FTO mediated ERBB2 demethylation promotes tumor progression in esophageal squamous cell carcinoma cells.

N6-methyladenosine (m6A) is the most prevalent and internal modification that occurs in the messenger RNAs of eukaryotes. However, knowledge of the impact of these modifications on gene expression regulation remains limited. By using the in vitro MeRIP-seq and RNA-seq assays, we discovered that the mRNA demethylase FTO was significantly up-regulated in esophageal squamous cell carcinoma (ESCC) tissues and cells. Knockdown of FTO drastically suppressed the proliferation, migration, and invasion of ESCC cells. Furthermore, by using transcriptome-wide m6A-seq and RNA-seq assays, we identified ERBB2 is the target of FTO, which acts in concert in ESCC tumorigenesis and metastasis. Moreover, loss and gain functional studies suggested that the m6A reader YTHDF1 stabilizes ERBB2 mRNA via decoding the m6A modification. All these results uncovered a new signaling cascade, including FTO, YTHDF1, and ERBB2, which finely regulates the ESCC progression.

Extraperitoneal laparoscopic simple prostatectomy with urethra preservation using urethral initiation as the entry point: a practical approach for the treatment of benign prostatic obstruction.

PURPOSE: The present study introduces a modified surgical procedure, extraperitoneal laparoscopic simple prostatectomy (LSP) with urethra preservation using urethral initiation as the entry point, and evaluates its feasibility, safety, and efficacy in the treatment of benign prostatic obstruction (BPO). MATERIALS AND METHODS: A total of 50 patients with BPO underwent modified LSP from January 2018 to December 2020. The main surgical procedures performed were as follows: transversely incision of prostate surgical capsule at the urethral initiation; creating of the subcapsular plane and the space between urethra and adenoma; removal of lobes with preservation of urethra followed by suturing of capsule. Preoperative, perioperative, follow-up parameters, and complications were recorded and analyzed. RESULTS: Operative time was (106.34 ± 28.00) min and intraoperative blood loss was (98.80 ± 130.58) ml. Continuous bladder irrigation (CBI) was not performed routinely, catheterization duration was (5.26 ± 2.99) days, and postoperative hospital stay was (5.42 ± 1.62) days. Significant improvements were observed in functional outcomes, whereas no retrograde ejaculation, urinary incontinence, and urethral stricture occurred. Urethral rupture was not significantly influenced by operative time, intraoperative blood loss, and prostate volume. However, it prolonged CBI duration, drainage tube retention time, catheterization duration, and postoperative hospital stay. Operative time decreased with an increase in the number of cases, and the surgeon achieved proficiency level after handling 21-25 cases. CONCLUSION: Extraperitoneal LSP with urethra preservation using urethral initiation as the entry point is a feasible, repeatable, safe, and effective surgical procedure, which is suitable for treating BPO.

A New Functional Gene, Zinc Finger Protein 485 (ZNF485), is Involved in Bladder Cancer Proliferation.

BACKGROUND: Bladder cancer is the second most common urological cancer worldwide, with low early diagnosis and high mortality. The limited progress in diagnostics and treatment greatly impedes the survival of bladder cancer patients. OBJECTIVE: Potential therapeutic biomarkers are urgently needed for future clinical treatment. METHODS: We analyzed the sequencing data and corresponding clinicopathological features and survival information of bladder cancer patients in the TCGA database and identified a new zinc finger protein 485 gene, termed ZNF485, which is highly expressed in the tissues of bladder cancer patients and was verified in cells, animal models and tissue microarrays. RESULTS: We found that inhibition of ZNF485 in the bladder cancer cell lines T24 and 5637 obviously inhibited proliferation and promoted the apoptosis of cancer cells. Furthermore, wound healing and invasion assays showed that downregulation of ZNF485 significantly decreased the mobility and invasion of T24 and 5637 cells. In addition, ZNF485-shRNA transfection obviously inhibited tumor growth in nude mice. Immunohistochemical results of clinical samples showed that the expression level of ZNF485 protein in cancer tissues was higher than that in adjacent tissues. Mechanistic analysis identified possible downstream target genes. CONCLUSIONS: Taken together, the results provide evidence that ZNF485 is involved in bladder cancer proliferation and might be a potential therapeutic biomarker for the treatment of this disease.

LAMTOR5-AS1 regulates chemotherapy-induced oxidative stress by controlling the expression level and transcriptional activity of NRF2 in osteosarcoma cells.

Long-noncoding RNAs (lncRNAs) play roles in regulating cellular functions. High-throughput sequencing analysis identified a new lncRNA, termed LAMTOR5-AS1, the expression of which was much higher in the chemosensitive osteosarcoma (OS) cell line G-292 than in the chemoresistant cell line SJSA-1. Further investigations revealed that LAMTOR5-AS1 significantly inhibits the proliferation and multidrug resistance of OS cells. In vitro assays demonstrated that LAMTOR5-AS1 mediates the interaction between nuclear factor erythroid 2-related factor 2 (NFE2L2, NRF2) and kelch-like ECH-associated protein 1 (KEAP1), which regulate the oxidative stress. Further mechanistic studies revealed that LAMTOR5-AS1 inhibited the ubiquitination degradation pathway of NRF2, resulting in a higher level of NRF2 but a loss of NRF2 transcriptional activity. High level of NRF2 in return upregulated the downstream gene heme oxygenase 1 (HO-1). Moreover, NRF2 controls its own activity by promoting LAMTOR5-AS1 expression, whereas the feedback regulation is weakened in drug-resistant cells due to high antioxidant activity. Overall, we propose that LAMTOR5-AS1 globally regulates chemotherapy-induced cellular oxidative stress by controlling the expression and activity of NRF2.

