IL-27 promotes anti-tumor effect of NK92 cells by up-regulating expression of activated receptors and STATs phosphorylation / 中国肿瘤生物治疗杂志

@#Objective: To investigate the molecular and signal pathway mechanism of Interleukin-27 affecting the anti-tumor effect of NK92 cells. Methods: NK92 cells were cultured with different concentrations of IL-27 (10, 20, 30 and 60 ng/ml) for 24 hours. The cytotoxicity of NK92 cells to target cells was detected by LDH assay. The expressions of NKG2D, NKp30 and NKp46 on the surface of NK92 cells and the secretion of perforin and granzyme B were detected by Flow cytometry. The expression and phosphorylation level of STATs protein was detected by WB. The DU145 cell transplanted tumor model of prostatic carcinoma in NOD-PrkdcscidIl2rgem1/Smoc mice was established and treated with the combination of NK92 cells and IL-27 to evaluate their anti-tumor efficacy. Results: IL-27 at concentrations of 10, 20 and 30 ng/ml could significantly increase the cytotoxicity of NK92 cells to target cells, and 30 ng/ml exerted the best effect (P<0.05 or P<0.01). 30 ng/ml IL-27 could significantly promote the expressions of NKG2D, NKp30 and NKp46 on surface of NK92 cells, as well as elevate the secretion of perforin (all P<0.05), but didn’t affect the secretion of granzyme B (P>0.05); moreover, it also up-regulated the phosphorylation of STAT1, STAT3 and STAT5 protein (all P<0.01). The combined treatment of IL-27 and NK92 cells obviously extended the survival time of tumor-bearing mice (P<0.05). Conclusions: IL-27 can promote the cytotoxicity of NK92 cells against solid tumor cells and blood tumor cells by promoting expressions of NKG2D, NKp30 and NKp46 on the surface of NK92 cells and the secretion of perforin, which might be related with the phosphorylation of STAT1, STAT3 and STAT5 in JAKSTAT pathway.

AKT/mTOR/STATs signaling pathway regulates various immune cells that mediate MS / 医学研究生学报

AKT/mTOR/STATs signaling pathway not only plays an important role in tumor growth, but also in the regulation of immune system. Activated AKT promotes the activation of downstream signaling pathways mTORC1 and mTORC2 through phosphorylation. mTOR is currently being considered as an important regulator of immune system and plays an important role in regulating the function and metabolism of various immune cells. Multiple sclerosis (MS) is an autoimmune deficiency disease whose pathogenesis has not been fully elucidated. This article focuses on the regulation of AKT/mTOR/STATs signaling pathways in various immune cells such as macrophage M1/M2 polarization, B cell proliferation and differentiation, helper T lymphocyte (Th cell) proliferation and differentiation, and Treg cell proliferation and differentiation, which would be helpful to illustrate the role of the AKT/mTOR/STATs signaling pathway in mediating the regulation of immune cells in MS.

Analysis of the Relevance between t-BHP Induced Apoptosis and STATs mRNA Expression in HEI-OC1 Cells / 听力学及言语疾病杂志

Objective To study the biological effects of (Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription (STATS) of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations (0,20,40,60μM) of(Tert-Butyl hydroperoxide,T-BHP) built the apoptotic HEI-OC1 cells model while 0 μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60 μM) were 3.9％,7.4％,32.0％,and 91.2％,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STATS mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly (F=5 534.302,P＜0.01;F=146.038,P＜0.01;F=685.929,P＜0.01;F=516.11,P＜ 0.01).No statistically significant differences were found among the poisoning concentration groups (P＞0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly (F=1 259.148,P＜ 0.01).The STAT2 mRNA expression level in 20 μM contamination group decreased while 40,60 μM contamination group increased significantly (P＜0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.

