Knockdown of circRNA WD repeat containing protein 1 inhibits proliferation and induces apoptosis of chondrocytes in knee osteoarthritis / 中国组织工程研究

BACKGROUND:Circular RNAs(circRNAs)play important roles in a variety of diseases or tumors,and recent findings have revealed that circRNAs are abnormally expressed in knee osteoarthritis and are involved in disease progression through microRNA/mRNA regulation. OBJECTIVE:To investigate the effect of circRNA WD repeat containing protein 1(circ-BRWD1)/miR-488-3p/DNA methyltransferase 3A(DNMT3A)on the proliferation and apoptosis of chondrocytes in knee osteoarthritis. METHODS:Real-time fluorescence quantitative PCR was performed to detect the expression of circ-BRWD1,miR-488-3p,DNMT3A in knee osteoarthritis chondrocytes.Cells were divided into si-NC group,si-circ-BRWD1 group,vector group,circ-BRWD1 group,si-circ-BRWD1+anti-miR-NC group,si-circ-BRWD1+anti-miR-488-3p group,miR-NC group,miR-488-3p group,anti-miR-NC group,anti-miR-488-3p group,miR-488-3p+vector group,miR 488-3p+DNMT3A group.Real-time fluorescence quantitative PCR was used to detect circ-BRWD1,miR-488-3p,DNMT3A expression,MTT and flow cytometry assay were used to detect cell proliferation and apoptosis.Western blot assay was used to detect DNMT3A and proliferation/apoptosis-related protein expression.Dual luciferase reporter assay was used to Dual luciferase reporter assay to detect the targeting relationship of circ-BRWD1 with miR-488-3p and miR-488-3p with DNMT3A. RESULTS AND CONCLUSION:circ-BRWD1 and DNMT3A were highly expressed and miR-488-3p was lowly expressed in knee osteoarthritis chondrocytes compared with normal chondrocytes.Knockdown of circ-BRWD1 or overexpression of miR-488-3p inhibited proliferation and induced apoptosis in knee osteoarthritis chondrocytes.circ-BRWD1 targeted negative regulation of miR-488-3p and inhibition of miR-488-3p reversed the effect of circ-BRWD1 knockdown on chondrocyte proliferation and apoptosis in knee osteoarthritis.miR-488-3p targeted negative regulation of DNMT3A and upregulation of DNMT3A reversed the effect of miR-488-3p overexpression on chondrocyte proliferation and apoptosis in knee osteoarthritis.circ-BRWD1 could regulate the expression of DNMT3A by regulating miR-488-3p.To conclude,knockdown of circ-BRWD1 inhibits chondrocyte proliferation and induces apoptosis in knee osteoarthritis,and the mechanism of action may be related to the regulation of miR-488-3p/DNMT3A axis.

Advance in the immunodeficiency-centromeric instability-facial anomalies syndrome / 中华实用儿科临床杂志

The immunodeficiency-centromeric instability-facial anomalies (ICF) syndrome is the only known human genetic disease involving DNA methylation defects.About 50% of the cases are caused by the compound heterozygous mutation of DNMT3B gene.About a hundred cases were reported in the world, but only a few cases came from China.There may be a misdiagnosis and missed diagnosis.To the best of our knowledge, no Chinese article systematically discusses the ICF syndrome.This paper aims to review the possible mechanisms, clinical manifestations, genetic characteristics, treatment, and prognosis of the ICF syndrome, and to improve Chinese doctors′ knowledge about this disease.

Mechanism of heroin promoting addiction and relapse in progeny of male rats by inhibiting promoter methylation of imprinted gene H19 by down-regulating DNMT3a / 中国药理学通报

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

Effects of DNMT3A on mouse cardiac fibroblast proliferation and migration under high glucose environment / 中国药理学通报

Aim To investigate the effect of DNA methyltransferase 3A (DNMT3A) on the proliferation and migration of cardiac fibroblasts (CFs) in C57 mice under high glucose environment. Methods The hearts of C57 mice were taken from 1 to 3 days. After cutting and digesting, CFs were extracted by differential adherance centrifugattion and observed under microscope. After cell attachment, the cells were cultured under low glucose (5.5 mmol • L

