【Objective】 To investigate the effects of WM-3835, a histone acetyltransferase KAT7 (KAT7) inhibitor, on the proliferation and migration of bladder cancer cells and to explore the possible mechanism. 【Methods】 Human ureteral epithelial immortalized cell line SV-HUC-1, and bladder cancer cell lines UM-UC-3 and T24 were treated with different concentrations of WM-3835 (0, 10, 20, 30, 40 μmol/L). After 48 hours, the effects of WM-3835 on the proliferation, cell cycle distribution and migration of cells were detected with MTT assay, flow cytometry, scratch and Transwell assay, respectively. The expressions of cyclin D1 (cyclin D1), proliferating nuclear antigen (PCNA), matrix metalloproteinase 9 (MMP9) and neurocadherin (N-cadherin) were detected with Western blotting and real-time quantitative PCR. 【Results】 WM-3835 significantly inhibited the proli-feration of bladder cancer cells in a dose-dependent manner. After treatment with WM-3835, the cycle of UM-UC-3 and T24 cells were blocked in the G0/G1 phase, the proliferation was effectively inhibited, and the migration was significantly wea-kened. The expressions of cyclin-D1, PCNA, MMP9 and N-cadherin were down-regulated. 【Conclusion】 WM-3835 can inhibit the proliferation and migration of bladder cancer cells, and has the potential as a chemotherapeutic agent for bladder cancer.

Objective To investigate the expression levels and clinical significance of differentially expressed microRNAs(miRNAs)based on high-throughput sequencing in patients with left ventricular hypertrophy(LVH)and their possible regulatory mechanisms.Methods The miRNA sequencing was performed on plasma samples from 4 LVH patients and 4 healthy volunteers,and qRT-PCR validation was performed on plasma samples from 25 healthy volunteers and 35 LVH patients.The target genes of the obtained differentially expressed miRNA were predicted.The molecular function(MF),biological process(BP)and cellular components(CC)of these miRNAs were predicted by the Gene Ontology(GO)enrichment analysis,and their signal pathways were predicted by the Kyoto Encyclopedia of Genes and Genomes analysis.Meanwhile,the protein-protein interaction network was constructed.The diagnostic values of 2 differentially expressed miRNAs(hsa-miR-942-5p and hsa-miR-184)in LVH were analyzed by the area under the receiver operating characteristic(ROC)curve(AUCROC).Results A total of 945 differentially expressed miRNAs were identified,of which 1 and 9 were observed to be significantly upregulated and downregulated,respectively.The expression levels of hsa-miR-942-5p and hsa-miR-184 were verified by qRT-PCR to be significantly downregulated.The AUCROC of hsa-miR-942-5p and hsa-miR-184 were 0.694 6 and 0.880 4,respectively.Bioinformatics analysis revealed that the target genes were mainly enriched in the cell senescence,sphingolipid metabolism,and TGF-β,p53,and diabetic metabolism related signaling pathways.Conclusion The expression levels of hsa-miR-942-5p and hsa-miR-184 in plasma of LVH patients are significantly decreased,suggesting that they have good diagnostic potential.

c-Myc protein encoded by c-Myc (cellular-myelocytomatosis viral oncogene) gene regulates the related gene expression through the Wnt/β-catenin signaling pathway, and has received extensive attention in recent years. The purpose of this study was to express Helicoverpa armigera c-Myc gene (Ha-c-Myc) by using prokaryotic expression system, prepare the polyclonal antibody, examine the spatio-temporal expression profile of Ha-c-Myc, and investigate the possible function of Ha-c-Myc in regulating H. armigera sterol carrier protein-2 (SCP-2) gene expression. The Ha-c-Myc gene was amplified by PCR and cloned into a prokaryotic expression plasmid pET-32a(+). The recombinant plasmid pET-32a-Ha-c-Myc was transformed into Escherichia coli BL21. IPTG was used to induce the expression of the recombinant protein. Protein was purified by Ni²⁺-NTA column and used to immunize New Zealand rabbits for preparing the polyclonal antibody. The Ha-c-Myc expression levels in different developmental stages (egg, larva, prepupa, pupa, and adult) of H. armigera and different tissues (midgut, fat body, head, and epidermis) of the prepupa were determined by real-time quantitative reverse transcription PCR (qRT-PCR). Ha-c-Myc siRNA was synthesized and transfected into H. armigera Ha cells. The relative mRNA levels of Ha-c-Myc and HaSCP-2 in Ha cells were detected by qRT-PCR. Results showed that the pET-32a-Ha-c-Myc recombinant plasmid was constructed. The soluble Ha-c-Myc protein of about 65 kDa was expressed in E. coli. The polyclonal antibody was prepared. Western blotting analysis suggested that the antibody had high specificity. Enzyme linked immunosorbent assay (ELISA) showed that the titer of the antibody was high. Ha-c-Myc gene expressed at all developmental stages, with high levels in the early and late instars of larva, and the prepupal stage. Tissue expression profiles revealed that Ha-c-Myc expressed in various tissues of prepupa, with high expression level in the midgut, but low levels in the epidermis and fat body. RNAi results showed that the knockdown of Ha-c-Myc expression significantly affected transcription of HaSCP-2, leading to a 50% reduction in HaSCP-2 mRNA expression level. In conclusion, the Ha-c-Myc was expressed through a prokaryotic expression system, and the polyclonal anti-Ha-c-Myc antibody was obtained. Ha-c-Myc may promote the expression of HaSCP-2 and play an important role in the lipid metabolism of H. armigera. These results may facilitate further study on the potential role and function mechanism of Ha-c-Myc in H. armigera and provide experimental data for exploring new targets of green pesticides.
Animals ; Rabbits ; Escherichia coli/metabolism* ; Enzyme-Linked Immunosorbent Assay ; Moths/genetics* ; Blotting, Western ; Larva/genetics* ; Isoantibodies/metabolism* ; Antibody Specificity