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Objective To investigate the expression level and clinical significance of serum long noncoding RNA(lncRNA)H19,CHAST in patients with coronary heart disease(CHD).Methods A total of 135 patients with CHD who were admitted to the Department of Cardiovascular Medicine of Lianyungang Second People's Hospital from June 2021 to May 2022 were selected,including 76 cases of a-cute coronary syndrome and 59 cases of chronic coronary syndrome.We selected 62 patients in the control group who were hospitalized and excluded from CHD by coronary angiography.Real-time fluorescent quantitative PCR(RT-PCR)was used to detect the relative expres-sion level of lncRNA H19 and lncRNA CHAST in the circulating serum of the subjects.Meanwhile,the high sensitivity C-reactive pro-tein(hs-CRP)、tumor necrosis factor-α(TNF-α)、interleukin-6(IL-6)、low density lipoprotein(LDL)、high density lipoprotein(HDL)and other related indicators of each group were collected.The receiver operating characteristic(ROC)curve was used to analyze the clinical diagnostic value of circulating serum lncRNA H19 and lncRNA CHAST expression level in the diagnosis of CHD.Pearson cor-relation coefficient was used to analyze the correlation between serum lncRNA H19,lncRNA CHAST level,and other related indicators.Multivariate Logistic regression was used to analyze the risk factors of CHD.Results The serum expression levels of lncRNA H19 and ln-cRNA CHAST in the CHD group were higher than those in the control group(P<0.05).The serum levels of hs-CRP、TNF-α、IL-6、and LDL in the experimental group were higher than those in the control group(P<0.05),and the serum levels of HDL were lower than those in the control group(P<0.05).The expression levels of lncRNA H19 and lncRNA CHAST were positively correlated with hs-CRP,IL-6 and LDL,and were negatively correlated with HDL.lncRNA H19 and lncRNA CHAST are risk factors for CHD,which may have a higher value in the diagnosis of CHD.Conclusion lncRNA H19 and lncRNA CHAST are risk factors for CHD and can be used as serum indicators for the diagnosis of CHD,and the combination of the two has a higher value in the diagnosis of CHD.In addition,the ex-pression levels of both were correlated with the levels of inflammatory factors and apolipoproteins.
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Long non-coding RNA(lncRNA)is a type of non-protein-coding RNA with a length of at least 200 nucleotides.Ex-isting evidence shows that lncRNA affects the growth and development of humans and the occurrence of diseases through various mech-anisms,and plays an irreplaceable role in the differentiation and activation of immune cells and the development process of autoim-mune diseases.In recent years,lncRNA H19 has been found to play a unique regulatory function in pathological processes related to autoimmune diseases,such as inflammation and fibrosis.Therefore,the molecular mechanism involved in H19 is expected to become a potential target for the treatment of autoimmune diseases.In this review,we discussed the related mechanisms of H19 involved in the pathogenesis of autoimmune diseases based on the known functions of H19 and summarized the research results of H19 in several com-mon autoimmune diseases.
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OBJECTIVE@#Hepatic fibrosis has been widely considered as a conjoint consequence of almost all chronic liver diseases. Chuanxiong Rhizoma (Chuanxiong in Chinese, CX) is a traditional Chinese herbal product to prevent cerebrovascular, gynecologic and hepatic diseases. Our previous study found that CX extracts significantly reduced collagen contraction force of hepatic stellate cells (HSCs). Here, this study aimed to compare the protection of different CX extracts on bile duct ligation (BDL)-induced liver fibrosis and investigate plausible underlying mechanisms.@*METHODS@#The active compounds of CX extracts were identified by high performance liquid chromatography (HPLC). Network pharmacology was used to determine potential targets of CX against hepatic fibrosis. Bile duct hyperplasia and liver fibrosis were evaluated by serologic testing and histopathological evaluation. The expression of targets of interest was determined by quantitative real-time PCR (qPCR) and Western blot.@*RESULTS@#Different CX extracts were identified by tetramethylpyrazine, ferulic acid and senkyunolide A. Based on the network pharmacological analysis, 42 overlap targets were obtained via merging the candidates targets of CX and liver fibrosis. Different aqueous, alkaloid and phthalide extracts of CX (CXAE, CXAL and CXPHL) significantly inhibited diffuse severe bile duct hyperplasia and thus suppressed hepatic fibrosis by decreasing CCCTC binding factor (CTCF)-c-MYC-long non-coding RNA H19 (H19) pathway in the BDL-induced mouse model. Meanwhile, CX extracts, especially CXAL and CXPHL also suppressed CTCF-c-MYC-H19 pathway and inhibited ductular reaction in cholangiocytes stimulated with taurocholate acid (TCA), lithocholic acid (LCA) and transforming growth factor beta (TGF-β), as illustrated by decreased bile duct proliferation markers.@*CONCLUSION@#Our data supported that different CX extracts, especially CXAL and CXPHL significantly alleviated hepatic fibrosis and bile duct hyperplasia via inhibiting CTCF-c-MYC-H19 pathway, providing novel insights into the anti-fibrotic mechanism of CX.
