BACKGROUND Cholangiocarcinoma(CCA)represents a rare but highly aggressive malignancy that is often challenging to diagnose,especially in early stages.The role of existing tumor biomarkers for CCA diagnosis,remains con...BACKGROUND Cholangiocarcinoma(CCA)represents a rare but highly aggressive malignancy that is often challenging to diagnose,especially in early stages.The role of existing tumor biomarkers for CCA diagnosis,remains controversial due to their low sensitivity and specificity.Increasing evidence has implicated long non-coding ribonucleic acid polymorphisms with cancer susceptibility in a variety of tumor types.The association between long non-coding ribonucleic acid homeobox protein transcript antisense intergenic ribonucleic acid(HOTAIR)polymorphisms and CCA risk has not been reported yet.AIM To investigate the influence of HOTAIR variants on the risk of CCA development.METHODS We conducted a case-control study in which three HOTAIR single nucleotide polymorphisms(rs920778,rs4759314 and rs7958904)were genotyped in a Greek cohort.Our study population included 122 CCA patients(80 males and 42 females)and 165 healthy controls.The polymorphisms under investigation were examined in peripheral blood samples.RESULTS HOTAIR rs4759314 AG and GG genotypes were associated with a significantly increased CCA risk[P=0.004,odds ratio:3.13;95%confidence interval:1.65-5.91 and P=0.005,odds ratio:12.31;95%confidence interval:1.48-101.87,respectively].However,no significant associations of HOTAIR rs920778,and rs7958904 were detected.Similarly,we found no significant associations between rs4759314 AA genotype and CCA susceptibility.CONCLUSION HOTAIR rs4759314 AG and GG genotypes may be implicated with CCA development and may serve as a potential diagnostic biomarker.展开更多
AIM To identify functional proteins involved in pancreaticduodenal homeobox-1(PDX1)-mediated effects on gastric carcinogenesis.METHODS A PDX1-overexpressed model was established by transfecting gastric cancer cell lin...AIM To identify functional proteins involved in pancreaticduodenal homeobox-1(PDX1)-mediated effects on gastric carcinogenesis.METHODS A PDX1-overexpressed model was established by transfecting gastric cancer cell line SGC7901 with pcDNA3.1(+)-PDX1 vector(SGC-PDX1). Transfection with empty pcDNA3.1 vector(SGC-pcDNA) served as control. Comparative protein profiles of the two groups were analyzed by two-dimensional electrophoresis basedproteomics(2 DE gel-based proteomics). The differential proteins identified by 2 DE were further validated by qRTPCR and immunoblotting. Finally, co-immunoprecipitation was used to determine any direct interactions between PDX1 and the differential proteins.RESULTS2 DE gel proteomics identified seven differential proteins in SGC-PDX1 when compared with those in SGC-pcDNA. These included four heat shock proteins(HSPs; HSP70 p1 B, HSP70 p8, HSP60, HSP27) and three other proteins(ER60, laminin receptor 1, similar to epsilon isoform of 14-3-3 protein). Immunoblotting validated the expression of the HSPs(HSP70, HSP60, HSP27). Furthermore, their expressions were lowered to 80%, 20% and 24%, respectively, in SGC-PDX1, while PDX1 exhibited a 9-fold increase, compared to SGC-pcDNA. However, qRT-PCR analysis revealed that mRNA levels of the HSP s were increased in SGC-PDX1, suggesting that the expression of the HSP s was post-translationally regulated by the PDX1 protein. Finally, co-immunoprecipitation failed to identify any direct interaction between PDX1 and HSP70 proteins.CONCLUSION This study demonstrates the potential involvement of HSPs in PDX1-mediated effects on the genesis of gastric cancer.展开更多
Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) co...Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) constitute a family of transcription factors that are characterized by the presence of a DNA-binding domain closely linked with leucine zipper motif functioning in dimer formation. This type of association is unique to plants and considered as an excellent candidate to activate developmental responses to altering environmental conditions. Cotton is the most important fiber plant with a lot of local and commercial uses in the world. HD-Zip proteins not only have key roles in different stages of vascular and inter-fascicular fiber differentiation of cotton but also are suggested to have an important role against abiotic stress that is one of the key factors limiting cotton productivity. Plants have developed various strategies to manage stress conditions through a combination of metabolic, physiological and morphological adaptations. These adaptive changes rely largely on alterations in gene expression. Therefore, transcriptional regulators play a crucial role in stress tolerance. Being a transcription factor HD-Zip might be a useful target for genetic engineering to generate multiple stress tolerance in susceptible plants. In the following chapter, we discussed how the HD-Zip proteins would play a useful role for cotton development both in fiber production and stress adaptation.展开更多
目的本研究旨在阐明具有放射基因组特征的基于CT图像的活检以预测非小细胞肺癌(NSCLC)患者仅同源域蛋白同源盒(homeodomain-only protein homeobox,HOPX)基因表达状态和预后。方法根据HOPX的表达将患者标记为HOPX阴性或阳性,并将其分为...目的本研究旨在阐明具有放射基因组特征的基于CT图像的活检以预测非小细胞肺癌(NSCLC)患者仅同源域蛋白同源盒(homeodomain-only protein homeobox,HOPX)基因表达状态和预后。方法根据HOPX的表达将患者标记为HOPX阴性或阳性,并将其分为训练数据集(n=92)和测试数据集(n=24)。在对116例患者进行基因与图像特征的相关性分析中,从1218个图像特征中选出了8个与HOPX表达相关的显著特征作为放射基因组特征候选,以预测HOPX的表达状态和预后。结果通过叠加集成学习模型建立具有放射基因组特征的影像活检模型,在测试数据集中,模型显示出对HOPX表达的预测能力,ROC曲线下的面积为0.873,Kaplan-Meier曲线的预测能力(P=0.0066)。结论具有放射基因组特征的基于CT图像的活检可以帮助医生预测HOPX在非小细胞肺癌中的表达状况和预后。展开更多
AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood...AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected.After preparing tissue microarrays and blood smears HGFR,caudal type homeobox 2(CDX2),prominin-1(CD133) and Musashi-1conventional and double fluorescent immunolabelings were performed.Immunostained samples were digitalized using high-resolution Mirax Desk instrument,and analyzed with the Mirax TMA Module software.For semiquantitative counting of immunopositive lamina propria(LP) cells 5 fields of view were counted at magnification x 200 in each sample core,then mean ± SD were determined.In case of peripheral blood smears,30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells(mean ± SD) was determined.Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected.Gene expression analysis of HGFR,CDX2,CD133,leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5),Musashi-1 and cytokeratin20(CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction(RT-PCR).RESULTS:By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR,higher number of HGFR(blood:6.7 ± 1.22 vs 38.5 ±3.18;LP:2.25 ± 0.85 vs 9.22 ± 0.65;P < 0.05),CDX2(blood:0 vs 0.94 ± 0.64;LP:0.75 ± 0.55 vs 2.11± 0.75;P < 0.05),CD133(blood:1.1 ± 0.72 vs 8.3± 1.08;LP:11.1 ± 0.85 vs 26.28 ± 1.71;P < 0.05)and Musashi-1(blood and LP:0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls.HGFR/CDX2(blood:0 vs 1± 0.59;LP:0.8 ± 0.69 vs 2.06 ± 0.72,P < 0.05)and Musashi-1/CDX2(blood and LP:0 vs scattered) coexpressions were found in blood and lamina propria of UC samples.HGFR/CD133 and CD133/CDX2 coexpressions appeared only in UC lamina propria samples.CDX2,Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.CONCLUSION:In active UC,a portion of circulating HGFR-expressing cells are committed to the epithelial lineage,and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.展开更多
目的:探讨益胃消瘀颗粒治疗萎缩性胃炎胃黏膜肠化的治疗作用与机制。方法:8周龄雄性SD大鼠60只,按随机数字表法分为空白组、模型组、胃复春组、益胃消瘀颗粒高剂量组、益胃消瘀颗粒中剂量组、益胃消瘀颗粒低剂量组共6组,N-甲基-N’-硝基...目的:探讨益胃消瘀颗粒治疗萎缩性胃炎胃黏膜肠化的治疗作用与机制。方法:8周龄雄性SD大鼠60只,按随机数字表法分为空白组、模型组、胃复春组、益胃消瘀颗粒高剂量组、益胃消瘀颗粒中剂量组、益胃消瘀颗粒低剂量组共6组,N-甲基-N’-硝基-N-亚硝基胍(MNNG)和法莫替丁联合造模。6个月及7个月后分次抽检3只,造模成功后开始进入实验给药阶段,给药3个月后观察大鼠胃黏膜组织学改变,RT-qPCR检测胃黏膜标本中尾型同源盒转录因子2(caudal-related homeobox transcription factor2,CDX2)、干细胞转录调控因子性别决定相关基因簇2(Sex de-termining region Y-box 2,SOX2)的表达,Western blot检测胃黏膜中NK6同源框蛋白3(NK6 Homeobox Protein 3,NKX6.3)、骨形成蛋白4(bone morphogenic protein 4,BMP4)的表达。结果:光镜下观察胃黏膜病理组织,模型组可见黏膜萎缩及肠化,益胃消瘀颗粒高、中剂量组及胃复春组萎缩及肠化均较模型组明显改善,低剂量组萎缩及肠化有所减轻;与空白组比较,模型组NKX6.3、Sox2表达明显下降,BMP4、CDx2表达明显升高;与模型组比较,益胃消瘀颗粒高、中剂量组和胃复春组NKX6.3表达明显上升,益胃消瘀颗粒高、中、低剂量组及胃复春组Sox2表达明显升高,BMP4、CDx2表达明显下降。结论:益胃消瘀颗粒能够改善萎缩性胃炎肠化大鼠胃黏膜组织病理形态,其机制可能与激活NKX6.3、下调Cdx2、上调Sox2和抑制BMP4表达有关。展开更多
基金The Hellenic Society of Medical Oncology,No.8021/25.09.2020.
文摘BACKGROUND Cholangiocarcinoma(CCA)represents a rare but highly aggressive malignancy that is often challenging to diagnose,especially in early stages.The role of existing tumor biomarkers for CCA diagnosis,remains controversial due to their low sensitivity and specificity.Increasing evidence has implicated long non-coding ribonucleic acid polymorphisms with cancer susceptibility in a variety of tumor types.The association between long non-coding ribonucleic acid homeobox protein transcript antisense intergenic ribonucleic acid(HOTAIR)polymorphisms and CCA risk has not been reported yet.AIM To investigate the influence of HOTAIR variants on the risk of CCA development.METHODS We conducted a case-control study in which three HOTAIR single nucleotide polymorphisms(rs920778,rs4759314 and rs7958904)were genotyped in a Greek cohort.Our study population included 122 CCA patients(80 males and 42 females)and 165 healthy controls.The polymorphisms under investigation were examined in peripheral blood samples.RESULTS HOTAIR rs4759314 AG and GG genotypes were associated with a significantly increased CCA risk[P=0.004,odds ratio:3.13;95%confidence interval:1.65-5.91 and P=0.005,odds ratio:12.31;95%confidence interval:1.48-101.87,respectively].However,no significant associations of HOTAIR rs920778,and rs7958904 were detected.Similarly,we found no significant associations between rs4759314 AA genotype and CCA susceptibility.CONCLUSION HOTAIR rs4759314 AG and GG genotypes may be implicated with CCA development and may serve as a potential diagnostic biomarker.
