Astragalus polysaccharide inhibits the development of urothelial carcinoma by activating AMPK signaling to induce BENC1-xCT complex formation.

In recent years, the incidence of urothelial carcinoma (UC) has been high in men. The aim of this study was to investigate whether astragalus polysaccharide (APS) could inhibit the development of UC and the specific molecular mechanism. Our data showed that APS inhibited the proliferation of UC cells in a dose-dependent manner, and APS reduced the migratory capacity of RT4 and T24 cells. Further studies revealed that the ferroptosis inhibitor ferrostatin-1 (Fer-1) reversed APS-induced cell death, intracellular Fe2+ and malondialdehyde (MDA) accumulation, and lipid peroxidation product deposition. The Western blot and immunofluorescence results showed that APS significantly inhibited the expression of glutathione peroxidase 4 (GPX4) but did not alter the protein level of solute carrier family 7 member 11 (xCT, SLC7A11). Further analysis revealed that APS reduced the activity of xCT in RT4 and T24 cells. Moreover, APS significantly increased the phosphorylation levels of protein kinase AMP-activated catalytic subunit alpha 1 (AMPK) and BECN1 in RT4 and T24 cells, which induced the formation of the BECN1-xCT complex. However, when AMPK was silenced in RT4 and T24 cells, APS-induced ferroptosis was reversed to some extent, indicating that APS-mediated ferroptosis involves AMPK signaling. Moreover, APS has been shown to inhibit tumor growth in nude mice in vivo. In summary, our study demonstrated for the first time that APS could promote the formation of the BECN1-xCT complex in UC cells by activating AMPK/BECN1 signaling, which inhibited the activity of xCT to reduce GPX4 expression, thereby inducing ferroptosis and ultimately inhibiting UC progression.

Search for the optimized and key nephrometry elements combination in retroperitoneal laparoscopic partial nephrectomy: A retrospective study.

Background: The nephrometry scoring system plays a key role in the preoperative evaluation of partial nephrectomy, and scoring systems based on anatomical characteristics have high similarity in scoring elements. Currently, there is little research on scoring systems related to retroperitoneal laparoscopic partial nephrectomy, and there is a lack of research on the combination of scoring elements, which requires further investigation. Methods: We retrospectively analyzed the clinical records of 107 patients who underwent retroperitoneal laparoscopic partial nephrectomy conducted by a single operator at a single center. The score and scoring elements were generated based on imaging. The scoring elements of each scoring system and all combinations of two to five elements were extracted. The predictive ability of different score combinations was evaluated by AUC value, and the key parameters of the score were found by taking the intersection. A nomogram was constructed and evaluated. Results: We observed that with an increase in scoring elements, the strongest combination of elements did not significantly increase the predictive ability of warm ischemia time (P>0.05), postoperative complications (P>0.05), and trifecta achievement (P>0.05). The combination of the maximum tumor diameter and the distance between tumor and collecting system or renal sinus had a good comprehensive predictive ability, and there is no significant difference with the traditional score (P>0.05). The nomogram generated according to this combination has an excellent prediction ability for predicting whether obtain trifecta of partial nephrectomy. Conclusions: Within the range of two to five elements, the critical degree of elements is more important than the number of elements. The maximum tumor diameter and the distance between the tumor and the collecting system or renal sinus was the key element of the prediction ability.

A Novel M7G-Related MicroRNAs Risk Signature Predicts the Prognosis and Tumor Microenvironment of Kidney Renal Clear Cell Carcinoma.