Overexpression of lncRNA-MEG3 inhibits endometrial cell proliferation and invasion via miR-21-5p/DNMT3B/Twist

Abstract Recent studies have found that lncRNA-MEG3(MEG3) plays an important role in the development of EMs (Endometriosis), but the specific mechanism needs to be further explored. This study aimed to investigate the effect of MEG3 on the proliferation, invasion of EMs cells. The authors used RT-qPCR to detect the expression of MEG3 and miR-21-5p in EMs tissues and hESCs cells, MTT and Transwell to detect cell proliferation and invasion, western blotting assay to detect the expression of DNMT3B and Twist, MSP to detect the methylation of Twist. The present study's detection results showed that MEG3 was lowly expressed in EMs tissues and hESCs cells, and overexpression of MEG3 could down-regulate miR-21-5p and inhibit endometrial cell proliferation and invasion. In addition, overexpression of MEG3 upregulated the expression of DNMT3B and promoted the methylation of TWIST. In conclusion, the present findings suggest that MEG3 is downregulated in EMs tissues, and overexpression of MEG3 can promote the activity of DNA methyltransferase DNMT3B by downregulating miR-21-5p, thereby promoting the methylation of Twist, downregulating Twist level to inhibits hESCs cells proliferation and invasion.

Role of DNMT3a in Hydroquinone-Induced Hematopoietic Stem Cell Toxicity / 中国实验血液学杂志

OBJECTIVE@#To investigate the regulatory effect and mechanism of DNA methyltransferase 3A (DNMT3a) in hydroquinone-induced hematopoietic stem cell toxicity.@*METHODS@#Cells (HSPC-1) were divided into 4 groups, that is A: normal HSPC-1; B: HQ-intervented HSPC-1; C: group B + pcDNA3 empty vector; D: group B + pcDNA3- DNMT3a. RT-qPCR and Western blot were used to detect the expression levels of DNMT3a and PARP-1 mRNA and protein, respectively. Cell morphology was observe; Cell viability and apoptosis rate of HSPC-1 were detected by MTT and flow cytometry, respectively.@*RESULTS@#Compared with group A, the expression levels of DNMT3a mRNA and protein in HSPC-1 of group B were decreased, while PARP-1 mRNA and protein were increased (P<0.05); there was no significant difference in the above indexes between group C and group B; compared with group B, the expression levels of DNMT3a mRNA and protein showed increased, while PARP-1 mRNA and protein were decreased significantly in cells of group D transfected with DNMT3a (P<0.05). Cells in each group were transfected with DNMT3a and cultured for 24 h, HSPC-1 in group A showed high density growth and mononuclear fusion growth, while the number of HSPC-1 in group B and C decreased and grew slowly. Compared with group B and C, the cell growth rate of group D was accelerated. The MTT analysis showed that cell viability of HSPC-1 in group B were lower than that of group A at 24 h, 48 h and 72 h (P<0.05); after transfected with DNMT3a, the cell viability of HSPC-1 in group D were higher than that of group B at 24 h, 48 h and 72 h (P<0.05). The apoptosis rate of cells in group B was significantly higher than that of group A (P<0.001), while the apoptosis rate in group D was lower than that of group B (P<0.001).@*CONCLUSION@#DNMT3a may be involved in the damage of hematopoietic stem cells induced by hydroquinone, which may be related to the regulation of PARP-1 activity by hydroquinone-inhibited DNMT3a.

Aging-elevated inflammation promotes DNMT3A R878H-driven clonal hematopoiesis

Aging-elevated DNMT3A R882H-driven clonal hematopoiesis (CH) is a risk factor for myeloid malignancies remission and overall survival. Although some studies were conducted to investigate this phenomenon, the exact mechanism is still under debate. In this study, we observed that DNMT3A R878H bone marrow cells (human allele: DNMT3A R882H) displayed enhanced reconstitution capacity in aged bone marrow milieu and upon inflammatory insult. DNMT3A R878H protects hematopoietic stem and progenitor cells from the damage induced by chronic inflammation, especially TNFα insults. Mechanistically, we identified that RIPK1-RIPK3-MLKL-mediated necroptosis signaling was compromised in R878H cells in response to proliferation stress and TNFα insults. Briefly, we elucidated the molecular mechanism driving DNMT3A R878H-based clonal hematopoiesis, which raises clinical value for treating DNMT3A R882H-driven clonal hematopoiesis and myeloid malignancies with aging.

