HMG20A was identified as a key enhancer driver associated with DNA damage repair in oral squamous cell carcinomas.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the main type of oral cancer. Disturbing DNA repair is an invaluable way to improve the effectiveness of tumor treatment. Here, we aimed to explore the key enhancer drivers associated with DNA damage repair in OSCC cells. METHODS: Gene Set Enrichment Analysis (GSEA), Gene Set Variation Analysis (GSVA) and Kaplan-Meier analysis were applied to explore the relationship among DNA repair-related genes expression and clinical phenotypes based on The Cancer Genome Atlas (TCGA) database. HOMER software and Integrative Genomics Viewer were applied to identify and visualize enhancers using GSE120634. Toolkit for Cistrome Data Browser was applied to predict transcription factors. Human Protein Atlas Database was used to analyze the protein levels of transcription factors in OSCC and control tissues. Seventy-two OSCC patients were included in this study. qRT-PCR was used to detect transcription factor expression in OSCC and adjacent control tissues collected in this study. qRT-PCR and ChIP-qPCR were used to verify the binding of transcription factors to enhancers, and regulation of target genes transcription. Transcription factor knockdown and control cells were treated with cisplatin. CCK8 was used to detect cell viability and proliferation. Western blotting was implemented to detect the levels of DNA repair-related proteins. Transwell assay was used to detect cell invasion. RESULTS: DNA repair was positively associated with the OSCC metastatic phenotype. Patients in the cluster with high expression of DNA repair-related genes had a worse prognosis and a higher proportion of advanced stage, low-differentiation, alcohol consumption and smoking compared to the cluster with low DNA repair-related gene expression. Seventeen metastasis-specific enhancer-controlled upregulated DNA repair-related genes, with the top two upregulated genes being ADRM1 26 S proteasome ubiquitin receptor (ADRM1) and solute carrier family 12 member 7 (SLC12A7) were screened. High mobility group 20 A (HMG20A) was the key prognostic enhancer driver regulating metastasis-specific DNA repair-related genes, with higher expression in OSCC tissues than normal control tissues, and higher expression in metastatic OSCC tissues than non-metastatic OSCC tissues. HMG20A bound to the metastasis-specific enhancers of ADRM1 and SLC12A7, thereby promoting ADRM1 and SLC12A7 expression. Knockdown of HMG20A enhanced cisplatin sensitivity of cells, and inhibited OSCC cells from repairing DNA damage caused by cisplatin, as well as proliferation and invasion of OSCC cells. CONCLUSION: HMG20A was identified as the key prognostic enhancer driver regulating DNA repair in OSCC cells, providing a new therapeutic target for OSCC.

Pharmacokinetic Study on the Triptolide in Normal Rats and Adjuvant Arthritis Model Rats in vivo / 中国药房

OBJECTIVE:To study the pharmacokinetics of triptolide in Tripterygium glycosides tablet in normal rats and adju-vant arthritis model rats in vivo,and provide reference for clinical rational drug use. METHODS:12 SD rats were randomly divid-ed into normal group and model group,6 in each group. Model group was subcutaneously injected complete Freund's adjuvant 0.1 mL to induce adjuvant arthritis model,normal group was subcutaneously injected the same volume of saline. After 14 d model-ing,2 groups were given Tripterygium glycosides tablet suspension 96 mg/kg intragastrically,the blood sample of eyes 0.4 mL were respectively taken before and 10,30,45,60,90,120,150,180,240,300,420 min after administration. The plasma con-centration of triptolide was determined by HPLC,the pharmacokinetic parameters were calculated by DAS 2.0 software,and the parameters were compared. RESULTS:The pharmacokinetic parameters of triptolide in normal group were cmax of(1.139±0.114)μg/mL,tmax of(2.167±0.606)h,t1/2α of(5.500±3.610)h,AUC0-7 h of(5.052±0.371)μg·h/mL,MRT0-7 h of(3.224±0.119)h, and CL of(11.616±2.986)mL/h;and those in model group were cmax of(0.916±0.103)μg/mL,tmax of(3.083±0.801)h,t1/2αof(5.593±1.795)h,AUC0-7 h of(4.707±0.347)μg·h/mL,MRT0-7 h of(3.429±0.139)h,and CL of(11.246±2.638) mL/h. Compared with normal group,cmax in model group was significantly decreased,tmax and MRT0-7 h were significantly prolonged(P<0.05). CONCLUSIONS:Adjuvant arthritis can affect the pharmacokinetics of triptolide in rats in vivo,and promote its absorption and removal.