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Liver fibrosis is a dynamic wound-healing response characterized by the agglutination of the extracellular matrix (ECM). Si-Wu-Tang (SWT), a traditional Chinese medicine (TCM) formula, is known for treating gynecological diseases and liver fibrosis. Our previous studies demonstrated that long non-coding RNA H19 (H19) was markedly upregulated in fibrotic livers while its deficiency markedly reversed fibrogenesis. However, the mechanisms by which SWT influences H19 remain unclear. Thus, we established a bile duct ligation (BDL)-induced liver fibrosis model to evaluate the hepatoprotective effects of SWT on various cells in the liver. Our results showed that SWT markedly improved ECM deposition and bile duct reactions in the liver. Notably, SWT relieved liver fibrosis by regulating the transcription of genes involved in the cytoskeleton remodeling, primarily in hepatic stellate cells (HSCs), and influencing cytoskeleton-related angiogenesis and hepatocellular injury. This modulation collectively led to reduced ECM deposition. Through extensive bioinformatics analyses, we determined that H19 acted as a miRNA sponge and mainly inhibited miR-200, miR-211, and let7b, thereby regulating the above cellular regulatory pathways. Meanwhile, SWT reversed H19-related miRNAs and signaling pathways, diminishing ECM deposition and liver fibrosis. However, these protective effects of SWT were diminished with the overexpression of H19 in vivo. In conclusion, our study elucidates the underlying mechanisms of SWT from the perspective of H19-related signal networks and proposes a potential SWT-based therapeutic strategy for the treatment of liver fibrosis.
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Humans , RNA, Long Noncoding/genetics , Liver Cirrhosis/genetics , Liver/metabolism , Hepatic Stellate Cells/pathology , MicroRNAs/metabolism , Extracellular Matrix/metabolism , Drugs, Chinese HerbalABSTRACT
Aim To investigate the effect of heroin use in male rats of F0 generation on heroin addiction and relapse in rats of Fl generation and the underlying mechanism. Methods Male rats of F0 generation were treated with different doses of heroin (1, 3, 9 mg • kg
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Colorectal cancer (CRC) is the third most prevalent cancer and the second most common cause of cancer death; however, its early detection can improve the survival. Colonic polyps are considered one of the CRC's major risk factors. Throughout many biological processes and malignancies, the non-coding RNAs have essential functions. Certain long noncoding RNAs (lncRNAs), including H19, were supposed to be CRC possible biomarkers. Also, H19 has been reported to play a role in regulating the activity of beta-catenin, a protein that regulates cell-to-cell adhesion, as well as gene transcription. The current work aimed to investigate the potential significance of LncRNA H19 relative serum expression level by quantitative polymerase chain reaction (q-PCR) and beta-catenin by enzyme-linked immunosorbent assay (ELISA) as noninvasive biomarkers to discriminate between colorectal cancer and colonic polyps. The statistical analysis of the studied factors revealed that the serum expression of H19 and beta-catenin in cancer cases were substantially greater than colonic polyp cases and normal control. Conclusion: The relative expressions of H19 and beta-catenin in the serum can significantly discriminate patients with CRC from those with polyp and normal controls, which could help when screening for CRC. (AU)
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Humans , Male , Female , Adult , Middle Aged , Aged , Biomarkers, Tumor , beta Catenin , RNA, Long Noncoding , Colorectal Neoplasms/diagnosis , Early Detection of CancerABSTRACT
Background Diabetes is a major threat to public health across the world. Studies have shown that exposure to p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) is closely related to the occurrence of type 2 diabetes mellitus. However, the relevant molecular mechanism is not clear. Objective To investigate the effects of p,p'-DDE on H19 differentially methylated region (DMR) methylation and insulin secretion of rat insulinoma cells (INS-1 cells). Methods INS-1 cells were cultured with different concentrations (0, 3.125, 6.25, 12.5, 25, 50, and 75 µmol·L−1) of p,p'-DDE for 24 h, and the viability of INS-1 cells was detected by CCK-8 method. INS-1 cells were exposed to 0, 12.5, 25, and 50 µmol·L−1 p,p'-DDE for 24 h in subsequent experiments. The methylation levels of 24 CpG sites in H19 DMR were analyzed by bisulfite genomic sequencing. The expression levels of insulin-like growth factor 2 (IGF2) mRNA were detected by real-time quantitative PCR. The expression levels of IGF2 and insulin-like growth factor-1 receptor (IGF1R) proteins were detected by Western blotting. The insulin secretion function of INS-1 cells was determined by glucose-stimulatedinsulin secretion test (5 and 25 mmol·L−1 glucose, respectively). Results Compared with the control group, the viability of INS-1 cells increased significantly after treatment with 12.5 µmol·L−1 p,p'-DDE; however, it was significantly inhibited after treatment with 50 or 75 µmol·L−1 p,p'-DDE (P<0.01); therefore, 50 µmol·L−1 was chosen as the maximum concentration of exposure for subsequent experiments. The 25 µmol·L−1 p,p'-DDE treatment decreased the methylation levels of CpG18 and CpG22-CpG24 sites in H19 DMR, and the 50 µmol·L−1 p,p'-DDE treatment decreased the methylation levels of CpG10-CpG24 sites (P<0.05 or P<0.05). Multiple concentrations (12.5, 25, and 50 µmol·L−1) of p,p'-DDE down-regulated the mRNA and protein relative expression levels of IGF2 and the protein relative expression levels of IGF1R. The transcription level of IGF2 decreased to 67.8%, 68.6%, and 62.5% of the control group, the protein level of IGF2 decreased to 73.3%, 79.5%, and 80.9% of the control group, and the protein level of IGF1R decreased to 54.8%, 25.6%, and 12.9% of the control group, respectively (P<0.01). In the high glucose context, p,p'-DDE at selected concentrations inhibited the insulin secretion levels to 85.0%, 58.6%, and 49.5% of the control group, respectively (P<0.01). Conclusion p,p'-DDE could down-regulate methylation level of H19 DMR, interfere the IGF2/IGF1R signaling pathway, and inhibit insulin secretion of islet cells.