基金Supported by Guangdong Natural Science Fund,No.2016A030313765Guangdong medical scientific research fund,No.A2017070 and No.A2017122
文摘AIM To identify functional proteins involved in pancreaticduodenal homeobox-1(PDX1)-mediated effects on gastric carcinogenesis.METHODS A PDX1-overexpressed model was established by transfecting gastric cancer cell line SGC7901 with pcDNA3.1(+)-PDX1 vector(SGC-PDX1). Transfection with empty pcDNA3.1 vector(SGC-pcDNA) served as control. Comparative protein profiles of the two groups were analyzed by two-dimensional electrophoresis basedproteomics(2 DE gel-based proteomics). The differential proteins identified by 2 DE were further validated by qRTPCR and immunoblotting. Finally, co-immunoprecipitation was used to determine any direct interactions between PDX1 and the differential proteins.RESULTS2 DE gel proteomics identified seven differential proteins in SGC-PDX1 when compared with those in SGC-pcDNA. These included four heat shock proteins(HSPs; HSP70 p1 B, HSP70 p8, HSP60, HSP27) and three other proteins(ER60, laminin receptor 1, similar to epsilon isoform of 14-3-3 protein). Immunoblotting validated the expression of the HSPs(HSP70, HSP60, HSP27). Furthermore, their expressions were lowered to 80%, 20% and 24%, respectively, in SGC-PDX1, while PDX1 exhibited a 9-fold increase, compared to SGC-pcDNA. However, qRT-PCR analysis revealed that mRNA levels of the HSP s were increased in SGC-PDX1, suggesting that the expression of the HSP s was post-translationally regulated by the PDX1 protein. Finally, co-immunoprecipitation failed to identify any direct interaction between PDX1 and HSP70 proteins.CONCLUSION This study demonstrates the potential involvement of HSPs in PDX1-mediated effects on the genesis of gastric cancer.
文摘Transcription factors play key roles in plant development and stress responses through their interaction with cis-elements and/or other transcription factors. Homeodomain associated leucine zipper proteins (HD-Zip) constitute a family of transcription factors that are characterized by the presence of a DNA-binding domain closely linked with leucine zipper motif functioning in dimer formation. This type of association is unique to plants and considered as an excellent candidate to activate developmental responses to altering environmental conditions. Cotton is the most important fiber plant with a lot of local and commercial uses in the world. HD-Zip proteins not only have key roles in different stages of vascular and inter-fascicular fiber differentiation of cotton but also are suggested to have an important role against abiotic stress that is one of the key factors limiting cotton productivity. Plants have developed various strategies to manage stress conditions through a combination of metabolic, physiological and morphological adaptations. These adaptive changes rely largely on alterations in gene expression. Therefore, transcriptional regulators play a crucial role in stress tolerance. Being a transcription factor HD-Zip might be a useful target for genetic engineering to generate multiple stress tolerance in susceptible plants. In the following chapter, we discussed how the HD-Zip proteins would play a useful role for cotton development both in fiber production and stress adaptation.
