The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epith...The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.展开更多
BACKGROUND Thrombocytopenia 2,an autosomal dominant inherited disease characterized by moderate thrombocytopenia,predisposition to myeloid malignancies and normal platelet size and function,can be caused by 5’-untran...BACKGROUND Thrombocytopenia 2,an autosomal dominant inherited disease characterized by moderate thrombocytopenia,predisposition to myeloid malignancies and normal platelet size and function,can be caused by 5’-untranslated region(UTR)point mutations in ankyrin repeat domain containing 26(ANKRD26).Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1)have been identified as negative regulators of ANKRD26.However,the positive regulators of ANKRD26 are still unknown.AIM To prove the positive regulatory effect of GATA binding protein 2(GATA2)on ANKRD26 transcription.METHODS Human induced pluripotent stem cells derived from bone marrow(hiPSC-BM)INTRODUCTION Ankyrin repeat domain containing protein 26(ANKRD26)acts as a regulator of adipogenesis and is involved in the regulation of feeding behavior[1-3].The ANKRD26 gene is located on chromosome 10 and shares regions of homology with the primate-specific gene family POTE.According to the Human Protein Atlas database,the ANKRD26 protein is localized to the Golgi apparatus and vesicles,and its expression can be detected in nearly all human tissues[4].Moreover,UniProt annotation revealed that ANKRD26 is localized in the centrosome and contains coiled-coil domains formed by spectrin helices and ankyrin repeats[5,6].The most common disease related to ANKRD26 is thrombocytopenia 2(THC2),which is a rare autosomal dominant inherited disease characterized by lifelong mild-to-moderate thrombocytopenia and mild bleeding[7-9].Caused by the variants in the 5’-untranslated region(UTR)of ANKRD26,THC2 is defined by a decrease in the number of platelets in circulating blood and results in increased bleeding and decreased clotting ability[8,10].Due to the point mutations that occur in the 5’-UTR of ANKRD26,its negative transcription factors(TFs),Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1),lose their repression effect[11].The persistent expression of ANKRD26 increases the activity of the mitogen activated protein kinase and extracellular signal regulated kinase 1/2 signaling pathways,which are potentially involved in the regulation of thrombopoietin-dependent signaling and further impair proplatelet formation by megakaryocytes(MKs)[11].However,the positive regulators of ANKRD26,which might be associated with THC2 pathology,are still unknown.展开更多
Background:Clear cell renal cell carcinoma(ccRCC)stands as the most prevalent form of kidney cancer,accounting for a significant proportion of malignancies affecting the kidneys.ccRCC is well known as a type of tumour...Background:Clear cell renal cell carcinoma(ccRCC)stands as the most prevalent form of kidney cancer,accounting for a significant proportion of malignancies affecting the kidneys.ccRCC is well known as a type of tumour with immunogenicity.Immune checkpoint inhibitors(ICIs)aim to enhance the anticancer immune response in ccRCC by blocking programmed cell death 1 ligand 1/programmed death 1(PD-L1/PD-1)pathways.In a previous study,we showed that RING finger protein 26(RNF26)degrades chromobox 7(CBX7)to activate the tumor necrosis factor(TNF)in ccRCC.Methods:We analyzed The Cancer Genome Atlas(TCGA)database using the R package ESTIMATE and found that RNF26 was significantly associated with ccRCC immune infiltration.The relationship between RNF26 and the PD1 checkpoint signaling pathway was detected by enrichment analysis.In addition,the molecular mechanism of RNF26 up-regulation of PD-L1 was detected by transcriptome sequencing,RT-qPCR,and Western Blot in ccRCC cell lineages 786-O and A498 cells.The transplantation tumor experiments in C57BL/6 mice were used to test the efficacy of anti-PD1 and knockdown of RNF26 in vivo.Results:We showed that RNF26 suppressed the immune response to ccRCC.Next,we revealed that RNF26 activated the PD-1 checkpoint pathway to suppress the immune response to ccRCC,possibly via the CBX7/PD-L1 axis.Conclusion:The suggestion derived from our results is that targeting RNF26 holds the potential to amplify the efficacy of anticancer immunotherapies in the treatment of ccRCC.展开更多