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Objective @#To investigate the role of hepatic long-chain non-coding RNA (lncRNA) H19 in key genes associated with glucose metabolism disorder induced by p,p′-dichlorodiphenyldichloroethylene (p,p′-DDE).@*Methods@#Human embryonic liver CCC-HEL-1 cells were divided into the DMSO group, 0.1 μmol/L p,p′-DDE group, 1 μmol/L p,p′-DDE group, 10 μmol/L p,p′-DDE group, small interference RNA (siRNA)+DMSO group and siRNA+10 μmol/L p,p′-DDE group. The promoter region methylation, mRNA expression and protein expression of hepatocyte nuclear factor 4α (HNF4α), forkhead box transcription factor O1 (FoxO1) and insulin-like growth factor (IGF2) were detected in CCC-HEL-1 cells using the bisulfite method, real-time fluorescence quantitative PCR (qPCR) assay and BCA assay, respectively. The changes in H19 mRNA expression, the methylation of associated genes in the promoter region and transcriptional expression were compared in CCC-HEL-1 among groups.@*Results@#Exposure to p,p′-DDE alone at different doses resulted in an increase in H19 expression, and the H19 mRNA expression was higher in the 10 μmol/L p,p′-DDE group than in the DMSO group [(1.31±0.25) vs. (1.02±0.22); P<0.05]. Lower methylation of the HNF4α gene in the pro moter region [(38.59±32.77)% vs. (61.43±24.64)%; P<0.05] and higher HNF4α mRNA expression [(1.33±0.26) vs. (1.03±0.28); P<0.05] were detected in the 10 μmol/L p,p′-DDE group than in the DMSO group, while no significant differences were detected between the two groups in terms of the methylation of FoxO1 and IGF2 genes in the promoter region, FoxO1 and IGF2 mRNA and protein expression (P>0.05). Following siRNA-induced H19 knockdown, higher methylation of the HNF4α gene in the promoter region [(71.33±22.23)% vs. (38.59±32.78)%; P<0.05], lower HNF4α mRNA expression [(0.71±0.17) vs. (1.33±0.26); P<0.05], higher methylation of FoxO1 gene in the promoter region [(47.73±34.24)% vs. (25.09±25.35)%; P<0.05] and higher IGF2 mRNA [(1.39±0.25) vs. (0.80±0.20); P<0.05] were found in the siRNA+DMSO group than in the 10 μmol/L p,p′-DDE group, and higher IGF2 protein expression was detected in the siRNA+DMSO group than in the DMSO group [(1.03±0.11) vs. (0.74±0.12); P<0.05]. @*Conclusion@#Hepatic H19 may alter HNF4α, FoxO1 and IGF2 transcription and expression through mediating the methylation of genes in the promoter region, thereby playing a role in p,p′-DDE-induced glucose metabolism disorders.
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With the increasing incidence of hematologic malignancies, exploring its pathogenesis and new treatment strategies has become new research directions. The development of high-throughput sequencing technology has led to the recognition of long non-coding RNA, which plays an important role in the development of life and the regulation of diseases. As the first reported long non-coding RNA, H19 acts as an oncogene to take part in various pathological processes in the growth and metastasis of tumors. However, the multiple roles of H19 in hematologic malignancies are lack of systematic summary. This article reviews the role of H19 in carcinogenesis, progression and drug resistance mechanism of hematologic malignancies, in order to provide guidance for further research.