Effects of DNMT3A regulating Drp1 on proliferation and migration of activated hepatic stellate cells / 中国药理学通报

Aim To investigate the effects of DNMT3A regulating Drp1 mediated mitochondrial fission on the proliferation and migration of active hepatic stellate cells. Methods HSC-T6 cells were activated by 5 μg·L-1 TGF-β1 for 24 h, and DNMT3A lentivirus infection model was established to silence DNMT3A. The experiment was divided into control group, TGF-β1 group, TGF-β1+LV5-NC group and TGF-β1+ LV5-DNMT3A group. The effects of DNMT3A on related mRNA and protein expression were detected by RT-qPCR and Western blot. The cell proliferation was detected by CCK-8. The effect of DNMT3A on the migration ability of HSCs cells was observed by Wound healing assay and Transwell migration assay. Results Lentivirus infection successfully constructed a DNMT3A silencing model. Compared with the control group, the level of DNMT3A significantly increased, the mRNA and protein levels of the fibrosis markers collagen and α-SMA in the TGF-β1 group significantly increased, and the mRNA and protein levels of the mitochondrial fission marker Drp1 significantly increased; At the same time, the proliferation and migration ability of HSCs cells was significantly improved. Compared with the NC group, the DNMT3A level of the DNMT3A silenced group was significantly reduced, the expressions of collagen I, α-SMA and Drp1 were significantly inhibited, and the proliferation and migration capabilities of HSCs were also significantly inhibited. Conclusions Silencing DNMT3A inhibits the level of Drp1 and inhibits the proliferation and migration of HSCs at the same time. It is suggested suggest that DNMT3A-mediated low level DNA methylation modification may inhibit the occurrence of mitochondrial fission by inhibiting the level of Drp1, thereby inhibiting the activation of HSCs and affecting the occurrence and development of liver fibrosis. ,,,,.

Expression of DNMT3b in human bladder cancer tissue and its correlation with clinical prognosis / 南方医科大学学报

OBJECTIVE@#To investigate the expression of DNMT3b in human bladder cancer tissues and its correlation with postoperative survival of patients with bladder cancer.@*METHODS@#Thirty-eight pairs of surgically resected human bladder cancer tissues and adjacent bladder tissues were detected by immunohistochemistry for DNMT3b expression, and the correlations of DNMT3b expression level were analyzed with the patients'age, gender, pathological grade, tumor size, T stage, lymph node metastasis and TNM stages. Kaplan-Meier survival analysis was performed to assess the effect of DNMT3b expression on survival outcomes of the patients.@*RESULTS@#High DNMT3b protein expression was detected in 63.16% of the bladder cancer tissues and in 13.16% of the adjacent tissues ( < 0.05). The expression level of DNMT3b was associated with the pathological grade (=0.002), tumor size ( < 0.001), T stage ( < 0.001), lymphatic metastasis (=0.039) and TNM stage ( < 0.001), but not with gender or age of the patients. Multivariate logistic regression analysis showed that the protein expression level of DNMT3b was correlated with tumor size (=0.008) and TNM grades of the tumor (=0.042). Kaplan-Meier analysis showed that the patients with a high DNMT3b expression had a significantly shorter overall survival than those with a low DNMT3b expression (=0.021).@*CONCLUSIONS@#DNMT3b overexpression in bladder cancer is closely related to such clinicopathological factors as pathological grade, tumor size, T stage, lymphatic metastasis, and TNM stage and a shorter overall survival of the patients, suggesting the potential value of DNMT3b as a prognostic marker and a new therapeutic target for bladder cancer.