MiR-34b-3p Represses the Multidrug-Chemoresistance of Bladder Cancer Cells by Regulating the CCND2 and P2RY1 Genes.

BACKGROUND Chemoresistance is a main limitation in chemotherapy for therapeutic cancer. MicroRNA (miRNA) has been indicated in the progression and tumorigenesis of many types of cancer, but the effect of miR-34b-3p in bladder cancer (BCa) cells is still unknown. MATERIAL AND METHODS This research compared the multidrug-sensitive (5637) BCa cell line and the multidrug-resistant (EJ) BCa cell line. We found that CCND2 (G1/S-specific cyclin-D2) and P2RY1 (purinergic receptor P2Y1) were the targets of miR-34b-3p, as further validated by qRT-PCR (quantitative real-time polymerase chain reaction) and western blot analysis. RESULTS Forced reversal of the levels of miR-34b-3p or CCND2/P2RY1 changed the chemoresistance profiles in both 5637 cells and EJ cells. Further experiments suggested that the CCND2 gene and the P2RY1 gene act in concert to negatively correlate with miR-34b-3p effect on BCa multidrug-chemoresistance. CONCLUSIONS These results not only reveal new players regulating BCa chemoresistance, but also provide clues for effective chemotherapy for BCa patients.

Circular RNA SMARCA5 is overexpressed and promotes cell proliferation, migration as well as invasion while inhibits cell apoptosis in bladder cancer.

BACKGROUND: This study aimed to investigate the function of circular RNA SMARCA5 (circ-SMARCA5) on cell proliferation, apoptosis, migration and invasion in bladder cancer. METHODS: Ten pairs of human bladder cancer tissue and adjacent tissue, five human bladder cancer cell lines (including TCCSUP, 5637, J82, UM-UC-3 and T-24) and normal human urothelial cell line SV-HUC-1, were obtained for the detection of circ-SMARCA5. Control overexpression and ShRNA, circ-SMARCA5 overexpression and ShRNA were constructed and transfected into UM-UC-3 cells as Control(+), Control(-), Circ-SMARCA5(+) and Circ-SMARCA5(-) groups. The role of circ-SMARCA5 was investigated in terms of cellular proliferation, apoptosis, migration, and invasion. RESULTS: Circ-SMARCA5 was overexpressed in tumor tissue compared to paired adjacent tissue and it was also overexpressed in TCCSUP, 5637, J82 and UM-UC-3 cells compared to normal human urothelial cell line SV-HUC-1. In UM-UC-3 cells, cell proliferation ability, migration rate and invasion cell count were increased in Circ-SMARCA5(+) group compared to Control(+) group, while reduced in Circ-SMARCA5(-) group compared to Control(-) group. Regarding the cell apoptosis, apoptosis rate and apoptotic protein C-Caspase 3 expression were decreased in Circ-SMARCA5(+) group than those in Control(+) group but raised in Circ-SMARCA5(-) group compared to Control(-) group, meanwhile, the anti-apoptotic protein Bcl-2 expression was elevated in Circ-SMARCA5(+) group than that in Control(+) group but reduced in Circ-SMARCA5(-) group compared to Control(-) group. CONCLUSIONS: Circ-SMARCA5 is overexpressed, and promotes cell proliferation, migration and invasion, but represses apoptosis in bladder cancer.

Extraperitoneal laparoscopic adenomectomy (Madigan) versus bipolar transurethral resection of the prostate for benign prostatic hyperplasia greater than 80 ml: complications and functional outcomes after 3-year follow-up.

PURPOSE: To compare the performance of voluminous benign prostatic hyperplasia patients who have received laparoscopic simple prostatectomy (LSP) with the patients who have received bipolar transurethral resection of the prostate (B-TURP) in their perioperative and 3-year follow-up period. METHODS: Ninety patients with prostate volumes >80 mL (range 80-130 mL) were randomly assigned to either LSP or B-TURP surgery type. The patients were followed up at 1, 3, 6, 12, 24, and 36 months postoperatively. Perioperative and follow-up characteristics were then recorded and compared. RESULTS: More blood loss, greater resected adenoma volume, and shorter catheterization duration were recorded in LSP group than that of B-TURP group (140.1±81.5 vs 93.1±54.0 mL; 65.3±13.8 vs 49.0±12.7 mL; 3.3±1.2 vs 3.8±1.0 days; p<0.05). None of the patients in LSP group reported complications out of 30 days, while 1 case of urethral stricture, 36 cases of retrograde ejaculation, 1 case of bladder neck contracture, and 2 cases of recurrence were recorded in B-TURP group. At 1, 3, 6, and 12 months postoperatively, there were no significant differences in terms of postvoid residual urine volume, maximal urinary flow rate (Qmax), and International Prostate Symptom Score between the two groups (p>0.05). In contrast, the differences became significant at 24 and 36 months (p<0.05). CONCLUSIONS: Compared with B-TURP, LSP with Madigan technique is accompanied by less residual adenoma, shorter catheterization time, and more blood loss. Further, the risk of late complications is lower with LSP and, in terms of functional outcomes, LSP appears to be better than B-TURP beyond 2 years.