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Objective:To investigate the role of lncRNA H19 in evaluating prognosis and regulating radioresistance of colon cancer, aiming to provide a new potential target for the diagnosis and treatment of colon cancer.Methods:The value of lncRNA H19 in the clinicopathological parameters and prognosis of colon cancer was assessed based on bioinformatics technology. The expression of lncRNA H19 in HCT116 and SW620 cells was regulated through siRNA and overexpression plasmid transfection, respectively. The effect of regulating lncRNA H19 expression on the proliferation, DNA synthesis, radiosensitivity and cell cycle of colon cancer cells after X-ray irradiation were detected by CCK8, EdU, cell clonogenic survival assay and flow cytometry.Results:The expression of lncRNA H19 was significantly upregulated in colon cancer tissues and correlated with poor prognosis in colon cancer patients. LncRNA H19, as a high-risk gene for colon cancer, had a significant advantage for prognostic assessment of colon cancer (AUC=0.816). Furthermore, the expression of lncRNA H19 was upregulated after X-ray irradiation in colon cancer cells. Knockdown of lncRNA H19 (siRNA-H19) significantly increased the radiosensitivity in HCT116 cells, while overexpression of lncRNA H19 (H19-OE) enhanced the radioresistance in SW620 cells. Moreover, flow cytometry revealed that the G 2/M phase arrest induced by X-ray irradiation was obviously aggravated after siRNA-H19 treatment in colon cancer cells, which suggested that lncRNA H19 might regulate the radiosensitivity by inhibiting cell cycle progression. Conclusion:LncRNA H19 plays a key role in the prognostic assessment and regulating the radiosensitivity in colon cancer, which can be used as a potential target for improving radiosensitivity of colon cancer radiotherapy.
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Objective To investigate the expression and the potential roles of long non-coding RNA(lncRNA)cancer susceptibility candidate 2(CASC2)and imprinted gene H19 in extrahepatic cholangiocarcinoma(ECC). Methods Four samples from patients with ECC were collected for high-throughput sequencing which was conducted to reveal the transcriptomic profiles of lncRNA CASC2 and H19.Bioinformatics tools were employed to predict the potential roles of the two genes.Another 22 ECC tissue samples and the cholangiocarcinoma cell lines(RBE,QBC939,HuH-28,and HuCCT1)with different degrees of differentiation were selected for validation.The para-carcinoma tissue and normal human intrahepatic biliary epithelial cell(HIBEC)were used as the control groups.The expression levels of lncRNA CASC2 and H19 in carcinoma tissue,para-carcinoma tissue,and cell lines were determined by real-time quantitative polymerase chain reaction(qRT-PCR).The correlation analysis was carried out for the clinical indicators of patients with the expression levels of the target genes. Results The two target genes showed significantly different expression between carcinoma tissue and para-carcinoma tissue(all P<0.05).Specifically,CASC2 had higher expression level in the carcinoma tissue than in the para-carcinoma tissue(t=1.262,P=0.025),whereas the expression of H19 showed an opposite trend(t=1.285,P=0.005).The expression levels of CASC2 in QBC939(t=8.114,P=0.015)and HuH-28(t=9.202,P=0.012)cells were significantly higher than that in the control group.The expression levels of H19 were significantly lower in RBE(t=-10.244,P<0.001),QBC939(t=-10.476,P<0.001),HuH-28(t=-19.798,P<0.001),and HuCCT1(t=-16.193,P=0.004)cells than in the control group.Bioinformatics analysis showed that CASC2 was mainly involved in the metabolic process and H19 in the development of multicellular organisms.Both CASC2 and H19 were related to catalytic activity.The expression level of lncRNA CASC2 was correlated with pathological differentiation(χ 2=6.222,P=0.022)and lymph node metastasis(χ2=5.455,P=0.020),and that of lncRNA H19 with pathological differentiation(χ2=1.174,P=0.029)and tumor size(χ2=-0.507,P=0.037). Conclusions In the case of ECC,lncRNA CASC2 and H19 have transcription disorders.lncRNA CASC2 is generally up-regulated in the carcinoma tissue,while H19 is down-regulated.Both genes have the potential to become new molecular markers for ECC.
