Research progress of the mechanism for PD-1 molecule regulating pulmonary fibrosis and its research prospects in silicosis / 公共卫生与预防医学

Programmed death factor-1 (PD-1) is a promising target molecule for clinical tumor immunotherapy in recent years. Recent studies suggest that PD-1 and related signaling pathways (PI3K/AKT, JAK/STAT3, p38MAPK, ERK, etc.) played a key regulatory role in the process of pulmonary fibrosis. Silicosis is a systemic disease caused by inhalation of free silicon dioxide dust, which is mainly characterized by extensive pulmonary nodular fibrosis and seriously endangers the health of patients. Dissecting the role of PD-1 in the pathogenesis of silicosis may be of great significance in the mechanism research and clinical diagnosis and treatment of silicosis. This paper reviews the regulation of PD-1 molecule on related signaling pathways and its role in pulmonary fibrosis, and looks forward to the potential application of these mechanistic studies in silicosis research.

Inhibitory effect of LZJ541, a novel small molecule inhibitor of STAT3, on the proliferation of hepatocellular carcinoma cells / 药学学报

Signal transducer and activator of transcription 3 (STAT3) is an important regulatory factor of cell proliferation and metastasis, involved in the occurrence and development of a variety of malignant tumors, and it is one of the hot spots in the research of targeted anti-tumor drugs. Our group screened a novel benzobis (imidazole) structure small molecule compound LZJ541 through the screening model of Janus kinase (JAK)/STAT3 pathway inhibitors, which has definite STAT3 inhibitory activity. We examined the effect of LZJ541 on the proliferation of HepG2 and PC-3 cells by MTT assay in vitro, detected the effect of LZJ541 on the expression of STAT3-related proteins in HepG2 cells by Western blot, and measured the effect of LZJ541 on the apoptosis and cell cycle arrest of HepG2 cells via flow cytometry. The results indicated that LZJ541 significantly inhibited the activation of STAT3 signaling pathway and restrained the proliferation of HepG2 cells. Its half maximal inhibitory concentration (IC50) was 13.8 μmol·L-1, which was much lower than that of PC-3 cells (with low STAT3 expression, IC50: 41.99 μmol·L-1), LZJ541 can also inhibit the phosphorylation of STAT3 in HepG2 cells, thereby inducing apoptosis and cycle arrest and then exerting anti-tumor effects. In conclusion, LZJ541 has a certain anti-tumor effect in vitro, which provides an experimental basis for the development of new STAT3-targeted anti-tumor drugs around this kind of compounds.

Effect of anemoside B4 on proliferation, migration and apoptosis of ovarian cancer SKOV3 cells and its mechanism / 中华内分泌外科杂志

Objective:To investigate the effects of anemoside B4 on proliferation, apoptosis and migration of ovarian cancer SKOV3 cells through a variety of biological methods, and further to explore its mechanism.Methods:Ovarian cancer SKOV3 cells were cultured in vitro. CCK-8 method was used to detect the proliferation of SKOV3 cells treated with different concentrations of anemoside B4, and IC50 value was calculated. Flow cytometry was employed to detect the apoptosis of cells treated with IC50 concentration of anemoside B4 for different time length; Transwell method was used to detect the migration and invasion of cells treated with IC50 concentration of anemoside B4 for different time length. Western blot was used to detect changes in the expression of related proteins.Results:Anemoside B4 can effectively inhibit the proliferation of SKOV3 cells, showing a concentration-dependent IC50 value of 6.08±0.56 μM, and the inhibitory effect is stronger than the positive control drug cisplatin, with statistically significant difference (P<0.05) . Flow cytometry showed that anemoside B4 could induce SKOV3 cells apoptosis, reduce Bcl-xl expression, and up-regulate the expression of Bax, cleaved-caspase-3 and PARP. Compared with the group of 0 h, the difference was statistically significant (P<0.05) . Crocetin could down-regulate the expression of N-cadherin and up-regulate the expression of E-cadherin, thereby inhibiting the migration of SKOV3 cells. Anemoside B4 could inhibit the expression of JAK/STAT3 signaling pathway proteins.Conclusion:Anemoside B4 can effectively inhibit the proliferation of cervical cancer cells, and induce SKOV3 cell apoptosis by regulating the JAK/STAT3 signaling pathway to inhibit their migration. Crocetin has great potential in the research and development of ovarian cancer therapy.