First identified Korean family with Tatton-Brown-Rahman Syndrome caused by the novel DNMT3A variant c.118G>C p.(Glu40Gln)

Tatton-Brown-Rahman Syndrome (TBRS), an overgrowth syndrome caused by heterozygous mutation of DNMT3A, first was described in 2014. Approximately 60 DNMT3A variants, including 32 missense variants, have been reported, with most missense mutations located on the DNMT3A functional domains. Autosomal dominant inheritance by germ-line mutation of DNMT3A has been reported, but vertical transmission within a family is extremely rare. Herein, we report the first Korean family with maternally inherited TBRS due to the novel heterozygous DNMT3A variant c.118G>C p.(Glu40Gln), located outside the main functional domain and identified by multigene panel sequencing. The patient and her mother had typical clinical features, including tall stature during childhood, macrocephaly, intellectual disability, and characteristic facial appearance. TBRS shows milder dysmorphic features than other overgrowth syndromes, potentially leading to underdiagnosis and underestimated prevalence; thus, targeted multigene panel sequencing including DNMT3A will be a useful tool in cases of overgrowth and unexplained mild intellectual disability for early diagnosis and genetic counseling.

High expression of DNMT3B promotes proliferation and invasion of hepatocellular carcinoma cells Hippo signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the role of DNMT3B in regulating the proliferation and invasion of hepatocellular carcinoma (HCC) cells.@*METHODS@#We collected the tumor tissues and adjacent tissues from a total of 175 patients with HCC diagnosed in the Second Affiliated Hospital of Chongqing Medical University between May, 2008 and May, 2013 to prepare the tissue microarrays. The association of the expression of DNMT3B with the prognosis and the tumor-free survival and tumor-specific survival rates of the patients was analyzed. Univariate and multivariate Cox regression analyses were used to analyze the effect of DNMT3B expression on the prognosis of HCC. We used RNA interference technique to knock down the expression of DNMT3B in Huh-7 hepatoma cells and observed the changes in cell proliferation using CCK-8 assay and EDU staining and in cell migration and invasion ability using Transwell assay.@*RESULTS@#The positive rates of DNMT3B was significantly higher in HCC tissues than in paired adjacent tissues (67.4% 41.1%, =0.015). A high DNMT3B expression in HCC was significantly associated with the tumor size (=0.001), vascular invasion (=0.004), and intrahepatic metastasis (=0.018). The patients with high DNMT3B expressions had significantly lower tumor-free and tumor-specific survival rates than those with low DNMT3B expressions ( < 0.005). In Huh-7 cells, silencing DNMT3B significantly inhibited the cell proliferation and inhibited cell migration and invasion. Western blotting showed that silencing DNMT3B obviously increased LATS1 expression, decreased the expression of YAP1, and activated Hippo signaling pathway. Methylation-specific PCR showed that the methylation level of LATS1 was decreased in the cells with DNMT3B silencing.@*CONCLUSIONS@#The expression level of DNMT3B is significantly higher HCC tissues than in the adjacent tissues, and the high expression of DNMT3B is closely related to the low survival rate of the patients. Silencing DNMT3B inhibits the proliferation, migration and invasion of HCC cells. DNMT3B promotes the progression of HCC primarily by enhancing the expression of YAP1 through methylation of LATS1 and inhibition of its expression, which inhibits the anti-cancer effect of Hippo signaling pathway.

Impact of DNMT3A Gene Mutation on Response of Acute Myeloid Leukemia Patients to Induction Therapy

Aim: To evaluate the frequency and prognostic impact of DNA methyltransferase 3A (DNMT3A) gene mutation on response to induction therapy in newly diagnosed acute myeloid leukemia patients. Study Design: Cross-sectional descriptive study. Place and Duration: Hematology units of Suez Canal and Ain Shams schools of Medicine, Egypt. Between September 2016 and July 2017. Methodology: The study enrolled forty patients (male: female ratio was 1; mean age was 52.4 ± 19.4 years) with newly diagnosed de novo AML before starting induction therapy. DNMT3A mutations were detected using dye terminator sequencing technique for the second part of DNMT3A, encompassing the PHD and methyltransferase domains and representing exons 11 till the last exon 23. Hematological, cytogenetic studies and DNMT3A mutation results were compared to the patients’ hematological response to induction therapy. Results: Fourteen patients (35%) of the study participants had DNMT3A mutations while 65% had the wild type. Approximately 49.5% of mutations occurred in exon 23, the most common mutations were (R882C and R882H mutations; 28.5% and 21%, respectively). Out of 14 patients with DNMT3A mutation, 9 patients had incomplete remission and only 5 achieved complete remission with no statistically significant association. Odds ratio of the response to induction therapy according to DNMT3A mutation status was 1.32 times higher to show incomplete remission than in wild-DNMT3A patients. Conclusion: DNMT3A mutation has high prevalence in AML Egyptian patients with non-statistically significant difference between mutated DNMT3A and wild type when related to the impact on remission rates after induction therapy.

