ABSTRACT
@#Objective: To investigate the relationship between drug resistance and expression of ABC-binding cassette subfamily B member 1 (ABCB1) as well as its promoter methylation in pancreatic cancer. Methods: Fifteen pairs of pancreatic cancerous tissues and corresponding para-cancerous tissues, which were pathologically verified in Fujian Cancer Hospital from August 2015 to August 2018, were collected for this study; in addition, 3 cases of normal pancreatic tissues and pancreatic cancer cell line SW1990 were also collected. Gemcitabine (GEM)-resistant human pancreatic cancer cell line SW1990/GZ was induced by intermittent concentration gradient multiplication method. The expression level ofABCB1 in SW1990 cells, SW1990/GZ cells, pancreatic cancer tissues and apara-cancerous tissues was detected by qPCR. Methylation of ABCB1 promoter region in SW1990 cells, SW1990/GZ cells and pancreatic cancerous tissues was determined by MSP-PCR. Results: Compared with SW1990 cells, the morphology of SW1990/GZ cells showed more vacuoles, more mitotic images, clumpy growth and increased drug resistance (P<0.05). ABCB1 expression in pancreatic cancer tissues was significantly higher than that in para-cancerous tissues (P<0.01). The expression of ABCB1 in SW1990 and SW1990/GZ cells was significantly higher than that in normal pancreatic tissues (P<0.05 or P<0.01), and the expression of ABCB1 in SW1990/GZ cells was higher than that in SW1990 cells (P<0.05). ABCB1 promoters in SW1990, SW1990/GZ cells and normal pancreatic tissues were hypomethylated. Rate of methylation in pancreatic can cerous tissues and normalpancreatic tissues was 6.7%(1/15) and 0.00%(0/3) respectively,and the difference was statistically insignificant (all P>0.05). Conclusion: Increased ABCB1 expression in pancreatic cancer tissues and cells is associated with drug resistance, but its gene expression does not depend on promoter methylation regulation.
ABSTRACT
OBJECTIVE@#To examine the clinical effects of Yisui Shengxue Granules () in the treatment of β-thalassemia and explore its mechanism on DNA methylation levels.@*METHODS@#A randomized placebo-controlled double-blinded trial was conducted. Forty patients with β-thalassemia were recruited and distributed randomly by envelope method into an experimental group and a control group, 20 patients in each group. The patients were given Yisui Shengxue Granules in the experimental group and placebo in the control group (12 g/bag three times a day) during a 3-month intervention. Before and after 1, 2, and 3 months of treatment, peripheral intravenous blood was sampled, and blood parameters such as hemoglobin (Hb), red blood cells (RBCs), reticulocytes (Ret), and fetal hemoglobin (HbF) were analyzed. Mononuclear cells from 5 patients, who showed an obvious treatment effect, were isolated by density gradient centrifugation. DNA methylation was analyzed using an Affymetrix USA GeneChip Human Promoter 1.0 Array and Input-promoter 1.0.@*RESULTS@#Compared with pre-treatment, there was an obvious increase in Hb and RBCs counts after 1, 2, and 3 months in the experiment group (P<0.01 or P<0.05). Meanwhile, HbF increased from the 2nd to the 3rd month (P<0.05). In the control group, Hb and RBCs showed no obvioas change. After 3-month treatment, DNA methylation results from 5 patients revealed that there were 24 hypomethylated genes and 3,685 hypermethylated genes compared with pre-treatment. Genes of insulin-like growth factor 1 receptor (IGF1R) and Janus kinase 3 (JAK3) revealed the most relations with other genes (degree: 21) and genes of 1-phosphatidylinositol-4, 5-bisphosphate phosphodiesterase gamma 2 (PLCG2) and mitogen-activated protein kinase 10 (MAPK10) showed a stronger intermediary role (betweenness centrality=0.04).@*CONCLUSIONS@#JAK3 and MAPK10 are two key genes in bone marrow and the lymphatic system, and JAK3 is likely to be related to hematopoietic cytokines in the process of early hematopoiesis. (Registration No. NCT01549080).