Background: M7G modification is extremely vital for the development of many cancers, especially tumor immunity. M7G modification is a novel functional regulator of miRNA, and the researches on m7G-related miRNAs in kidney renal clear cell carcinoma (KIRC) are still insufficient. This research aims to establish a risk signature on the foundation of m7G-associated miRNAs, which can precisely forecast the prognosis of KIRC patients. Methods: Transcriptome data and clinical data used in this study come from The Cancer Genome Atlas database. Our team utilized univariable Cox, Lasso and multivariable Cox analyses to construct a m7G-associated miRNAs risk signature that can forecast the prognosis of KIRC patients. Kaplan-Meier method, time-dependent receiver operating characteristic (ROC) curve, and the independent analysis of risk signatures were employed to verify the predictability and accuracy of the risk signature. Subsequently, based on CIBERSORT, ESTIMATE and ssGSEA algorithms, we speculated the potential impact of the proposed risk signature on tumor immune microenvironment. Ultimately, by virtue of the risk signature and tumor immunity, the hub genes affecting the prognosis of KIRC patients were screened out. Results: Our team established and verified a prognostic signature comprising 7 m7G-associated miRNAs (miR-342-3p, miR-221-3p, miR-222-3p, miR-1277-3p, miR-6718-5p, miR-1251-5p, and miR-486-5p). The results of the Kaplan-Meier survival analysis revealed that the prognosis of KIRC sufferers in the high-risk group was often unsatisfactory. The accuracy of the prediction ability of the risk signature was verified by calculating the area under the ROC curve. Univariate-multivariate Cox analyses further showed that this risk signature could be utilized as an independent prognosis-related biomarker for KIRC sufferers. The results of the immune analysis revealed that remarkable diversities existed in immune status and tumor microenvironment between high-risk and low-risk groups. On the foundation of the proposed risk signature and other clinical factors, a nomogram was established to quantitatively forecast the survival of KIRC sufferers at 1, 3 and 5 years. Conclusion: Based on m7G-related miRNAs, a risk signature was successfully constructed, which could precisely forecast the prognosis of sufferers and guide personalized immunotherapy for KIRC patients.

A novel defined risk signature based on pyroptosis-related genes can predict the prognosis of prostate cancer.

BACKGROUND: Pyroptosis can not only inhibit the occurrence and development of tumors but also develop a microenvironment conducive to cancer growth. However, pyroptosis research in prostate cancer (PCa) has rarely been reported. METHODS: The expression profile and corresponding clinical data were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Patients were divided into different clusters using consensus clustering analysis, and differential genes were obtained. We developed and validated a prognostic biomarker for biochemical recurrence (BCR) of PCa using univariate Cox analysis, Lasso-Cox analysis, Kaplan-Meier (K-M) survival analysis, and time-dependent receiver operating characteristics (ROC) curves. RESULTS: The expression levels of most pyroptosis-related genes (PRGs) are different not only between normal and tumor tissues but also between different clusters. Cluster 2 patients have a better prognosis than cluster 1 patients, and there are significant differences in immune cell content and biological pathway between them. Based on the classification of different clusters, we constructed an eight genes signature that can independently predict the progression-free survival (PFS) rate of a patient, and this signature was validated using a GEO data set (GSE70769). Finally, we established a nomogram model with good accuracy. CONCLUSIONS: In this study, PRGs were used as the starting point and based on the expression profile and clinical data, a prognostic signature with a high predictive value for biochemical recurrence (BCR) following radical prostatectomy (RP) was finally constructed, and the relationship between pyroptosis, immune microenvironment, and PCa was explored, providing important clues for future research on pyroptosis and immunity.

Erdafitinib exerts the anticancer effect on urothelial carcinoma via induction of authophagy.

Erdafitinib has recently been approved for the treatment of adult patients with locally advanced or metastatic urothelial carcinoma (UC). The current study aimed to evaluate whether erdafitinib inhibits the proliferation in UC via inducing authophagy. Our data showed that erdafitinib demonstrated strong toxicity for the T24 and UMUC6 cell lines. Flow cytometry analysis showed that erdafitinib increased the proportion of G0/G1 phase in both T24 and UMUC6 cells. Additionally, Annexin V staining indicated that erdafitinib enhanced the apoptosis of UC cells. Meanwhile, the expression of apoptosis-related proteins was significantly elevated after treatment with erdafitinib. Transwell assay showed that erdafitinib significantly reduced T24 and UMUC6 cell migration and invasion. More importantly, western blot assay showed that erdafitinib induced the expression of autophagy-related proteins. Besides, GFP-LC3 transfection assay and electron microscopy examination showed that erdafitinib could partially diminish 3-methyladenine (3-MA) induced impairment of autophagy in UC cells. In summary, for the first time we showed novel data that erdafitinib inhibited UC cell malignant proliferation via inducing cell autophagy.

