As one of the major risks for urolithiasis, hyperoxaluria can be caused by genetic defect or dietary intake. And high oxalate induced renal epithelial cells injury is related to oxidative stress and mitochondrial dysfunction. Here, we investigated whether MitoTEMPO, a mitochondria‐targeted antioxidant, could protect against oxalate mediated injury in NRK‐52E cells via inhibiting mitochondrial dysfunction and modulating oxidative stress. MitoSOX Red was used to determine mitochondrial ROS (mtROS) production. Mitochondrial membrane potential (Δψm) and quantification of ATP synthesis were measured to evaluate mitochondrial function. The protein expression of Nox4, Nox2, and p22 was also detected to explore the effect of oxalate and MitoTEMPO on NADPH oxidase. Our results revealed that pretreatment with MitoTEMPO significantly inhibited oxalate induced lactate dehydrogenase (LDH) and malondialdehyde (MDA) release and decreased oxalate induced mtROS generation. Further, MitoTEMPO pretreatment restored disruption of Δψm and decreased ATP synthesis mediated by oxalate. In addition, MitoTEMPO altered the protein expression of Nox4 and p22 and decreased the protein expression of IL‐6 and osteopontin (OPN) induced by oxalate. We concluded that MitoTEMPO may be a new candidate to protect against oxalate induced kidney injury as well as urolithiasis.

The molecular events leading to nephrolithiasis are extremely complex. Previous studies demonstrated that calcium and transforming growth factor-β1 (TGF-β1) may participate in the pathogenesis of stone formation, but the explicit mechanism has not been defined. Using a self-created genetic hypercalciuric stone-forming (GHS) rat model, we observed that the increased level of serous/uric TGF-β1 and elevated intracellular calcium in primary renal tubular epithelial cells (PRECs) was associated with nephrolithiasis progression in vivo. In the setting of high calcium plus high TGF-β1 in vitro, PRECs showed great potential epithelial to mesenchymal transition (EMT) progression and osteochondral differentiation properties, representing the multifarious increased mesenchymal and osteochondral phenotypes (Zeb1, Snail1, Col2A1, OPN, Sox9, Runx2) and decreased epithelial phenotypes (E-cadherin, CK19) bythe detection of mRNAs and corresponding proteins. Moreover, TGF-β-dependent Wnt11 knockdown and L-type Ca²⁺ channel blocker could greatly reverse EMT progression and osteochondral differentiation in PRECs. TGF-β1 alone could effectively promote EMT, but it had no effect on osteochondral differentiation in NRK cells (Rat kidney epithelial cell line). Stimulation with Ca²⁺ alone did not accelerate differentiation of NRK. Co-incubation of extracellular Ca²⁺ and TGF-β1 synergistically promotes EMT and osteochondral differentiation in NRK control cells. Our data supplied a novel view that the pathogenesis of calcium stone development may be associated with synergic effects of TGF-β1 and Ca²⁺, which promote EMT and osteochondral differentiation via Wnt11 and the L-type calcium channel.

Background. Bladder cancer is one of the most common malignancies of the urinary system with an unfavorable prognosis. More and more studies have suggested that lipid metabolism could influence the progression and treatment of tumors. However, there are few studies exploring the relationship between lipid metabolism and bladder cancer. This study aimed to explore the roles that lipid metabolism-related genes play in patients with bladder cancer. Methods. TCGA_BLCA cohort and GSE13507 cohort were included in this study, and transcriptional and somatic mutation profiles of 309 lipid metabolism-related genes were analyzed to discover the critical lipid metabolism-related genes in the incurrence and progression of bladder cancer. Furthermore, the TCGA_BLCA cohort was randomly divided into training set and validation set, and the GSE13507 cohort was served as an external independent validation set. We performed the LASSO regression and multivariate Cox regression in training set to develop a prognostic signature and further verified this signature in TCGA_BLCA validation set and GSE13507 external validation set. Finally, we systematically investigated the association between this signature and tumor microenvironment, drug response, and potential functions and then verified the differential expression status of signature genes in the protein level by immunohistochemistry. Results. A novel 6-lipidmetabolism-related gene signature was identified and validated, and this risk score model could predict the prognosis of patients with bladder cancer. In addition, the prognostic model was tightly related to immune cell infiltration and tumor mutation burden. Gene set variation analysis (GSVA) and gene set enrichment analysis (GSEA) showed that mTOR signaling pathway, G2M checkpoint, fatty acid metabolism, and hypoxia were enriched in patients in the high-risk score groups. Furthermore, 3 therapies specific for bladder cancer patients in different risk scores were identified. Conclusions. In conclusion, we investigated the lipid metabolism-related genes in bladder cancer through comprehensive bioinformatic analysis. A novel 6-gene signature associated with lipid metabolism for predicting the outcomes of patients with bladder cancer was conducted and validated. Furthermore, the risk score model could be utilized to indicate the choice of therapy in bladder cancer.