Subject(s)
Humans , Bile Duct Neoplasms/genetics , Bile Ducts, Intrahepatic/metabolism , Cholangiocarcinoma/genetics , Gene Expression Regulation, Neoplastic , RNA, Long Noncoding/genetics , Tumor Suppressor Proteins/geneticsABSTRACT
Abstract Objective Tongue squamous cell carcinoma (TSCC) is an oral cancer, with high malignancy and frequent early migration and invasion. Only a few drugs can treat tongue cancer. Ginsenoside Rd is a ginseng extract with anti-cancer effects. Many noncoding RNAs are abnormally expressed in tongue cancer, thus influencing its occurrence and development. H19 and miR-675-5p can promote cancer cell growth. This study aimed to analyze the regulation effect of ginsenoside Rd on H19 and miR-675-5p in tongue cancer. Methodology We used CCK8 and flow cytometry to study the growth and apoptosis. Transwell assay was used to assess invasion; wound-healing assay to assess migration; and colony formation assays to test the ability of cells to form colonies. H19, miR-675-5p, and CDH1 expressions were analyzed by qPCR. E-cadherin expression was detected using western blot. CRISPR/cas9 system was used for CDH1 knockout. Results Ginsenoside Rd inhibited the growth and increased the apoptosis of SCC9 cells. Ginsenoside Rd also inhibited the migration and invasion of SCC9 cells. H19 and miR-675-5p were highly expressed, while CDH1 and E-cadherin expressions were low. H19 and miR-675-5p promoted SCC9 metastasis. In contrast, CDH1 and E-cadherin inhibited the metastasis of SCC9 cells. Bioinformatics analysis showed that miR-675-5p was associated with CDH1. H19 and miR-675-5p expressions decreased after ginsenoside Rd treatment, while CDH1 and E-cadherin expressions increased. Conclusions Ginsenoside Rd inhibits tongue cancer cell migration and invasion via the H19/miR-675-5p/CDH1 axis.
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Objective:To investigate the expression and correlation of H19, matrix metalloproteinase (MMP)-2 and MMP-9 in patients with recurrent spontaneous abortion (RSA).Methods:Human extravillous trophoblast cell line HTR-8 was cultured in vitro. Lentivirus was used to infect the HTR-8 cell line to over-express or knockdown the expression of H19. The concentrations of MMP-2 and MMP-9 protein in cell culture supernatant were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Spearman correlation analysis was used to understand the correlation between H19 and the expression levels of MMP-2 and MMP-9. Results:After overexpression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-ph19 group were significantly higher than those in Lv-vector group ( P<0.05); After interfering with the expression of H19, the expression levels of MMP-2 and MMP-9 mRNA and protein concentration in Lv-shH19 group were significantly lower than those in Lv-shcon control group ( P<0.05). The number of spontaneous abortions in patients with recurrent spontaneous abortion was significantly higher than that in the control group ( P<0.05). qRT-PCR showed that the expression levels of H19, MMP-2 and MMP-9 mRNA in villi of patients with RSA were significantly lower than those in the control group ( P<0.05). There was a positive correlation between H19 and the expression levels of MMP-2 and MMP-9 ( P<0.05). Conclusions:H19 regulates the expression of MMP-2 and MMP-9 of trophoblast during early pregnancy, and the abnormal expression of H19, MMP-2, and MMP-9 in human first-trimester villous tissues was related with the incidence of early miscarriage.
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Objective:To investigate the molecular mechanism of antiglioma effect of curcumin.Methods:Cell experiment: (1) U251MG and SHG-44 cells at logarithmic growth phase were treated with 10 μmol/L curcumin (curcumin group) or same volume of dimethyl sulfoxide solution (control group); cells were transfected with negative control small interfering RNA (siRNA) and long non-coding RNA (lncRNA) H19 siRNA (negative control siRNA group and H19 siRNA group); cells were transfected with negative control siRNA and H19 siRNA, respectively, and then, they were treated with 10 μmol/L curcumin (negative control siRNA+curcumin group and H19 siRNA+curcumin group); the H19 siRNA was co-transfected with negative control miR inhibitor or miR-491-5p inhibitor into these cells (H19 siRNA+negative control inhibitor group and H19 siRNA+miR-4915p inhibitor group); H19 siRNA+negative control miR inhibitor or H19 siRNA+miR-491-5p inhibitor were co-transfected into the cells, and then, they were treated with 10 μmol/L curcumin (H19 siRNA+negative control inhibitor+curcumin group and H19 siRNA+miR-491-5p inhibitor+curcumin group); the cells were co-transfected with miR-491-5p mimic+blank plasmid or miR-491-5p mimic+HOXA9 overexpression plasmid, and then they were treated with 10 μmol/L curcumin (miR-491-5p mimic+blank plasmid+curcumin group and miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group); real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of H19, miR-491-5p, and HOXA9; CCK-8 assay was used to detect the cell proliferation; flow cytometry was used to detect the cell apoptosis; plate cloning method was employed to detect the number of cell clone formation; Transwell assay was used to detect the cell migration; and the HOXA9 protein expression was measured by Western blotting. (2) The 293T cells at the logarithmic growth phase were chosen; the negative control miRNA mimics or miR-491-5p mimics combined with wild-type H19, wild-type HOXA9 3'-UTR plasmid vectors were co-transfected into the cells, respectively (negative control mimic+wild type H19 group and miR-491-5p mimic+wild type H19 group, negative control mimic+wild type HOXA9 3'-UTR group and miR-491-5p mimic+wild type HOXA9 3'-UTR group); the luciferase activity was detected by dual luciferase reporter experiment. (3) Thirty specimens from