Objective： The aim of the study was to compare efficacies and safeties of 2 different treatments of whole brain radiotherapy （WBRT） sequential or concomitant Vm26/DDP for small cell lung cancer （SCLC） patients wi...Objective： The aim of the study was to compare efficacies and safeties of 2 different treatments of whole brain radiotherapy （WBRT） sequential or concomitant Vm26/DDP for small cell lung cancer （SCLC） patients with brain metastases. Methods： A total of 39 patients were randomly divided into sequential chemoradiotherapy regime （A group, 20 patients） and concomitant chemoradiotherapy regime （B group, 19 patients）. The close of WBRT was 36 Gy in 18-20 fractions, chemotherapy of Vm26/DDP regimen with teniposide 60 mg/m^2 on dl to d3 and cisplatin 20 mg/m^2 on dl to d5, repeating every 3 weeks. The response was evaluated after WBRT and 2 cycles of chemotherapy. Results： Total response rates of A and B groups were 70.0% and 78.9% respectively （P = 0.520）. The median survival was 11 months in A group and 10 months in B group. Six, twelve and eighteen months cumulative survival rates of A and B groups were 75.0%, 42.5%, 26.2%, and 81.6%, 26.4%, 10.5%, respectively （χ^2 = 0.383, P 〉 0.05）. Response rate and the number of brain metastases were independent prognostic factors. Conclusion： Both sequential and concomitant chemoradiotherapy groups are effective, and the main toxicity with myelosuppression is tolerable after therapy. It can be applied firstly and effectively to the SCLC patients with brain metastases in clinic.展开更多
Tissue engineering techniques for cartilage re-pair to heal defects in joint surfaces is a clinical practice. Harvested autologous chondrocytes are expanded in culture and delivered in a suitable carrier medium back i...Tissue engineering techniques for cartilage re-pair to heal defects in joint surfaces is a clinical practice. Harvested autologous chondrocytes are expanded in culture and delivered in a suitable carrier medium back into the patient>s joint de-fect. The defect is then subsequently filled by new cartilage. Whether the cells in the repair tissue originate from the engineered tissue of the host or are derived from the surrounding original cartilage remains a relevant question for the ap-plied therapy. To answer this several methods exist to track cells, such as transfection of cells with LacZ carrying viruses, radio labeling with 111 IN or 51 Cr or fluorescent labeling with FDA. However, these techniques have drawbacks such as they may influence cellular properties, are radioactive and or quickly lose their tracking ability. New fluorescent probes are easier to handle and do not to interfere with cells. PKH 26劌 is a relatively new cell-labeling agent, but few data exist on the application of this dye in chondrocytes in vitro and in vivo. 5-chloromethylfluorescein diacetate - CMFDA (¨cell tracker green〔) is an established fluores-cent probe for imaging the dynamic processes of cell proliferation in vitro and in vivo. Likewise, several studies exist on different cell types. However, little data are available for chondro-cytes. The first aim of the study was to evaluate qualitative differences in fluorescence pattern after labeling of articular, auricular and costal chondrocytes. Secondly, we evaluated the influ-ence of labeling with CMFDA on cellular adhe-sion properties. The third aim was to compare the duration of cell labeling of chondrocytes of different origin with established CMFDA as stan-dard and PKH 26潴 for 3 cell generations in vitro and 12 weeks in vivo. We show that chondro-cytes from different origin can be labeled effec-tively with both PKH 26潴 and CMFDA. The PKH 26潴 labeled articular chondrocytes maintained fluorescence longer than CMFDA in vitro and in vivo. A higher percentage of articular chondro-cytes remained stained at 63 days than auricular or costal chondrocytes.展开更多
Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-sp...Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-special 5(GAS5)on proliferation and apoptosis of hepatocellular carcinoma(HCC)cells through miR-26a-5p action.Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC patients by RT-qPCR.The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,which was also highly expressed in HCC tissue.The relationship between GAS5 and miR-26a-5p was confirmed using a luciferase reporter assay.The role of these lncRNAs was further explored by transfecting plasmids into SMMC-7721 cells and classifying the cells as follows:NC group,GAS5 group,anti-miR-26a-5p group,and GAS5+miR-26a-5p group.Cell proliferation,cell cycle,and apoptosis were detected in each group.The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN)was analyzed by TargetScan database prediction and luciferase reporter assay.Western blotting was used to quantify PTEN,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),cyclin D1,and human P27 protein(P27).Results GAS5 was downregulated,while miR-26a-5p was upregulated in HCC tissue compared to in paracancerous tissue.High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation,increased the number of G0/G1 phase cells,promoted cell apoptosis,promoted PTEN and P27 expression,and inhibited PI3K,P-Akt,and cyclin D1 expression at the protein level.Upregulation of miR-26a-5p attenuated the effects of GAS5 upregulation on the proliferation,cell cycle,and apoptosis of HCC cells and on the expression of PTNE/PI3K/Akt signaling pathway-related proteins.Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/Akt signaling pathway and are linked to upregulation of miR-26a-5p.展开更多
基金supported by the China Postdoctoral Science Foundation(2019M653776 and 2020M673516)the Natural Science Basis Research Plan in Shaanxi Province of China(2023-JC-QN-0181)+1 种基金the Shaanxi Livestock and Poultry Breeding Double-chain Fusion Key Project,China(2022GD-TSLD-46-0202)the Natural Science Fundation of Tibet Autonomous Region,China(XZ202101ZR0063G)。
文摘The development of receptive endometrium(RE) from pre-receptive endometrium(PE) for successful embryo implantation is a complex dynamic process in which the morphology and physiological states of the endometrial epithelium undergo a series of significant changes, including cell proliferation and apoptosis. However, the molecular mechanisms are not yet fully understood. In this study, a higher circRNA3669 level was observed in PE than in RE of goats. Functional assays revealed that this overexpression promoted the proliferation of goat endometrial epithelial cells(GEECs) by activating the expression of genes related to the PI3K/AKT-mTOR and MAPK pathways,thereby inhibiting apoptosis in vitro. Furthermore, circRNA3669 functioned as a competing endogenous RNA(ceRNA) to upregulate Reticulocalbin-2(RCN2) expression at the post-transcriptional level by interacting with and downregulating miR-26a in GEECs. In addition, RCN2, which is highly expressed in the PE of goats, was found to be regulated by β-estradiol(E2) and progesterone(P4). Our results demonstrated that RCN2 also affected the key proteins PI3K, AKT, mTOR, JNK, and P38 in the PI3K/AKT-mTOR and MAPK pathways, thereby facilitating GEECs proliferation and suppressing their apoptosis in vitro. Collectively, we constructed a new circRNA3669-miR-26aRCN2 regulatory network in GEECs, which further provides strong evidence that circRNA could potentially play a crucial regulatory role in the development of RE in goats.
基金Supported by General Program of National Natural Science Foundation of China,No.81770197Scientific and Technological Research Major Program of Chongqing Municipal Education Commission,No.KJZD-M202312802+1 种基金Chongqing Natural Science Foundation of China,No.CSTB2022NSCQ-MSX0190,No.CSTB2022NSCQ-MSX0176,and No.cstc2020jcyj-msxmX0051Xinqiao Young Postdoc Talent Incubation Program,No.2022YQB098.