The present study aimed to identify the characteristics and cross‑talk between transforming growth factor β1 (TGF‑β1) and calcium ions in nephrolithiasis patients with idiopathic hypercalciuria (IH) in order to elucidate the potential mechanisms underlying changes in cell phenotype induced by bone‑associated factors and their influence on renal nephrolithiasis formation. Blood samples from a total of 29 nephrolithiasis patients with IH, 29 renal stone patients without IH and 29 healthy age‑matched normal controls were subjected to quantification of peripheral serum TGF‑β1, osteopontin (OPN) and bone morphogenetic protein 2 (BMP2) using ELISA. This was followed by detection of BMP2, OPN and 1,25‑dihydroxyvitamin D3 receptor (VDR) mRNA and protein levels in primary renal epithelial cells (PRECs) of IH and HK‑2 human proximal tubular cell lines (control) using reverse transcription quantitative polymerase chain reaction (RT‑qPCR) and western blot analyses. The mRNA expression levels of BMP2, OPN and VDR in PRECs and HK‑2 were evaluated following stimulation with various concentrations of TGF‑β1 (0.5, 2.0 and 5.0 ng/ml), Ca2+ (0.5, 1.5 and 2.5 mM) or TGF‑β1 and Ca2+ combined using RT‑qPCR, respectively. TGF‑β1, BMP2 and OPN expression levels in patients with IH were all significantly higher than those in the control group. The mRNA and protein expression levels of BMP2 and VDR were significantly higher in PRECs than those in HK‑2 cells. Following incubation with TGF‑β1 and/or Ca2+, the mRNA expression levels of BMP2, OPN and VDR in PRECs increased in a dose‑dependent manner; however, no significant differences were observed in HK‑2 cells with increasing TGF‑β1 dosage. Co‑incubation with TGF‑β1 and Ca2+ in PRECs and HK‑2 cell lines resulted in similar effects and the expression of BMP2, OPN and VDR mRNA increased in a time‑dependent manner. In conclusion, the results of the present study demonstrated that TGF‑β1 regulated the expression of BMP2, OPN and VDR in PRECs, but not in HK‑2 cells. Co‑incubation with TGF‑β1 and Ca2+ significantly increased the expression levels of bone‑associated factors in PRECs and HK‑2 cells, which suggested that this process may be partially responsible for the pathogenesis of calcium stone development, and also associated with bone formation and the TGF‑β1‑induced epithelial to mesenchymal transition.

Background. Epithelial-mesenchymal transition (EMT) plays a vital role in tumor metastasis and drug resistance. It has been reported that EMT is regulated by several long noncoding RNAs (lncRNAs). We aimed to identify EMT-related lncRNAs and develop an EMT-related lncRNA prognostic signature in kidney renal clear cell carcinoma (KIRC). Materials and Methods. In total, 530 ccRCC patients with 611 transcriptome profiles were included in this study. We first identified differentially expressed EMT-related lncRNAs. Then, all the samples with transcriptional data and clinical survival information were randomly split into training/test sets at a ratio of 1:1. Accordingly, we further developed a twelve differentially expressed EMT-related lncRNA prognostic signature in the training set. Following this, risk analysis, survival analysis, subgroup analysis, and the construction of the ROC curves were applied to verify the efficacy of the signature in the training set, test set, and all patients. Besides, we further investigated the differential immune infiltration, immune checkpoint expression, and immune-related functions between high-risk patients. Finally, we explored the different drug responses to targeted therapy (sunitinib and sorafenib) and immunotherapy (anti-PD1 and anti-CTLA4). Results. A twelve differentially expressed EMT-related lncRNA prognostic signature performed superior in predicting the overall survival of KIRC patients. High-risk patients were observed with a significantly higher immune checkpoint expression and showed better responses to the targeted therapy and immunotherapy. Conclusions. Our study demonstrates that the twelve differentially expressed EMT-related lncRNA prognostic signature could act as an efficient prognostic indicator for KIRC, which also contributes to the decision-making of the further treatment.

Background. TP53 mutation is a common mutation gene in uterine corpus endometrial carcinoma (UCEC), and the TP53 signaling pathway plays an essential role in the tumorigenesis, progression, and immune infiltration in UCEC. We aimed to discover TP53 pathway‐related lncRNAs in UCEC. Materials and methods. 528 UCEC patients with 587 transcriptional profiles were enrolled in this study. We first investigated the differential status of TP53 signaling pathway between tumor and normal tissues by GSEA analysis, then identified TP53 pathway‐related lncRNAs, accordingly establishing a nine TP53 pathway related to the lncRNA signature in the training set and verified this signature in the test set. Besides, the interaction network was constructed; the immune infiltration, drug response to cisplatin and paclitaxel, and mutation atlas were investigated. Finally, we performed a subgroup analysis to check the universality of this signature. Results. A nine TP53 pathway‐related lncRNA prognostic signature was constructed and verified superior accuracy in predicting the overall survival of UCEC patients. Besides, high‐risk patients showed a poor prognosis, but they were more sensitive to the cisplatin and paclitaxel. Notably, M2 macrophages were higher infiltrated in high‐risk patients, and TP53 showed a significantly higher mutation in high‐risk patients than low‐risk patients. Conclusions. We constructed and verified a nine TP53 pathway‐related lncRNA prognostic signature in UCEC, which also contributes to the decision‐making of the chemotherapy.