glioma patients (glioma group) underwent surgical resection and pathologically confirmed in our hospital from May 2017 to May 2019 and 30 normal brain tissue specimens obtained during decompression (normal group) at the same period were chosen; the mRNA expressions of H19, miR-491-5p, and HOXA9 were detected by qRT-PCR, and the HOXA9 protein expression level in these specimens was detected by Western blotting. (4) Twenty-four nude mice were randomly divided into negative control short hairpin RNA (shRNA) group, H19 shRNA group, negative control shRNA+curcumin group, and H19 shRNA+curcumin group ( n=6); U251MG cells stably transfected with negative control shRNA or H19 shRNA were intraperitoneally injected, respectively, into the mice; and 60 mg/kg curcumin was injected on the next d; the tumor volume was measured on the 7 th, 11 th, 15 th, 19 th, 23 rd, and 27 th d of rearing; and the H19, miR-491-5p and HOXA9 mRNA expressions in the tumor tissues were detected by qRT-PCR; the HOXA9 protein expression was detected by Western blotting. Results:(1) When curcumin group comparing with control group, and H19 siRNA group comparing with negative control siRNA group, U251MG and SHG-44 cells had significantly decreased miR-491-5p mRNA and protein expressions, and significantly increased miR-491-5p mRNA expression ( P<0.05); as compared with that in the H19 siRNA+negative control inhibitor group, the HOXA9 mRNA and protein expressions in U251MG and SHG-44 cells of H19 siRNA+miR-491-5p inhibitor group were significantly higher ( P<0.05). When curcumin group comparing with control group, H19 siRNA group comparing with negative control siRNA group, H19 siRNA+curcumin group comparing with negative control siRNA+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly decreased cell proliferation rate, significantly increased apoptosis rate, significantly reduced number of cell clone formation, and significantly reduced cell migration number ( P<0.05). When H19 siRNA+miR-491-5p inhibitor+curcumin group comparing with H19 siRNA+negative control inhibitor+curcumin group, miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group comparing with miR-491-5p mimic+blank plasmid+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly increased cell proliferation rate, significantly reduced apoptosis rate, significantly increased number of cell clone formation, and significantly increased cell migration number ( P<0.05). (2) When miR-491-5p mimic+wild-type H19 group comparing with negative control mimic+wild-type H19 group, miR-491-5p mimic+wild-type HOXA9 3'-UTR group comparing with negative control mimic+wild-type HOXA9 3'-UTR group, the cell luciferase activity was significantly reduced ( P<0.05). (3) As compared with those in the normal group, the H19 and HOXA9 mRNA expressions and HOXA9 protein expression in the glioma group were significantly increased, and the miR-491-5p mRNA expression was significantly reduced ( P<0.05). (4) On the 27 th d of rearing, when H19 shRNA group comparing with negative control shRNA group, and H19 shRNA+curcumin group comparing with negative control shRNA+curcumin group, the tumor volume was significantly reduced, the miR-491-5p mRNA expression in the tumor tissues was significantly increased, and the H19 mRNA, HOXA9 mRNA and protein expressions were significantly reduced ( P<0.05). Conclusion:Curcumin may inhibit the cell proliferation and migration and promote the apoptosis of glioma cells through lncRNA H19/miR-491-5p/HOXA9 axis.
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Long noncoding RNA (lncRNA) are defined as noncoding RNA with a length of more than 200 nucleotides, which can regulate a variety of cellular processes. LncRNA H19 is widely involved in the pathophysiological process of intestinal inflammation and intestinal cancer by regulating the function of intestinal mucosal barrier. This article reviewed the effect of lncRNA H19 on intestinal mucosal barrier under different intestinal states and inflammation-cancer related signaling pathway.
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OBJECTIVES: To investigate the predictive value of long non-coding RNA (lncRNA) H19 and the ten-eleven translocation enzyme 1 (TET1) transcriptional expression in postoperative recurrence of uterine fibroids (UFs). METHODS: Seventy-five patients with UF, who underwent surgical treatment, were enrolled in the treatment group, and 60 healthy individuals were enrolled in the control group. The relative expression levels of lncRNA H19 and TET1 mRNA in the serum and UF tissues were analyzed. The patients were further divided into a better curative (BC) group and a poor efficacy (PE) group to analyze the predictive value of lncRNA H19 and TET1 and the independent risk factors affecting the recurrence of UF. RESULTS: Compared with the control group, lncRNA H19 expression levels were significantly higher, while TET1 expression levels were significantly lower in the treatment group (p<0.001). The area under the receiver operating characteristic (ROC) curve (AUC) values of the two indicators for diagnostic importance were found to be 0.872 and 0.826, respectively. Compared with the PE group, lncRNA H19 expression levels were significantly lower, while TET1 expression levels were significantly higher in the BC group (p<0.001). The AUC values of the two indicators for their predictive efficacy were 0.788 and 0.812, respectively. Logistic regression analysis showed that age, menarche age, maximum diameter of UFs, number of UFs, lncRNA H19 levels, and TET1 levels were independent risk factors affecting UF recurrence. The AUC values of lncRNA H19 and TET1 for their predictive value for postoperative recurrence were 0.814 and 0.765, respectively. CONCLUSIONS: The lncRNA H19 and TET1 have high diagnostic and predictive efficacy for determining the postoperative recurrence of UFs.