文摘BACKGROUND Thrombocytopenia 2,an autosomal dominant inherited disease characterized by moderate thrombocytopenia,predisposition to myeloid malignancies and normal platelet size and function,can be caused by 5’-untranslated region(UTR)point mutations in ankyrin repeat domain containing 26(ANKRD26).Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1)have been identified as negative regulators of ANKRD26.However,the positive regulators of ANKRD26 are still unknown.AIM To prove the positive regulatory effect of GATA binding protein 2(GATA2)on ANKRD26 transcription.METHODS Human induced pluripotent stem cells derived from bone marrow(hiPSC-BM)INTRODUCTION Ankyrin repeat domain containing protein 26(ANKRD26)acts as a regulator of adipogenesis and is involved in the regulation of feeding behavior[1-3].The ANKRD26 gene is located on chromosome 10 and shares regions of homology with the primate-specific gene family POTE.According to the Human Protein Atlas database,the ANKRD26 protein is localized to the Golgi apparatus and vesicles,and its expression can be detected in nearly all human tissues[4].Moreover,UniProt annotation revealed that ANKRD26 is localized in the centrosome and contains coiled-coil domains formed by spectrin helices and ankyrin repeats[5,6].The most common disease related to ANKRD26 is thrombocytopenia 2(THC2),which is a rare autosomal dominant inherited disease characterized by lifelong mild-to-moderate thrombocytopenia and mild bleeding[7-9].Caused by the variants in the 5’-untranslated region(UTR)of ANKRD26,THC2 is defined by a decrease in the number of platelets in circulating blood and results in increased bleeding and decreased clotting ability[8,10].Due to the point mutations that occur in the 5’-UTR of ANKRD26,its negative transcription factors(TFs),Runt related transcription factor 1(RUNX1)and friend leukemia integration 1(FLI1),lose their repression effect[11].The persistent expression of ANKRD26 increases the activity of the mitogen activated protein kinase and extracellular signal regulated kinase 1/2 signaling pathways,which are potentially involved in the regulation of thrombopoietin-dependent signaling and further impair proplatelet formation by megakaryocytes(MKs)[11].However,the positive regulators of ANKRD26,which might be associated with THC2 pathology,are still unknown.
基金supported by Natural Science Foundation of Hunan Province of China(Grant No.2022JJ30870(Liang Zhu)).
文摘Background:Clear cell renal cell carcinoma(ccRCC)stands as the most prevalent form of kidney cancer,accounting for a significant proportion of malignancies affecting the kidneys.ccRCC is well known as a type of tumour with immunogenicity.Immune checkpoint inhibitors(ICIs)aim to enhance the anticancer immune response in ccRCC by blocking programmed cell death 1 ligand 1/programmed death 1(PD-L1/PD-1)pathways.In a previous study,we showed that RING finger protein 26(RNF26)degrades chromobox 7(CBX7)to activate the tumor necrosis factor(TNF)in ccRCC.Methods:We analyzed The Cancer Genome Atlas(TCGA)database using the R package ESTIMATE and found that RNF26 was significantly associated with ccRCC immune infiltration.The relationship between RNF26 and the PD1 checkpoint signaling pathway was detected by enrichment analysis.In addition,the molecular mechanism of RNF26 up-regulation of PD-L1 was detected by transcriptome sequencing,RT-qPCR,and Western Blot in ccRCC cell lineages 786-O and A498 cells.The transplantation tumor experiments in C57BL/6 mice were used to test the efficacy of anti-PD1 and knockdown of RNF26 in vivo.Results:We showed that RNF26 suppressed the immune response to ccRCC.Next,we revealed that RNF26 activated the PD-1 checkpoint pathway to suppress the immune response to ccRCC,possibly via the CBX7/PD-L1 axis.Conclusion:The suggestion derived from our results is that targeting RNF26 holds the potential to amplify the efficacy of anticancer immunotherapies in the treatment of ccRCC.
文摘Objective： The aim of the study was to compare efficacies and safeties of 2 different treatments of whole brain radiotherapy （WBRT） sequential or concomitant Vm26/DDP for small cell lung cancer （SCLC） patients with brain metastases. Methods： A total of 39 patients were randomly divided into sequential chemoradiotherapy regime （A group, 20 patients） and concomitant chemoradiotherapy regime （B group, 19 patients）. The close of WBRT was 36 Gy in 18-20 fractions, chemotherapy of Vm26/DDP regimen with teniposide 60 mg/m^2 on dl to d3 and cisplatin 20 mg/m^2 on dl to d5, repeating every 3 weeks. The response was evaluated after WBRT and 2 cycles of chemotherapy. Results： Total response rates of A and B groups were 70.0% and 78.9% respectively （P = 0.520）. The median survival was 11 months in A group and 10 months in B group. Six, twelve and eighteen months cumulative survival rates of A and B groups were 75.0%, 42.5%, 26.2%, and 81.6%, 26.4%, 10.5%, respectively （χ^2 = 0.383, P 〉 0.05）. Response rate and the number of brain metastases were independent prognostic factors. Conclusion： Both sequential and concomitant chemoradiotherapy groups are effective, and the main toxicity with myelosuppression is tolerable after therapy. It can be applied firstly and effectively to the SCLC patients with brain metastases in clinic.
