METTL3 and IGF2BP1-mediated m6A modification of ZHX2 promotes tumor property of renal cell carcinoma.

INTRODUCTION: Renal cell carcinoma (RCC) is a common type of kidney cancer with limited treatment options and a high mortality rate. Therefore, it is essential to understand the role and mechanism of key genes in RCC development and progression. This study aimed to analyze the role of zinc fingers and homeoboxes 2 (ZHX2) in RCC and the underlying mechanism. METHODS: RNA expression was analyzed by quantitative real-time polymerase chain reaction, while protein expression was analyzed by western blotting assay and immunohistochemistry assay. Cell viability was evaluated using CCK-8 assay, and cell proliferation was assessed by EdU assay. The rate of cell apoptosis was quantified by flow cytometry. Transwell assays were conducted to analyze cell migration and invasion. The sphere formation assay was performed to assess the formation of microspheres. Additionally, m6A RNA immunoprecipitation assay and RNA immunoprecipitation assay were utilized to investigate the relationship between ZHX2 and two proteins, methyltransferase like 3 (METTL3) and insulin like growth factor 2 mRNA binding protein 1 (IGF2BP1). The stability of ZHX2 mRNA was analyzed through the Actinomycin D assay. Furthermore, a xenograft mouse model assay was conducted to analyze the effect of ZHX2 overexpression and METTL3 silencing on RCC cell tumor properties in vivo. RESULTS: ZHX2 expression was upregulated in both RCC tissues and cells when compared with healthy renal tissues and human renal cortex proximal convoluted tubule epithelial cells. Depletion of ZHX2 inhibited RCC cell proliferation, migration, invasion and spheroid-forming capacity but promoted cell apoptosis. Moreover, it was found that METTL3 mediated m6A methylation of ZHX2 and IGF2BP1 also stabilized ZHX2 through m6A methylation modification. Furthermore, ZHX2 overexpression showed a potential for attenuating the effects induced by METTL3 silencing and counteracted the inhibitory effect of METTL3 depletion on tumor formation in vivo. CONCLUSION: METTL3 and IGF2BP1-mediated m6A modification of ZHX2 promoted RCC progression. The finding suggests that ZHX2 may serve as a potential therapeutic target in RCC, providing valuable insights for future clinical interventions.

Antimicrobial resistance and genetic relatedness of Salmonella serotypes isolated from food, asymptomatic carriers, and clinical cases in Shiyan, China.

Salmonella is a primary cause of foodborne diseases globally. Despite food contamination and clinical infections garnering substantial attention and research, asymptomatic Salmonella carriers, potential sources of infection, have been comparatively overlooked. In this study, we conducted a comparative analysis of serotype distribution, antimicrobial resistance phenotypes, and genetic profiles of archived Salmonella strains isolated from food (26), asymptomatic carriers (41), and clinical cases (47) in Shiyan City, China. Among the 114 Salmonella strains identified, representing 31 serotypes and 34 Sequence Types (STs), the most prevalent serovars included Typhimurium, Derby, Enteritidis, Thompson, and London, with the most predominant STs being ST11, ST40, ST26, ST34, and ST155. Antimicrobial resistance testing revealed that all strains were only sensitive to meropenem, with 74.6% showing antimicrobial resistance (AMR) and 53.5% demonstrating multidrug resistance (MDR). Strains resistant to five and six classes of antibiotics were the most common. Pearson's chi-square test showed no statistically significant difference in the occurrence of AMR (p = 0.105) or MDR (p = 0.326) among Salmonella isolates from the three sources. Our findings underscore associations and diversities among Salmonella strains isolated from food, asymptomatic carriers, and clinical patients, emphasizing the need for increased vigilance towards asymptomatic Salmonella carriers by authorities.

Expression of ß-adrenergic receptor subtypes in human normal and dilated ureter.

BACKGROUND: Ureter peristalsis is a basic physiological function regulated by myogenic and neurogenic factors. The distribution and function of ß-adrenergic receptors (ß-AR) in the human ureter remain unknown. The aim of this study was to investigate the expression of ß-AR subtypes in the normal and dilated human ureter. METHODS: The upper, middle, and lower segments of normal and dilated ureters were collected from patients undergoing surgery for carcinoma of the kidney and upper urinary tract and ureteral stenosis. The mucosa and muscular layers were separated. Expression of ß1-AR, ß2-AR, and ß3-AR mRNA and protein levels were detected by real-time PCR, western blot, and immunohistochemistry. RESULTS: In both mucosa and muscular layers, the mRNA and protein expressions of ß1-AR, ß2-AR, and ß3-AR were lower in the dilated ureter compared with the normal ureter. ß1-AR mRNA was significantly decreased (by 76.64%; P < 0.01) in the mucosa layer of the middle segment of the dilated ureter. ß1-AR and ß3-AR mRNA were significantly decreased (by 75.53 and 53.62%, respectively; P < 0.01) in the muscular layer of the lower segment of the dilated ureter. Similar findings were observed for protein expression. CONCLUSIONS: The downregulation of ß-ARs after ureter dilation, particularly for ß1-AR and ß3-AR in the muscular layer, suggests a potential compensatory mechanism involving increased contraction of the ureter to push urine through the obstruction. Thus, ß-ARs may be a potential target for treatment of ureter obstruction.