Subject(s)
Humans , Female , RNA, Long Noncoding/genetics , Leiomyoma , RNA, Messenger , ROC Curve , Proto-Oncogene Proteins , Mixed Function Oxygenases , Neoplasm Recurrence, LocalABSTRACT
The long noncoding RNA (lncRNA) H19 is involved in the pathogenesis of endometriosis by modulating the proliferation and invasion of ectopic endometrial cells in vitro, but related in vivo studies are rare. This study aimed to investigate the role of lncRNA H19 in a nude mouse model of endometriosis. Ectopic endometrial stromal cells (ecESCs) were isolated from ectopic endometrium of patients with endometriosis and infected with lentiviruses expressing short hairpin RNA (shRNA) negative control (LV-NC-shRNA) or lncRNA-H19 shRNA (LV-H19-shRNA). The ecESCs infected with LV-NC-shRNA and LV-H19-shRNA were subcutaneously implanted into forty 6- to 8-week-old female nude mice. The size and weight of the endometriotic implants were measured at 1, 2, 3, and 4 weeks after implantation and compared, and lncRNA H19 levels in endometriotic implants were evaluated using real-time polymerase chain reaction (RT-PCR). All nude mice survived the experimental period, and no significant differences in body weight were observed between the experimental group and the control group. All nude mice developed histologically confirmed subcutaneous endometriotic lesions with glandular structures and stroma after 1 week of implantation. The subcutaneous lesions in the LV-NC-shRNA group after 1, 2, 3, and 4 weeks of implantation were larger than those in the LV-H19-shRNA group, and lncRNA H19 levels in subcutaneous lesions in the LV-NC-shRNA group were significantly higher than those in the LV-H19-shRNA group. Knockdown of lncRNA H19 suppresses endometriosis in vivo. Further study is required to explore the underlying mechanism in the future.
Subject(s)
Humans , Animals , Female , Rabbits , Endometriosis/genetics , RNA, Long Noncoding/genetics , RNA, Small Interfering/genetics , Cell Proliferation/genetics , Endometrium , Mice, NudeABSTRACT
Somatic Cell Nuclear Transfer (SCNT-Cloning) is a promising technique in many areas and is based on genetically identical individuals. However, its efficiency is low. Studies suggest that the leading cause is inadequate epigenetic reprogramming. This study aimed to characterize the methylation pattern of the exon 10 regions of the IGF2 gene and the Imprinting Control Region (ICR) of the H19 gene in the placenta of cloned calves. For this study, female and male cloned calves presenting different phenotypes were used. Genomic DNA from these animals' placenta was isolated, then treated with sodium bisulfite and amplified to the ICR/H19 and IGF2 loci. PCR products were cloned into competent bacteria and finally sequenced. A significant difference was found between controls and clones with healthy phenotypes for the ICR/H19 region. In this region, controls showed a hemimethylated pattern, as predicted in the literature due to this region has an imprinted control, while clones were showed less methylated. For the IGF2, no significant differences were found between controls and clones. These results suggest that different genomic regions in the genome may be independently reprogrammed and that failures in reprogramming the DNA methylation patterns of imprinted genes may be one of the causes of the low efficiency of SCNT.(AU)
A Transferência Nuclear de Células Somáticas (TNCS-Clonagem) é uma técnica promissora em várias áreas, e se baseia na produção de indivíduos geneticamente idênticos. No entanto, sua eficiência é baixa. Estudos sugerem que a principal causa seja uma reprogramação epigenética inadequada. O objetivo desse trabalho é caracterizar o padrão de metilação da região éxon 10 do gene IGF2 e da Região Controladora de Imprinting (ICR) do gene H19 na placenta de bezerros clonados. Para a execução do trabalho foram selecionados clones bovinos fêmeas e machos, apresentando diferentes fenótipos. O DNA da placenta desses animais foi extraído, e em seguida foi tratado com bissulfito de sódio e amplificado para os loci ICR/H19 e IGF2. Os produtos da PCR foram clonados em bactérias competentes e, por fim, sequenciados. Foi encontrada uma diferença significativa entre os controles e os clones com fenótipos saudáveis para a região da ICR/H19. Nesta região, os controles tiveram um padrão hemimetilado, como previsto pela literatura, devido essa região ser imprinted. Enquanto os clones encontravam-se menos metilados. Para a região do éxon 10 do IGF2, não foi encontrada diferença significativa entre controles e clones. Estes resultados sugerem que as diferentes regiões do genoma podem se reprogramar independente umas das outras e que falhas na reprogramação do padrão de metilação do DNA de genes imprinted podem ser uma das causas da baixa eficiência da TNCS.(AU)