Effect of FLT3-ITD with DNMT3A R882 double-mutation on the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

Objective: To investigate the impact of FLT3-ITD and DNMT3A R882 double mutations to the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods: FLT3-ITD, DNMT3A, C-kit, CEBPA, FLT3-TKD and NPM1 mutations were detected in 206 newly diagnosed AML patients by Sanger sequencing (M(3) and those received FLT3 inhibitor were excluded). Clinical data of AML patients were retrospectively analyzed to compare the prognosis of each gene mutation group. Results: ①Of 206 patients, 104 were male and 102 female with a median age of 38 (3-63) years, including 6 cases of M(0), 24 cases of M(1), 56 cases of M(2), 39 cases of M(4), 63 cases of M(5), 6 cases of M(6) and 12 unclassified cases. ②All 206 patients were divided into four groups according to the mutation gene at the time of diagnosis: FLT3-ITD(+) DNMT3A R882(+) group (group A), FLT3-ITD(+) DNMT3A R882(-) group (group B), FLT3-ITD(-) DNMT3A R882(+) group (group C) and FLT3-ITD(-) DNMT3A R882(-) groups (group D). Gender, leukocyte count at diagnosis, chromosome karyotype, the median age, FAB classification, disease status prior to transplantation, type of donor, conditioning regimen and GVHD were not significantly different between four groups (P>0.05). ③The 2-year cumulative recurrence rate (CIR) of group A was significantly higher than that of other groups [group A (72.2±2.6)%, group B (38.6±0.6)%, group C (36.8±1.6)%, group D (27.8±0.1)%, respectively, P<0.05], while the 2-year overall survival (OS) rate and 2-year leukocyte-free survival (LFS) rate were lower than those of other groups [group A (30.9±13.3)%, (11.3±10.2)%; group B (67.5±7.8)%, (47.9±8.4)%; group C (61.4±12.4)%, (56.8±12.5)%; group D (80.1±3.7)%, (79.7±3.6)%, respectively, P<0.05]. Conclusion: AML patients with FLT3-ITD and DNMT3A R882 double mutations had a very high CIR and low OS, LFS after transplantation.

Study on the Promoter Region-149C＞T and-579G＞T Polymorphism of DNMT3B with Genetic Susceptibility to Esophageal Cancer of Han Population in Jiangsu Suqian / 现代检验医学杂志

Objective To investigate the association of DNA methyltransfcrase 3B (DNMT3B) promoter region single nucleo tide polymorphism (SNP)-149C＞T and-579G＞T with genetic susceptibility of esophageal cancer (EC) of Han population in Jiangsu Suqian.Methods Gcnotypes of the-149C＞T and 579G＞T locus in DNMT3B promoter region were examinedby polymerase chain reaction with restriction fragment length polymorphism (PCR RFLP) in 246 esophageal cancer patients and 240 healthy controls.Results In-149C＞T site,the distributions of genotypes TT compared with CT in the EC group and the controls were no significant difference (x2 =0.089,P＞0.05).In-579G＞T site,the distributions of genotypes TT compared with GT+GG in the EC group and the controls were no significant difference (x2 =0.649,P＞0.05).When strat ified by age and gender,there was no significant difference in the EC group and the controls in both two sites (x2 =0.044～0.876,all P＞0.05).Conclusion The DNMT3B promoter region-149C＞T and-579G＞T were not associated with the genetic susceptibility of esophageal cancer.