Study on the up-regulation of expression of B7 molecules and immunogenicity of acute leukemia cells induced by cytarabine / 白血病·淋巴瘤

Objective To investigate the effects of cytarabine(Ara-C) on expression of B7 molecules and immunogenicity of acute leukemia (AL) cells. Methods The expression of B7 molecules on fresh AL ceils and on the Ara-C exposed leukemia ceils was detected by FACS cytometer. B7 mRNA in Ara-C treated HL-60 cells were detected by reverse RT-PCR. The stimulation of proliferation of allogeneic PBMC by Ara-C treated HL-60 cells was detected by MTT method. Results B7-2 was weakly expressed in four and positive in three, whereas BT-1 was positive in only one of fourty AL patiens. Ara-C significantly enhanced B7 molecules expression on AL cells. Ara-C could induce higher expression of B7 mRNAs on HL-60 cells.Ara-C treated HL-60 cells could stimulate PBMC proliferation and promote their IFN -γ production.Conclusion Fresh AL cells express low level of B7 molecules. Ara-C enhances the B7 molecules expression on AL cells. The Ara-C treated leukemia cells can significantly stimulate the proliferation of allogeneic PBMC and induce their secretion of IFN-γ.

Repair potential of heat-treated rat bone marrow mesenchymal stem cells in infarcted myocardium / 中华器官移植杂志

Objective To study the effects of heat shock treatment of rat bone marrow mesenehymal stem cells(MSCs),the apoptosis ratio of treated-cells under low serum condition and the treated-cells transplantation on left ventricular function in rats with myocardiaIinfarction.Methods MSC8 were heat-treated under 42℃for 30 min,then the heat shock protein-70(HSP-70)was detected bv Western blot.The apoptosis ratio of heat-treated MSCs under low serum condition was tested by Annexin kit.The treated-MSCs labeled with Dil were transplanted into infarcted myocardium and 8 weeks later,the cardiac function of rats in each group was evaluated by echocardiography and cardiac catheterization.Results The immunophenotype of heat-treated MSCs did not vary,Western blot confirmed a higher level expression of HSP-70 in the treated-MSCs group as compared with that in the control group.The early apoptosis ratio was lower in treated-MSCs measured by flow cytometry with annexin staining than that of MSCs when cultured with low serum medium.After 8 weeks,LVEF,LVSP,+dp/dtmax,and-dp/dtmax were significantly higher,and the LVEDP was significantly lowar in heat-treated MSCs transplantation group than that in the control group.Conclusions Heat shock pretreatment of MSCs enhances the tolerance of MSCs to low serum medium,whereas does not lcad to the change of the cell immunophenotype.Transplantation of heattreated MSCs might improve the cardiac function in a rat myocardialinfarction model.

The impact of cell doses in graft on acute graft-versus-host disease following HLA-identical sibling ailogeneic peripheral blood hematopoietic stem cell transplantation / 中华器官移植杂志

Objective To explore the influence of mononuclear cells (MNC), CD34+ cells, CD3+ , CD3+ CD4+ , CD3+ CD8+ , CD4+ CD25+ T cells, CD3- CD16+ CD56+ natural killer cells (NKs), and dendritic cells (DCs) doses in graft on acute graft-versus-host disease (aGVHD) following HLA-identical sibling allogeneic peripheral blood hematopoietic stem cell transplantation (allo-PBSCT).Methods Sixty-five patients receiving HLA-identical sibling allo-PBSCT were studied.The number of CD34+, CD3+, CD3+ CD4+, and CD3+ CD8+ T cells in the graft was counted by fluorescence-activated cell sorting (FACS).The number of CD4+ CD25+ T cells, CD3 CD16+ CD56+ NKs, and DCs in the graft was also measured by FACS in 31 patients among above-mentioned 65 patients.The doses of each kind of cells in the graft were calculated according to per kilogram of recipients body weight.The patients were divided into high or low dose groups according to whether or not more than or equal to median of MNC, CD34+, CD3+, CD3+ CD4+, CD3+CD8+, CD4+ CD25+, CD3 CD16+ CD56+ or DC cell doses, respectively.Acute GVHD was analyzed between two groups.Results The frequency of the cumulative incidence of grade Ⅱ～Ⅳ aGVHD was increased in CD3+ CD4+ and CD3+ CD8+ T cells high dose groups as compared with correspondingly low dose groups, but the difference had no statistically significant difference (P = 0.089 and 0.098, respectively).Recipients in CD4 + CD25 + T cells high dose group had significantly reduced cumulative incidence of grade Ⅲ～Ⅳ aGVHD as compared with those in correspondingly low dose group (P< 0.05).The cumulative incidence of total aGVHD was significantly higher in DC1 high dose group than in correspondingly low dose group (P<0.05) and the cumulative incidence of grade Ⅱ～Ⅳ aGVHD was also higher in high dose group, but the difference had no statistically significant difference (P = 0.069).There was no significant difference in cumulative incidence of total and grade Ⅱ～Ⅳ aGVHD between MNC, CD34+ , CD3+, NK or DC2 high dose groups and correspondingly low dose groups (P>0.05, respectively).Conclusion Recipients in DC1 high dose group have significantly increased cumulative incidence of total aGVHD, but those in CD4+ CD25+ T cells high dose group have significantly reduced cumulative incidence of grade Ⅲ～Ⅳ aGVHD.