Effects of IL-2 on extracellular matrix synthesis in retinal pigment epithelial cells / 西安交通大学学报(医学版)

Objective: To determine the effects of interleukin 2 (IL-2) on epithelial-mesenchymal transition (EMT) and extracellular matrix (ECM) synthesis in retinal pigment epithelial (RPE) cells. Methods: IL-2 of 10 μg/L was used to induce RPE cells. Real-time PCR and Western blot methods were used to detect the EMT marker α-smooth muscle actin (α-SMA), the extracellular matrix (ECM) markers fibronectin (Fn) and type I collagen (COL-1), transforming growth factor β2 (TGF-β2), and the activation of the JAK/STAT3 signaling pathway at corresponding time points. Furthermore, JAK/STAT3 signaling pathway was specifically blocked by WP1066, and the changes in α-SMA, COL-1, Fn and TGF-β2 mRNA and protein expressions were detected. Results: After induction by 10 μg/L of IL-2, the expressions of Fn, COL-1, TGF-β2 mRNA and protein as well as p-STAT3/STAT3 were significantly increased (P<0.05). This effect was correlated with the length of IL-2 treatment, while α-SMA mRNA and protein expression did not change significantly. JAK/STAT3 inhibitor WP1066 could effectively inhibit the expressions of Fn, COL-1 and TGF-β2 in IL-2-induced RPE cells. Conclusion: IL-2 promotes ECM synthesis and TGF-β2 expression in RPE cells via JAK/STAT3 signaling pathway, which may play an important role in proliferative vitreoretinopathy.

Cancer-Associated Fibroblasts Promote the Chemo-resistance in Gastric Cancer through Secreting IL-11 Targeting JAK/STAT3/Bcl2 Pathway / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Our aim was to detect the potential role of interleukin 11 (IL-11) in the development of chemo-resistance in gastric cancer and to reveal the mechanism involved in the process. MATERIALS AND METHODS: Here, we used flow cytometry to examine the percentage of cancer-associated-fibroblasts in tumor samples from chemo-resistant and -sensitive gastric cancer patients. Using MTT assay, we detected the cell viability under different conditions. Using quantitative real-time polymerase chain reaction and Western blotting, we determined the target expressions in mRNA and protein levels. We also performed immunohistochemistry and immunofluorescence to detect the target proteins under different conditions. Animal models were constructed to verify the potential role of IL-11 in chemo-resistant develop in vivo. RESULTS: Herein, we observed enriched cancer associated fibroblasts in drug resistant tumor tissues from gastric patients. Those fibroblasts facilitate the chemotherapeutic drugs resistance development through the secretion of IL-11, which activates the IL-11/IL-11R/gp130/JAK/STAT3 anti-apoptosis signaling pathway in gastric cancer cells. We found that the combination of chemotherapeutic drugs and JAK inhibitor overcomes the resistance and increases the survival of mice with gastric cancer xenografts. CONCLUSION: Ourresults demonstrated that IL-11 contributed to the obtain ofresistance to chemotherapy drugs through gp130/JAK/STAT3/Bcl2 pathway, and targeting the IL-11 signaling pathway induced by fibroblasts might be a promising strategy to overcome the multi-drugs resistant cancer in clinic.

JAK/STAT3SignalingPathwayIsInvolvedinPromotingtheProliferationand MigrationofBronchialSmoothMuscleCellsInducedbyInterleukin-17 / 中山大学学报(医学科学版)

@#：【Objective】Toinvestigatetheeffectsofinterleukin-17（IL-17）ontheproliferationandmigrationof bronchialsmoothmusclecells（BSMC）andtheroleofJAK/STAT3signalingpathwayinthisprocess.【Methods】BSMC weretreatedwithdifferentconcentrationsofIL-17fordifferenttimestodeterminethebestoftheexperimentalcondition. ThenMTTassaywasusedtodetectcellviability.CellproliferationstatesweredetectedbyBrdUstaining，andthecell cyclewasassessedbyPIstainingusingaflowcytometer.Transwellcellmigrationassaywasfurtherusedtodetectcell migrationability.TheexpressionofJAK，p-JAK，STAT3andp-STAT3inBSMCafterbeingtreatedwithIL-17was detectedbyWesternblotting.JAK/STAT3signalingpathwayspecificblockerAG490wasusedtoinvestigatetheroleof JAK/STAT3signalingpathwayinIL-17-inducedBSMCproliferationandmigration.TheeffectsofIL-17oncellproliferation， migration and JAK/STAT3 signaling pathway related protein expression were evaluated after blocking the JAK/STAT3 signaling.【Results】IL-17enhancedtheproliferation（P<0.05），promotedthecellcycletransitions（P<0.05）andsig⁃nificantlyincreasesthemigrationability（P<0.05）inBSMC.ThisprocesswasaccompaniedbytheenhancementofJAK/ STAT3signalingpathwayinBSMC（P<0.05）.InhibitionofJAK/STAT3signalingpathwayalleviatedBSMCproliferation andmigrationinducedbyIL-17（P<0.05） .【Conclusions】JAK/STAT3signalingpathwayparticipatesinthestimulation processofIL-17ontheproliferationandmigrationofBSMC.AG490inhibitstheenhancementofJAK/STAT3signaling pathwayinBSMCinducedbyinterleukin-17.