The progress of pathogenesis and treatment in angioimmunoblastic T-cell lymphoma / 中国肿瘤临床

Angioimmunoblastic T-cell lymphoma (AITL) is one of the most common subtypes of peripheral T-cell lymphomas. It is a fol-licular T-helper-derived neoplasm, and characterized by the presence of some genetic alterations, such as mutations in TET2, DN-MT3A, IDH2, and RHOA. Anthracycline-containing regimens represent the most widely adopted first-line option; however, new biolog-ic agents should be designed and incorporated to improve treatment response. The elucidation of the specific roles of these genetic al-terations in AITL will facilitate the development of molecularly targeted therapies to treat this disease.

mRNA expression of DNA methyltransferase 2 and 3a in the epidermis of patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To investigate the expression of DNA methyltransferase 2 (DNMT2) and 3a (DNMT3a) in the epidermis of patients with psoriasis vulgaris.Methods Between March 2009 and December 2010,46 patients with psoriasis vulgaris were enrolled from the Department of Dermatology,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College,and the Department of Dermatology of Yixing People's Hospital,and 28 healthy controls were enrolled from the Department of Dermatologic Surgery,Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College.Real-time quantitative PCR was performed to determine the mRNA expression of DNMT2 and DNMT3a in the epidermis of the lesional and nonlesional skin of patients with psoriasis vulgaris,as well as in the epidermis of normal skin of the healthy controls.Results The mRNA expression of DNMT2 (expressed as 2-△△Ct) in the lesional skin,non-lesional skin of the patients and normal skin of the healthy controls was 0.62 ± 0.02,0.36-± 0.05 and 0.15 ± 0.11,respectively.The mRNA expression of DNMT2 was significantly higher in the lesional skin than in the non-lesional skin of the patients (t =6.23,P ＜ 0.01),and higher in the non-lesional skin of the patients than in the normal skin of the healthy controls (t =7.33,P ＜ 0.01).Additionally,the mRNA expression of DNMT3a was significantly higher in the lesional skin (0.85 ± 0.03) than in the non-lesional skin (0.43 ± 0.04) of the patients (t =5.66,P ＜ 0.01),and higher in the non-lesiona] skin of the patients than in the normal skin of healthy controls (0.18 ± 0.09,t =8.62,P ＜ 0.01).Conclusion Both DNMT2 and DNMT3a mRNA were abnormally expressed in the epidermis of patients with psoriasis vulgaris.

Effect of FLT3-ITD with DNMT3A R882 double-mutation on the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

Objective@#To investigate the impact of FLT3-ITD and DNMT3A R882 double mutations to the prognosis of acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT).@*Methods@#FLT3-ITD, DNMT3A, C-kit, CEBPA, FLT3-TKD and NPM1 mutations were detected in 206 newly diagnosed AML patients by Sanger sequencing (M3 and those received FLT3 inhibitor were excluded). Clinical data of AML patients were retrospectively analyzed to compare the prognosis of each gene mutation group.@*Results@#①Of 206 patients, 104 were male and 102 female with a median age of 38 (3-63) years, including 6 cases of M0, 24 cases of M1, 56 cases of M2, 39 cases of M4, 63 cases of M5, 6 cases of M6 and 12 unclassified cases. ②All 206 patients were divided into four groups according to the mutation gene at the time of diagnosis: FLT3-ITD+ DNMT3A R882+ group (group A), FLT3-ITD+ DNMT3A R882- group (group B), FLT3-ITD- DNMT3A R882+ group (group C) and FLT3-ITD- DNMT3A R882- groups (group D). Gender, leukocyte count at diagnosis, chromosome karyotype, the median age, FAB classification, disease status prior to transplantation, type of donor, conditioning regimen and GVHD were not significantly different between four groups (P>0.05). ③The 2-year cumulative recurrence rate (CIR) of group A was significantly higher than that of other groups [group A (72.2±2.6)%, group B (38.6±0.6)%, group C (36.8±1.6)%, group D (27.8±0.1)%, respectively, P<0.05], while the 2-year overall survival (OS) rate and 2-year leukocyte-free survival (LFS) rate were lower than those of other groups [group A (30.9±13.3)%, (11.3±10.2)%; group B (67.5±7.8)%, (47.9±8.4)%; group C (61.4±12.4)%, (56.8±12.5)%; group D (80.1±3.7)%, (79.7±3.6)%, respectively, P<0.05].@*Conclusion@#AML patients with FLT3-ITD and DNMT3A R882 double mutations had a very high CIR and low OS, LFS after transplantation.