文摘目的本研究旨在阐明具有放射基因组特征的基于CT图像的活检以预测非小细胞肺癌(NSCLC)患者仅同源域蛋白同源盒(homeodomain-only protein homeobox,HOPX)基因表达状态和预后。方法根据HOPX的表达将患者标记为HOPX阴性或阳性,并将其分为训练数据集(n=92)和测试数据集(n=24)。在对116例患者进行基因与图像特征的相关性分析中,从1218个图像特征中选出了8个与HOPX表达相关的显著特征作为放射基因组特征候选,以预测HOPX的表达状态和预后。结果通过叠加集成学习模型建立具有放射基因组特征的影像活检模型,在测试数据集中,模型显示出对HOPX表达的预测能力,ROC曲线下的面积为0.873,Kaplan-Meier曲线的预测能力(P=0.0066)。结论具有放射基因组特征的基于CT图像的活检可以帮助医生预测HOPX在非小细胞肺癌中的表达状况和预后。
基金Cell Analysis Laboratory, 2nd Department of Internal Medicine, and the 1st Department of Pathology and Experimental Oncology, Semmelweis University for their technical support
文摘AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected.After preparing tissue microarrays and blood smears HGFR,caudal type homeobox 2(CDX2),prominin-1(CD133) and Musashi-1conventional and double fluorescent immunolabelings were performed.Immunostained samples were digitalized using high-resolution Mirax Desk instrument,and analyzed with the Mirax TMA Module software.For semiquantitative counting of immunopositive lamina propria(LP) cells 5 fields of view were counted at magnification x 200 in each sample core,then mean ± SD were determined.In case of peripheral blood smears,30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells(mean ± SD) was determined.Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected.Gene expression analysis of HGFR,CDX2,CD133,leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5),Musashi-1 and cytokeratin20(CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction(RT-PCR).RESULTS:By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR,higher number of HGFR(blood:6.7 ± 1.22 vs 38.5 ±3.18;LP:2.25 ± 0.85 vs 9.22 ± 0.65;P < 0.05),CDX2(blood:0 vs 0.94 ± 0.64;LP:0.75 ± 0.55 vs 2.11± 0.75;P < 0.05),CD133(blood:1.1 ± 0.72 vs 8.3± 1.08;LP:11.1 ± 0.85 vs 26.28 ± 1.71;P < 0.05)and Musashi-1(blood and LP:0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls.HGFR/CDX2(blood:0 vs 1± 0.59;LP:0.8 ± 0.69 vs 2.06 ± 0.72,P < 0.05)and Musashi-1/CDX2(blood and LP:0 vs scattered) coexpressions were found in blood and lamina propria of UC samples.HGFR/CD133 and CD133/CDX2 coexpressions appeared only in UC lamina propria samples.CDX2,Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.CONCLUSION:In active UC,a portion of circulating HGFR-expressing cells are committed to the epithelial lineage,and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.
文摘目的:探讨益胃消瘀颗粒治疗萎缩性胃炎胃黏膜肠化的治疗作用与机制。方法:8周龄雄性SD大鼠60只,按随机数字表法分为空白组、模型组、胃复春组、益胃消瘀颗粒高剂量组、益胃消瘀颗粒中剂量组、益胃消瘀颗粒低剂量组共6组,N-甲基-N’-硝基-N-亚硝基胍(MNNG)和法莫替丁联合造模。6个月及7个月后分次抽检3只,造模成功后开始进入实验给药阶段,给药3个月后观察大鼠胃黏膜组织学改变,RT-qPCR检测胃黏膜标本中尾型同源盒转录因子2(caudal-related homeobox transcription factor2,CDX2)、干细胞转录调控因子性别决定相关基因簇2(Sex de-termining region Y-box 2,SOX2)的表达,Western blot检测胃黏膜中NK6同源框蛋白3(NK6 Homeobox Protein 3,NKX6.3)、骨形成蛋白4(bone morphogenic protein 4,BMP4)的表达。结果:光镜下观察胃黏膜病理组织,模型组可见黏膜萎缩及肠化,益胃消瘀颗粒高、中剂量组及胃复春组萎缩及肠化均较模型组明显改善,低剂量组萎缩及肠化有所减轻;与空白组比较,模型组NKX6.3、Sox2表达明显下降,BMP4、CDx2表达明显升高;与模型组比较,益胃消瘀颗粒高、中剂量组和胃复春组NKX6.3表达明显上升,益胃消瘀颗粒高、中、低剂量组及胃复春组Sox2表达明显升高,BMP4、CDx2表达明显下降。结论:益胃消瘀颗粒能够改善萎缩性胃炎肠化大鼠胃黏膜组织病理形态,其机制可能与激活NKX6.3、下调Cdx2、上调Sox2和抑制BMP4表达有关。