Effect and related mechanism on inflammatory response,proliferation and apoptosis of oxLDL-induced vascular smooth muscle cell of matrine / 中国免疫学杂志

Objective:To explore the effect and related mechanism on inflammatory response,proliferation and apoptosis of oxLDL-induced vascular smooth muscle cell of matrine.Methods:Theatherosclerotic model was conducted through treating human aort-icvascular smooth muscle cell with oxidized low density lipoprotein(oxLDL).Cell viability and proliferation was detected by CCK-8 as-say.The mRNA level of interleukin 1 beta(IL-1β),tumor necrosis factor alpha(TNF-α),IL-10 and IL-13 was tested by quantitative real-time reverse transcription PCR(qRT-PCR).Cell apoptosis was measured by flow cytometry.The expression of proliferation marker proteins antigen identified by monoclonal antibody(Ki-67) and proliferating cell nuclear antigen(PCNA),apoptosis marker proteins B cell lymphoma 2(Bcl-2) and Bcl-2-associated X protein(Bax),signal transducer and activator of transcription 3(STAT3) and signal transducer and activator of transcription 5(STAT5) was detected by Western blot.Results: Compared with control group,the mRNA level of IL-1β and TNF-α in model group was largely increased with decreased mRNA level of IL-10 and IL-13(P<0.01).Compared with model group,the mRNA level of IL-1β and TNF-α in treatment group was attenuated with enhancive mRNA level of IL-10 and IL-13(P<0.05).Cell proliferation and apoptosis in model group was higher than control group.Cell proliferation and apoptosis in treatment group was lower than model group(P<0.05).Compared with control group,the expression of Ki-67,PCNA and Bax in model group was augmented with decreased expression of Bcl-2(P<0.01).Compared with model group,the expression of Ki-67,PCNA and Bax in treatment group declined with elevated expression of Bcl-2(P<0.05).The expression of p-STAT3 and p-STAT5 in model group was higher than control group(P<0.01).The expression of p-STAT3 and p-STAT5 in treatment group was lower than model group(P<0.05).Compared with model group,the expression of Ki-67,PCNA and Bax in treatment group and model+Ruxolitinib group was decreased with enhancive expression of Bcl-2(P<0.05).The expression of Ki-67,PCNA and Bax in model+matrine+Ruxolitinib group was observably lower than model group and the expression of Bcl-2 in model+matrine+Ruxolitinib group was observably higher than model group(P<0.01).Compared with model group,the mRNA level of IL-1β and TNF-α in treatment group,model+Ruxolitinib group and model+matrine+Ruxolitinib group was attenuated with enhancive mRNA level of IL-10 and IL-13(P<0.05).Conclusion:Matrine represses inflammation,proliferation and apoptosis of vascular smooth muscle cell by inhibiting activation of JAK/STAT3 signal pathway in atherosclerosis.

Progress in research on JAK/STAT3 signaling pathway regulating tumor and related biological functions / 基础医学与临床

JAK/STAT3 signal pathway is an important intracellular signal transduction pathway. It plays important roles in cell apoptosis and proliferation by affecting the activity of downstream effector molecules,and also induces embryonic development,liver regeneration,glycolysis and inflammation,epithelial-mesenchymal transition and an-giogenesis and a series of biogenesis process,closely related to the development of human cancer.

Progress in research on JAK/STAT3 signaling pathway regulating tumor and related biological functions / 基础医学与临床

JAK/STAT3 signal pathway is an important intracellular signal transduction pathway. It plays important roles in cell apoptosis and proliferation by affecting the activity of downstream effector molecules,and also induces embryonic development,liver regeneration,glycolysis and inflammation,epithelial-mesenchymal transition and an-giogenesis and a series of biogenesis process,closely related to the development of human cancer.