ABSTRACT
OBJECTIVE@#To investigate the changesof DNA methylation in histone deacetylases 4 gene (HDAC4) and its effectduring the trans-differentiation process of human mesenchymal stem cells (hMSCs) into sweat gland like cells (SGLCs).@*METHODS@#Selected cell lines of human mesenchymal stem cells (hMSCs) were cultured and expended , the third generation ofhMSCs and heat-shocked sweat gland cells were picked up, and were co-culturedwith adding inducible factor in the transwell chamber. The sweat gland like cells (SGLCs)in experiment group and the hMSCs in control group were collected, the changes of DNA methylation degree of CpG dinucleotide sitesin histone deacetylases 4 gene (HDAC4) promotor were detected by methylation specific PCR (MSP)andMaldi-TOF Mass Array. And then, the hMSCs in experiment group were treated with 5-aza-CdR (5-aza-2-deoxycytidine, 10 μmol/L), while the hMSCsin control group were culturedwith PBS at the same time. ThemRNA expressions of HDAC4 gene and carcino-embryonic antigen (CEA)gene in the two groups were measured by RT-PCR.@*RESULTS@#The methylation of HDAC4gene in hMSCs was in high level before induction, the methylation degreeof CpG dinucleotide sites located in cg2463009 was 0.901, and the methylation degree of HDAC4gene in SGLCs was markedly decreased by 37% after induction, which was 0.531. The methylationlevel of CpG dinucleotide sites located in cg14823429was changed from 0.687to 0.386 after induction. The mRNA expression of HDAC4 gene was upregulated in test group after treated with 5-aza-CdR for 48 hours, the mRNA expression of CEA gene related with transdifferentiation was enhanced too at the same term, there was significantly statistic difference compared with control group (<0.05).@*CONCLUSIONS@#Methylation of HDAC4 gene participates in the regulation of the trans-differentiation of hMSCs into sweet gland like cells.
Subject(s)
Humans , Azacitidine , Cell Differentiation , DNA Methylation , Histone Deacetylases , Mesenchymal Stem Cells , Repressor Proteins , Sweat GlandsABSTRACT
Depression is a common mental disease, which mainly presents with a state of low mood and aversion to activity. Epigenetics is to study the influence of structural modification of pre?transcriptional gene to gene function in chromatin level. Recent studies have found that epigenetic mechanisms including DNA methylation, histone modification, and non?coding RNA regulation play important roles in the pathogenesis of depression. This article reviews the research advances on epigenetic mechanisms of depression.
ABSTRACT
AIM:Toinvestigatetheassociationbetweenmethylationstatusofapoptosis-relatedgenesandche-mosensitivity in the lung adenocarcinoma cell line P 15.METHODS: Methylation-specific PCR was applied to detect the methylation status of p73, p14ARF, p16INK4a and bax genes of P15 cells in untreated control group and decitabine (DAC) treatment group.RT-PCR was used to detect the expression of p 73, bcl-xL, bad, bax, p14ARF and p16INK4a at mRNA level. Colony formation assay and cell growth inhibition assay were used to detect the sensitivity of P 15 cells to cis-diaminedichlo-roplatinum ( C-DDP) before and after DAC treatment .DAPI staining was used to determine the apoptosis of P 15 cells ex-posed to C-DDP before and after DAC treatment .RESULTS:p73, p16INK4a and bax were expressed in the methylation sta-tus.After DAC treatment, p16INK4a expression was decreased , and the expression of p73 and bax disappeared .The expres-sion of p73, p16INK4a and bax in the unmethylated status was weak , but the enhanced expression was observed following DAC treatment.After P15 cells were treated with DAC and C-DDP, the colony formation rate of the P15 cells was signifi-cantly decreased as compared with untreated control group .The apoptotic P15 cells in DAC+C-DDP treatment group were significantly higher than those in untreated control group (P<0.05).CONCLUSION:After treated with DAC, the sensi-tivity of P15 cells to C-DDP is increased due to the activation of silenced pro-apoptotic genes .DAC and C-DDP synergisti-cally promote tumor cell apoptosis .They have significant anti-tumor effect .