文摘Tissue engineering techniques for cartilage re-pair to heal defects in joint surfaces is a clinical practice. Harvested autologous chondrocytes are expanded in culture and delivered in a suitable carrier medium back into the patient>s joint de-fect. The defect is then subsequently filled by new cartilage. Whether the cells in the repair tissue originate from the engineered tissue of the host or are derived from the surrounding original cartilage remains a relevant question for the ap-plied therapy. To answer this several methods exist to track cells, such as transfection of cells with LacZ carrying viruses, radio labeling with 111 IN or 51 Cr or fluorescent labeling with FDA. However, these techniques have drawbacks such as they may influence cellular properties, are radioactive and or quickly lose their tracking ability. New fluorescent probes are easier to handle and do not to interfere with cells. PKH 26劌 is a relatively new cell-labeling agent, but few data exist on the application of this dye in chondrocytes in vitro and in vivo. 5-chloromethylfluorescein diacetate - CMFDA (¨cell tracker green〔) is an established fluores-cent probe for imaging the dynamic processes of cell proliferation in vitro and in vivo. Likewise, several studies exist on different cell types. However, little data are available for chondro-cytes. The first aim of the study was to evaluate qualitative differences in fluorescence pattern after labeling of articular, auricular and costal chondrocytes. Secondly, we evaluated the influ-ence of labeling with CMFDA on cellular adhe-sion properties. The third aim was to compare the duration of cell labeling of chondrocytes of different origin with established CMFDA as stan-dard and PKH 26潴 for 3 cell generations in vitro and 12 weeks in vivo. We show that chondro-cytes from different origin can be labeled effec-tively with both PKH 26潴 and CMFDA. The PKH 26潴 labeled articular chondrocytes maintained fluorescence longer than CMFDA in vitro and in vivo. A higher percentage of articular chondro-cytes remained stained at 63 days than auricular or costal chondrocytes.
文摘Objective Long non-coding RNAs(lncRNAs)regulate tumor development and progression by promoting tumor proliferation,invasion,and metastasis.The aim of the study was to investigate the effects of lncRNA growth arrest-special 5(GAS5)on proliferation and apoptosis of hepatocellular carcinoma(HCC)cells through miR-26a-5p action.Methods Expression levels of GAS5 were detected in cancerous and paracancerous tissue of 80 HCC patients by RT-qPCR.The starBase tool predicted that GAS5 had binding sites for the miRNA miR-26a-5p,which was also highly expressed in HCC tissue.The relationship between GAS5 and miR-26a-5p was confirmed using a luciferase reporter assay.The role of these lncRNAs was further explored by transfecting plasmids into SMMC-7721 cells and classifying the cells as follows:NC group,GAS5 group,anti-miR-26a-5p group,and GAS5+miR-26a-5p group.Cell proliferation,cell cycle,and apoptosis were detected in each group.The relationship between miR-26a-5p and phosphatase and tensin homolog deleted on chromosome 10(PTEN)was analyzed by TargetScan database prediction and luciferase reporter assay.Western blotting was used to quantify PTEN,phosphatidylinositol 3-kinase(PI3K),phosphorylated protein kinase B(p-Akt),cyclin D1,and human P27 protein(P27).Results GAS5 was downregulated,while miR-26a-5p was upregulated in HCC tissue compared to in paracancerous tissue.High GAS5 levels and low miR-26a-5p levels inhibited cell proliferation,increased the number of G0/G1 phase cells,promoted cell apoptosis,promoted PTEN and P27 expression,and inhibited PI3K,P-Akt,and cyclin D1 expression at the protein level.Upregulation of miR-26a-5p attenuated the effects of GAS5 upregulation on the proliferation,cell cycle,and apoptosis of HCC cells and on the expression of PTNE/PI3K/Akt signaling pathway-related proteins.Conclusion Low GAS5 levels regulate the proliferation and apoptosis of HCC cells via the PTNE/PI3K/Akt signaling pathway and are linked to upregulation of miR-26a-5p.