Advances in Study on JAK/STAT3 Signaling Pathway and its Role in Gastrointestinal Diseases / 胃肠病学

JAK/STAT3 signaling pathway exists in various organs and tissues and mediates multiple biological processes including cell proliferation,differentiation,migration,apoptosis and immunoregulation.Recently,it has been revealed that this pathway plays an important role in gastrointestinal diseases,promoting tumor growth,angiogenesis and inhibiting apoptosis in gastric and colorectal cancers,and being implicated in the pathogenesis of inflammatory bowel disease.Inhibitors targeting JAK/STAT3 pathway showed promising outcome in some disease models.In this review article,the advances in study of abovementioned issues were summarized.

ITF increases the transcriptional activity of ITF promoter via the JAK-STAT3 signal transduction pathway / 中南大学学报(医学版)

Objective: To investigate the effect ofintestinal trefoil factor (ITF) on the transcriptional activity of ITF promoter and to explore the regulatory mechanismofJanus kinase/signal transducersand activators of transcription(JAK/STAT) on ITF promoter. Methods: The 5' flanking sequence of the ITF gene was cloned from human whole blood genomic DNA by PCR. ITF promoter fragment was cloned and inserted into the pGL3-Basic vector to construct recombinant vector. ITF promoter vector was stimulated with ITF at various concentrations and the luciferase activity was measured. The JAK-STAT3 signal transductionpathway was then blocked by a speciifc inhibitor AG490 to determine the signal pathway involved in ITF promoter activity. Results: Restriction endonuclease analysis and DNA sequencing confirmed that the recombinant plasmid, containing ITF promoter, was constructed successfully. After transient transfection, the activity of ITF promoter was increased signiifcantly in the presence of ITF (P<0.05). Blockage of the JAK-STAT3 signal transduction pathway with AG490 signiifcantly reduced the ITF promoter activity (P<0.05). Conclusion: ITF increases the transcriptional activity of ITF promoter via the JAK-STAT3 signal transduction pathway.

STAT3 mutations correlated with hyper-IgE syndrome lead to blockage of IL-6/STAT3 signalling pathway.

Of all the causes identified for the disease hyper-immunoglobulinemia E syndrome (HIES), a homozygous mutation in tyrosine kinase2 (TYK2) and heterozygous mutations in STAT3 are implicated the defects in Jak/STAT signalling pathway in the pathogenesis of HIES. Mutations of STAT3 have been frequently clinically identified in autosomaldominant (AD) HIES patients’ cells, and therefore, the genotype of STAT3 has been associated with the phenotype of HIES. Here, we conducted studies on the functional loss of the seven specific STAT3 mutations correlated with ADHIES. Using STAT3-null human colon carcinoma cell line A4 cells, we generated seven mutants of STAT3 bearing single mutations clinically identified in AD-HIES patients’ cells and studied the functional loss of these mutants in IL- 6-Jak/STAT3 signalling pathway. Our results show that five STAT3 mutants bearing mutations in the DNA-binding domain maintain the phosphorylation of Tyr705 and the ability of dimerization while the other two with mutations in SH2 domain are devoid of the phosphorylation of Try705 and abrogate the dimerization in response to IL-6. The phosphorylation of Ser727 in these mutants shows diversity in response to IL-6. These mutations eventually converge on the abnormalities of the IL-6/Gp130/Jak2-mediated STAT3 transactivation on target genes, indicative of the dysregulation of JAK/STAT signalling present in HIES.

Arsenic trioxide induced JAK/STAT3 pathway inhibition in myeloma cell lines / 上海交通大学学报（医学版）

Objective To explore the possible relationship between alteration of cell cycle and JAK/STAT3 signal transduction pathway inhibition induced by arsenic trioxide (As_2O_3,) in myeloma cell lines U266 and RPMI8226 in vitro. Methods Multiple myeloma cell lines U266 and RPMI8226 were used as in vitro models. The influence of AS_2O_3 on myeloma cells were evaluated by MTT assay and flow cytometry. Meanwhile, methylation specific PCR and Western blotting were employed to detect the methylation status of gene SOCS-1 and protein expression level of P-STAT3 in these cells after AS_2O_3 treatment. Results AS_2O_3 significantly inhibited the growth of U266 and RPMI8226 cells in a dose-dependent manner. Furthermore, cell cycle was arrested at G0/G1 phase with inhibition of protein expression level of P-STAT3 and SOCS-1 gene demethylation after exposure to As_2O_3 for 72 h( r = 0. 85, P < 0.05). Conclusion AS_2O_3 could induce the alteration of cell cycle which might be related to JAK/STAT3 signal transduction pathway inhibition and SOCS-1 demethylation in myeloma cell lines. The study puts forward a new idea of AS_2 O_3 treatment in multiple myeloma.