Biological functions and related diseases of STATs / 中国药科大学学报

@#The signal transducer and activator of transcription(STAT)proteins, which can transmit signals from extracellular to nucleus, play important roles in biological processes. The abnormal activation of STAT3 signaling pathway usually results in many malignant diseases such as tumors(leukemias, lymphomas, breast cancer and lung cancer), inflammation or immune and rheumatoid arthritis. Numerous studies have demonstrated that the activation of STAT3, which means the phosphorylation of the residue 705, can inhibit apoptosis, induce cell proliferation and angiogenesis and finally lead to malignant diseases mentioned above. In this review, the biological functions of STAT family proteins and the related diseases of STAT3 are introduced, and the inhibitors of STAT3 are summarized simply. Further development of STATs is also proposed.

Study on the correlation between the polymorphism of STATS gene and asthma / 中国医师杂志

Objective To investigate the correlation rs2272087 polymorphism of STATS gene and asthma.Methods The polymerase chain reaction PCR- SBT technique was used to determine rs2272087 polymorphism in asthma and control group.Results The genotype of AA,AG and GG of rs2272087 were 0.600,0.412 and 0.167 in asthma group,and 0.430,0.200,0.367 in control group,respectively.The frequency of allele A and G was 0.903 and 0.344 in asthma group,and 0.656,0.970 in control group,respectively.There was significant difference in two groups(x2 =9.40,P ＜0.01 ;x2 =11.58,P ＜0.01 ).Conclusions The rs2272087 polymorphism of STAT5 gene may be an important candidate gene for asthma.

Establishment and functional analysis of CHO cells with stable expression of hGHR gene and its mutants / 基础医学与临床

Objective To eonstruct three eukaryotic expression vectors containing wilde-type hGHR gene and its mutants(hGHR-E42K and hGHR-H56R) related to congenital growth hormone insensitivity, then check their expression in CHO cells. Methods With the available PUC-hGHR vector, two mutate hGHR genes (hGHR-E42K and hGHR-H56R) were obtained through mutagenesis. Then three recombinants were cloned into eukaryotic expression vectors pcDNA3.1/zeo(+) with restriction enzymatic reactions.Then with Lipofectamine2000, we trans-fected expression vectors to CHO cells and screened the stably expressed CHO cells by Zeocin. RT-PCR and/or Western blotting were used to examine hGHR and STAT5-P. Results After sequencing, two mutations were introduced to hGHR, three eukaryotic expression vectors were identified. The transfected CHO cells expressed vd-hGHR or its mutants. Compared with hGH-wt, two mutate cells (E42K and H56R) had decreased phosphorylated STAT5 levels. Conclusion Three CHO cells which stably expresses wide type hGHR and its mutants were successfully established, E42K and H56R partly interfered the phosphorylation of STAT5.

Induction of differentiation by diallyl disulfide through inhibition of JAK1/STAT3 in human leukemia HL-60 cells / 中国药理学通报

Aim To investigate JAKs/STATs signal transduction change in HL-60 cells differentiation induced by diallyl disulfide(DADS)and molecular mechanism regulating the differentiation.Methods After incubation of HL-60 cells with DADS or AG490(50 ?mol?L -1),the cell differentiation indexes were observed by cytomorphology, NBT reduction ability assay,cell myeloid differentiation antigen CD11b by flow cytometry. Kinase activity of JAKs/STATs was tested by western-blotting and expressions of nucleus transcription genes stats,c-myc,c-fos,c-jun were detected by immumocyte chemistry method.Results Cell differentiation index changes indicated that HL-60 cells were induced differentiation toward granulocytic lineage by DADS, Western blot test demonstrated that constitive phosphorylation of Jak1,stat3 kinase was suppressed. Stat3,c-myc gene expression decreased and c-fos, c-jun gene expression increased in HL-60 cells treated with DADS through immunocyte chemistry.Conclusion Inhibition of phosphative Jak1, Stat3 was involved in HL-60 cells differentiation induced by DADS, its molecular mechanism might be related to modulation of gene expression associated proliferation and differentiation,and inhibition of DNA systhesis, induction differentiation.