴定。方法：垂体无功能腺瘤及正常垂体组织 cDNA文库构建，大规模表达序列标签 (expressed sequence tags， ESTs)测序、表达谱差异的生物信息学处理及半定量逆转录－聚合酶链反应 (reverse transcriptase polymerase chain reaction， RT-PCR)鉴定。结果：从无功能腺瘤和正常垂体组织中分别获得 1253和 7222个质量满意的 ESTs；垂体瘤中已知基因的品种数 200个，经组织表达谱差异比较的统计学软件处理，在这些已知基因中，差异的可靠性≥ 0.99者 38个，≥ 0.95者 130个；其中与细胞生长、分化相关的 G2类基因共 17个； 6个经半定量 RT-PCR证实 4个基因的表达在垂体瘤中明显高于正常垂体。结论：与细胞生长、分化相关的 G2类基因 MEIS2、 SMT3C、 C1D、 BUB3在垂体瘤中表达高于正常垂体，推测可能与垂体瘤的发生有关。