ABSTRACT
Wnt signaling are critical pathway involved in organ development, tumorigenesis, and cancer progression. WNT7A, a member of the Wnt family, remains poorly understood in terms of its role and the underlying molecular mechanisms it entails in head and neck squamous cell carcinoma (HNSCC). According to the Cancer Genome Atlas (TCGA), transcriptome sequencing data of HNSCC, the expression level of WNT7A in tumors was found to be higher than in adjacent normal tissues, which was validated using Real-time RT-PCR and immunohistochemistry. Unexpectedly, overexpression of WNT7A did not activate the canonical Wnt-β-catenin pathway in HNSCC. Instead, our findings suggested that WNT7A potentially activated the FZD7/JAK1/STAT3 signaling pathway, leading to enhanced cell proliferation, self-renewal, and resistance to apoptosis. Furthermore, in a patient-derived xenograft (PDX) tumor model, high expression of WNT7A and phosphorylated STAT3 was observed, which positively correlated with tumor progression. These findings underscore the significance of WNT7A in HNSCC progression and propose the targeting of key molecules within the FZD7/JAK1/STAT3 pathway as a promising strategy for precise treatment of HNSCC.
Subject(s)
Animals , Humans , Squamous Cell Carcinoma of Head and Neck , Carcinogenesis/genetics , Cell Transformation, Neoplastic , Wnt Signaling Pathway , Disease Models, Animal , Head and Neck Neoplasms/genetics , Wnt Proteins , Frizzled Receptors/genetics , Janus Kinase 1 , STAT3 Transcription FactorABSTRACT
Objective To assess the effects of hepatitis B virus(HBV) large envelope protein (LHB) on the apoptosis of HepG2 cells and explore the possible mechanism by proteomic approaches.Methods LHB gene was cloned into pShuttle-IRES-hrGFP-1,and the recombinant adenovirus either barboring LHB(Ad-LHB) or empty vector(Ad-GFP) were separately generated.Annexin V-FITC apoptosis detection kit,JC-1 mitochondrial membrane potential assay kit and propidium iodide(PI) staining kit were employed combined with flow cytometry to detect the apoptotic cells infected with Ad-LHB or control of Ad-GFP.The cellular proteins were collected after infection of HepG2 cells by Ad-LHBs or Ad-GFP,and a total of 600 μg proteins were submitted to two-dimensional gel electrophoresis(2-DE) and stained with R350.The gel images were captured by ImageScanner Ⅱ Imaging System,the differentially expressed proteins were identified by ImageMaster 2D Platinum analysis software and picked up by Ettan Spot Picker.After enzyme digestion,the protein samples were analyzed by MALDI-TOF-TOF MS.Results HepG2 cells infected with Ad-LHB were much more prone to apoptosis.There were thirty nine differentially expressed proteins were determined by 2-DE between HepG2 cells infected with Ad-LHB and Ad-GFP,and they were identified ultimately and categorized into thirty three kinds of proteins by MALDI-TOF-TOF MS.Among these proteins,nine were found to be closely related to cell apoptosis,in which CAPN2,eIF3K and PPP2CB were higher expressed in Ad-LHB infected HepG2 cells,and SERPINH1,LASP1,PRDX1,DHRS2,LDHA and PS-MA4 were lower expressed in Ad-LHB infected HepG2 cells.Conclusion LHB could induce apoptosis of HepG2 cells,and several apoptosis-related proteins participated in this process.
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Objective To study the structure of spliced variants of hepatitis B virus (HBV) genomes and elucidate their potential pathogenicity. Methods Amplified the spliced variants of hepatitis B virus genomes by mean of PCR from the serum of the patients with chronic hepatitis B, sequenced and analysis the characteristics of such genomes. Results 10 different types of the spliced variants of hepatitis B virus genomes were obtained with the molecular weight ranging from 765 bp to 2039 bp. There were 6 splicing donor sites and 6 accepter sites, respectively in HBV genomes. All spliced variants showed one or more deletions in the regions coding for core, preS1, preS2 and surface protein, while retained thepathogenic X gene and cis elements which were essential for viral replication and packaging. Conclusions Spliced variants of hepatitis B virus genomes were commonly detected in the serum from chronic Hepatitis B patients, the characteristic structure of such variants implied that they might closely co-related with the pathogenicity of HBV.
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Objective To investigate the signal transduction pathway of arachidonic acid(AA) and prostaglandin E-2(PGE-2) synthesis in macrophage invaded by Toxoplasma gondii. Methods Synthesis of AA and PGE-2, expression of COX_2 mRNA and protein following stimulation infection by Toxoplasma gondii were evaluated in RAW264
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Objective To obtain protein of the major surface antigen P30 of Toxoplasma gondii by molecularcloning. Methods The gene of P30 (containing the whole P30 gene sequence, without the gene encoding signalpeptide) was obtained by polymerase chain reaction (PCR) using the primer designed according to the DNA sequence ofP30. The recombinant plasmid was constructed using EcoR Ⅰ, Xho Ⅰ and was then transformed into E. coli Top10. Thepositive clones were identified by restriction enzymes and DNA sequence analysis. The fusion protein was induced by IPTGand purified by affinity chromatography using ProBond~(TM) Resin (a kind of nickel-charged sepharose resin) and was identi-fied by SDS-PAGE and Western blotting. Results The products of PCR, cleavage and link reaction were same as ex-pected and the sequence of inserted fragment in the recombinant plasmid was same as reported except one synonymy muta-tion. A 58 kDa fusion protein was induced by IPTG and was purified by chromatography. Conclusion Fusion proteincontaining Toxoplasma gondii P30 was achieved and was provided as experiment material for further research.
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Objective To investigate the relationships between the expression of VEGF,its receptor KDR/flk-1,cNOS mRNA and angiogenesis,cell proliferation,metastasis in human hepatocellular carcinoma(HCC). Methods Immunohistochemical analysis using antibodies against VEGF and its kinase insert domain receptor (KDR) was carried out.cNOS mRNA expression in HCC,liver cirrhosis and normal liver tissue was observed by in situ hybridization.CD34 immunostaining was used to measure the microvascular density(MVD)and proliferative index was evaluated by Ki-67 immunostaining. Results The expressions of VEGF and KDR of HCC were significantly related to MVD,proliferation and metastasis of HCC(P