Subject(s)
Animals , Cattle , Placenta , Cattle/genetics , Clone Cells , Epigenomics , Insulin-Like Growth Factor II/analysis , DNA MethylationABSTRACT
The osteogenic differentiation of mesenchymal stem cells (MSCs) has potential clinical values in the treatmentof bone-related diseases. Long non-coding RNA H19 and microRNA-140-5p (miR-140-5p) have attractedmuch attention of researchers by virtue of their biological importance in cell differentiation and bone formation. Moreover, bioinformatics analyses suggest that miR-140-5p have the potential to bind with H19 andSATB homeobox 2 (SATB2). In this study, we further explored whether H19 could regulate osteogenicdifferentiation of human bone marrow-derived MSCs (BM-MSCs) by miR-140-5p/SATB2 axis. RT-qPCRassay was conducted to examine the expression of H19, miR-140-5p and SATB2. The osteogenic differentiation capacity of BM-MSCs was assessed through alkaline phosphatase (ALP) activity and osteogenic markerexpression. The relationships among H19, miR-140-5p and SATB2 were examined through bioinformaticsanalyses, luciferase reporter assay, RIP assay and RNA pull-down assay. H19 expression was remarkablyincreased and miR-140-5p expression was dramatically reduced during osteogenic differentiation of BMMSCs. Functional analyses revealed that H19 overexpression or miR-140-5p depletion accelerated osteogenicdifferentiation of BM-MSCs. Conversely, H19 loss or miR-140-5p increase suppressed osteogenic differentiation of BM-MSCs. MiR-140-5p was confirmed as a target of H19, and miR-140-5p could bind to SATB2 aswell. Moreover, H19 knockdown reduced SATB2 expression by upregulating miR-140-5p. Additionally, miR140-5p depletion antagonized the inhibitory effect of H19 knockdown on osteogenic differentiation of BMMSCs. And, miR-140-5p inhibited osteogenic differentiation of BM-MSCs by targeting SATB2. In conclusion,H19 promoted osteogenic differentiation of BM-MSCs through regulating miR-140-5p/SATB2 axis, deepeningour understanding on the molecular mechanisms of H19 in coordinating osteogenesis.
ABSTRACT
Abstract Shiga toxin-producing Escherichia coli (STEC) are a heterogeneous group of foodborne pathogens causing a broad spectrum of human disease, from uncomplicated diarrhea to hemolytic uremic syndrome (HUS). In this study, we report an HUS case associated with an O59:NM H19 mstrain, harboring stx2a, iha, lpfAO26, lpfAO113 genes associated with STEC, and aatA, aap, pic, sigA, agg4A genes associated with enteroaggregative E. coli (EAEC), named Stx-EAEC. The strain showed low toxicity on Vero cells, and was resistant to streptomycin and trimethoprim/sulfonamides. The child carried the bacteria for more than 100 days. Since the large outbreak associated with Stx-EAEC O104:H4, many strains with similar profiles have been described. In Germany, an O59:NM[H19] strain, with comparable characteristics to the Argentine strain, was isolated from a bloody diarrhea case. In Argentina, this is the first report of an HUS case associated with a Stx-EAEC infection, and represents a new challenge for the surveillance system. © 2019 Published by Elsevier Espana, S.L.U. on behalf of Asociacion Argentina de Microbiolog´a. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/ licenses/by-nc-nd/4.0/).
Resumen Escherichia coli productor de la toxina Shiga (STEC) es un grupo heterogéneo de patógenos transmitidos por alimentos que causan un amplio espectro de enfermedades humanas, desde diarrea no complicada hasta síndrome urémico hemolítico (SUH). Nosotros informamos de un caso de SUH por O59:NM[H19], que portaba los genes stx2a, iha, lpfAo26, lpfAoii3 asociados con STEC, y los genes aatA, aap, pic, sigA, agg4A de E. coli enteroagregativo (EAEC), llamado EAEC-Stx. La cepa mostró baja citotoxicidad en las células Vero, y fue resistente a estreptomicina y trimetoprima/sulfonamidas. El niño excretó la bacteria durante más de 100 días. Desde el brote asociado con EAEC-Stx O104:H4, se describieron muchas cepas con perfiles similares. En Alemania se aisló una cepa O59:NM[H19] de una diarrea sanguinolenta, con características comparables a la cepa argentina. Este es el primer informe de un caso de SUH asociado a una infección por EAEC-Stx, y representa un nuevo desafío para el sistema de vigilancia. © 2019 Publicado por Elsevier Espana, S.L.U. en nombre de Asociación Argentina de Microbiología. Este es un artículo Open Access bajo la licencia CC BY-NC-ND (http://creativecommons. org/licenses/by-nc-nd/4.0/).