The Effects of Orientin on Proliferation and Apoptosis of T24 Human Bladder Carcinoma Cells Occurs Through the Inhibition of Nuclear Factor-kappaB and the Hedgehog Signaling Pathway.

BACKGROUND Orientin is a flavone isolated from medicinal plants used in traditional Chinese medicine (TCM), which suppresses the growth of cancer cells in vitro. The effects of orientin in bladder cancer cells remains unknown. This study aimed to investigate the effect of orientin on proliferation and apoptosis of T24 human transitional cell bladder carcinoma cells in vitro in the presence of an agonist and an inhibitor of nuclear factor-kappaB (NF-kappaB). MATERIAL AND METHODS T24 cells were cultured and divided into four study groups: an untreated control group; a group treated with 100 µM orientin; a group treated with 100 µM orientin with NF-kappaB agonist, phorbol 12-myristate 13-acetate (PMA); and a group treated with 100 µM orientin and the NF-kappaB inhibitor, IkappaBalpha. The MTT assay was performed to assess cell viability, and flow cytometry evaluated the cell cycle. The expression of proteins in the Hedgehog signaling pathway and inflammatory cytokines were determined by Western blot and enzyme-linked immunosorbent assay (ELISA). RESULTS Orientin inhibited the proliferation of T24 cells, caused cell cycle arrest, reduced cell viability, and inhibited the expression of inflammatory mediators. Treatment of T24 cells with orientin inhibited the expression of NF-kappaB and components of the Hedgehog signaling pathway, and the NF-kappaB agonist, PMA, reversed these effects. CONCLUSIONS Treatment of T24 human bladder carcinoma cells in vitro with orientin inhibited cell proliferation and promoted cell apoptosis by suppressing the Hedgehog signaling pathway and NF-kappaB.

Diagnostic Value of Circulating miR-324 for Prostate Cancer.

BACKGROUND: To investigate the diagnostic value of serum miR-324 in patients with prostate cancer (PC). METHODS: Blood samples from 50 patients with prostate cancer, 30 patients with benign prostatic hyperplasia (BPH), and 20 healthy controls were collected and quantified by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The relationship between serum miR-324 level and Gleason classification and TNM staging of prostate cancer was analyzed. ROC curve was used to analyze the value of miR-324 in early diagnosis of prostate cancer. RESULTS: The expression of miR-324 in prostate cancer group was significantly higher than that in the BPH group and normal control group (all p < 0.01). Serum miR-324 was associated with the Gleason score and tumor stage (all p < 0.01). Serum miR-324 was positively correlated with PSA (r = 0.673, p = 0.000); ROC curve analysis showed that the area under the ROC curve of miR-324 (all p < 0.01) (AUC) was 0.911 (95% CI: 0.855 - 0.966, p = 0.000). When the relative expression level of miR-324 was 3.35, the sensitivity, specificity, positive predictive value, and negative predictive values were 86.0%, 82.0%, 82.7%, and 85.4%, respectively. CONCLUSIONS: The increased expression of serum miR-324 in patients with prostate cancer may be a potential new biomarker for the diagnosis of prostate cancer, which is helpful for the differentiation between PC and BPH.

High Novel Oncogene with Kinase-Domain (NOK) Gene Expression is Associated with the Progression of Renal Cell Carcinoma.