DNA methyltransferase 3A regulates the expressions of collagens during the activation of cardiac fibroblasts / 医学研究生学报

Objective The mechanisms of methylation acting on myocardial fibrosis are not yet clear at present. The aim of this study was to investigate the role of DNA methyltransferase 3A (DNMT3A) in regulating the expressions of collagens during the activation of cardiac fibroblasts.Methods Cardiac fibroblasts were obtained from 50 neonatal mice and divided into three groups: blank control, DNMT3A overexpression plasmid (mDNMT3A-pEGFP-C3) and small interference DNMT3A siRNA. The contents of collagens in the cell supernatant were detected by ELISA. The mRNA and protein expressions of type I collagen (Col Ⅰ), type Ⅲ collagen (Col Ⅲ) and DNMT3A in the cardiac fibroblasts were determined by real-time quantitative PCR and Western blot respectively and the proliferative activity of the cardiac fibroblasts measured by CCK8 assay.Results The contents of Col I and Col Ⅲ in the cell supernatant were significantly increased in the DNMT3A overexpression plasmid group but decreased in the DNMT3A siRNA group as compared with those in the blank control (P<0.05). The expressions of Col Ⅰ, Col Ⅲ and DNMT3A were remarkably higher in the DNMT3A overexpression plasmid group but lower in the DNMT3A siRNA group than in the blank control (P<0.05). The cell activity was markedly higher in the DNMT3A overexpression plasmid group than in the empty vector plasmid and control groups (2.087±0.317 vs 1.063±0.223 and 1.082±0.207, P<0.05) but lower in the DNMT3A siRNA group (0.463±0.087) than in the latter two (P<0.05).Conclusion DNMT3A can increase the proliferation and activation of cardiac fibroblasts, upregulate the expressions of collagens and thus promote myocardial fibrosis.

The challenge for allogeneic hematopoietic stem cell transplantation in acute leukemia/ myelodyplastic syndrom with TP53, TET2 or DNMT3A gene mutations / 天津医药

@#Recently, much gene mutations have been detected in patients with acute leukemia or myelodysplastic syndrome (MDS) using next-generation sequencing (NSG) technology. Some of them are proved to be important prognostic markers. It has been showed that TP53, TET2 or DNMT3A gene mutations are associated with poor prognosis in acute leukemia or MDS patients. The prognosis of these patients is poor with short remission and survival. Allogeneic hematopoietic stem cell transplantation is the only way to cure these patients. However, the outcomes after transplantation are inferior to those in patients without these mutations. The hypomethylating agents or immune targeting therapy might improve their prognosis when combined with the present strategies. Here, the impact of TP53, TET2 and DNMT3A gene mutations on the prognosis after chemotherapy or transplantation is reviewed.

Expression of TET2 and DNMT3A in peripheral T cell lymphoma and their significance / 临床与实验病理学杂志

Purpose To investigate the expression of TET2 and DNMT3A in patients with peripheral T cell lymphoma (PTCL) and the relationship to immunophenotypes of PTCL.Methods Using a panel of immunohistochemical markers (CD3,CD4,CD10,BCL-6,CXCL-13,CD30,ALK),all cases of PTCLs were further divided into four groups,including angioimmunoblastic T cell lymphoma (AITL),peripheral T cell lymphoma,not otherwise specified (PTCL-NOS),anaplastic lymphoma kinase negative anaplastic large cell lymphoma (ALK-ALCL) and anaplastic lymphoma kinase positive anaplastic large cell lymphoma (ALK + ALCL).The expression of TET2 and DNMT3A in 89 cases of PTCL was detected by immunohistochemical analysis.Results 89 cases were divide into four subtypes,AITL (36/89),PTCL-NOS (26/89),ALKALCL (18/89),and ALK + ALCL (9/89).Immunohistochemistry staining revealed higher cytoplasmic expression of TET2 and DNMT3A in AITL than that of in PTCL-NOS and ALCL (P ＜ 0.05).And the nuclear expression of DNMT3A in patients with AITL was higher than that of PTCL-NOS and ALCL (P ＜ 0.05).The cytoplasmic expression of TET2 was positively related with both cytoplasmic and nuclear expression of DNMT3A in patients with AITL (P ＜ 0.05).Conclusion TET2 combined with DNMT3A could be used as markers in AITL diagnosis,which could provide new strategy for AITL diagnosis.