BACKGROUND: To study the expression levels of the NOK (novel oncogene with kinase-domain) gene in renal cell carcinoma and its association with the progression of this cancer. METHODS: In this study, immunohistochemistry (IHC) and Western blot analyses were applied to investigate the NOK expression level in RCC and adjacent normal renal tissue samples. MTT, colony formation, and migration assays were also utilized to evaluate the role of NOK in RCC cell lines. RESULTS: Knocked-down expression of NOK in an RCC cell line (786-0) suppressed cellular proliferation and migration by restraining the activation of AKT and ERK. We found that the expression level of NOK was significantly higher in RCC tissues than in their adjacent tissues, and more importantly, overexpression of NOK was evidently correlated with the tumor TNM stage and Fuhrman grade (p < 0.001). A high level of NOK was also associated with poor overall survival (p < 0.05) and disease-free survival (p < 0.05) by Kaplan-Meier analysis. CONCLUSIONS: NOK expression increased in RCC and was significantly correlated with TNM stage, Fuhrman grade, poor overall survival, poor disease-free survival, metastasis, and proliferation in RCC cells by regulating the activation of AKT and ERK, suggesting that NOK may play important roles as a positive regulator to RCC.

The l58Val/Met polymorphism of catechol-O-methyl transferase gene and prostate cancer risk: a meta-analysis.

The association between COMT Val158Met polymorphism and prostate cancer has been evaluated. However, the results of these studies on the association remain conflicting. To derive a more precise estimation of the relationship, a meta-analysis was performed. A comprehensive search was conducted to identify the eligible studies of COMT Val158Met polymorphism and prostate cancer risk. Summary odds ratios (OR) and 95 % confidence interval (CI) for COMT Val158Met polymorphism and prostate cancer were calculated. Statistical analysis was performed with the software program Review Manage (Version 5.0) and Stata (Version 12.0). Six case-control studies, totally 4,118 persons including 2,143 cases and 1,975 controls, met the included criteria and thus were selected. Our analysis suggested that Val158Met polymorphism was associated with prostate cancer risk in overall population. Collectively, the results of the present study suggest that significant associations of COMT Val158Met polymorphisms with prostate cancer were observed (for additive model: OR = 1.068, 95 % CI = 1.002-1.138, P (heterogeneity) = 0.363, P = 0.043; for dominant model: OR = 1.266, 95 % CI = 1.057-1.517, P (heterogeneity) = 0.000, P = 0.011; for recessive model: OR = 1.050, 95 % CI = 0.961-1.146, P (heterogeneity) = 0.558, P = 0.279; and Val allele versus Met allele OR = 0.932, 95 % CI = 0.894-0.971, P (heterogeneity) = 0.272, P = 0.001). In the subgroup analysis, we detected no significant association between the COMT 158 Val/Met genotype and prostate cancer risk in Caucasian and Asian populations, while the contrary result for additive model (OR = 2.43, 95 % CI = 1.08-5.43, P (heterogeneity) = 0.04, P = 0.03) in Asian populations. The result of this meta-analysis suggests that COMT l58Val/Met polymorphism might be contributed to the overall prostate cancer risk.

[V89L polymorphism of the testosterone 5-alpha-reductase II gene and prognostic factors of prostate cancer].

OBJECTIVE: To investigate the association of V89L polymorphism of the SRD5A2 gene with the prognostic factors of prostate cancer (PCa). METHODS: We identified the V89L polymorphic sites of the SRD5A2 gene after Rsa-1 restriction enzyme digestion, observed the distribution of V89L (VV, VL and LL) polymorphism in 112 PCa and 89 benign prostate hyperplasia (BPH) patients, and determined the association of V89L polymorphism with the age, free PSA (fPSA), total PSA (tPSA), fPSA/tPSA ratio, tumor stage and Gleason score of the PCa patients. RESULTS: No statistically significant differences were found in the V89L polymorphism-induced genetic risk frequencies between the PCa and BPH groups (chi2 = 3. 606, df = 2, P = 0. 165), nor any significant correlation between the genotypes of VV and VL + LL and the differences in the fPSA, tPSA, fPSA/tPSA ratio, tumor stage, Gleason score and age of the PCa patients. VV and VL + LL showed no obvious association with the prognostic factors of PCa. CONCLUSION: V89L polymorphism is not related with the prognosis of PCa, but may be indirectly associated with its risk.