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OBJECTIVE: Comparing the laparoscopic pyeloplasty via the mesocolon and para-colonic gutter approach for the treatment of pediatric pelvi-ureteric junction obstruction (UPJO) induced simple hydronephrosis, and analyzing the potential factors influencing surgical outcomes. METHODS: Clinical data of 71 children with UPJO who underwent laparoscopic pyeloplasty at the Department of Urology of the Second Hospital of Hebei Medical University from January 2020 to January 2023 were analyzed. The patients, aged 0.25 to 18 years, were divided into two groups: 30 cases underwent the transcolonic route (mesangial group) and 41 cases underwent the transcolonic paragutter route (paragrow group). RESULTS: showed that both surgical approaches had similar outcomes in terms of operation completion, smooth process, absence of laparotomy, operation time, intraoperative blood loss, postoperative feeding time, and postoperative drainage tube indwelling time, total hospitalization cost, surgical effect, and satisfaction. Common complications such as postoperative fever and abdominal pain were managed with drug treatment or observation, with no need for secondary surgery or fatal complications. Factors such as age, body mass index, preoperative symptoms, severity of hydronephrosis, and ABO blood group classification did not impact the surgical outcome. CONCLUSIONS: There was no statistically significant difference between laparoscopic pyeloplasty and another surgical method in terms of various surgical outcomes for children with ureteropelvic junction obstruction. Factors such as age, body mass index, preoperative symptoms, severity of hydronephrosis, and ABO blood group classification did not have a significant impact on the surgical outcome.
Subject(s)
Hydronephrosis , Laparoscopy , Ureteral Obstruction , Humans , Child , Kidney Pelvis/surgery , ABO Blood-Group System , Laparoscopy/methods , Urologic Surgical Procedures/methods , Ureteral Obstruction/surgery , Ureteral Obstruction/diagnosis , Hydronephrosis/surgery , Treatment Outcome , Retrospective StudiesABSTRACT
OBJECTIVE: The modified day surgery procedure was compared with traditional inpatient procedure and standard day surgery procedure of concealed penile surgery to investigate its advantages, as well as the feasibility of promoting it in our country. METHODS: Retrospective analyzing the clinical data between 135 cases of the concealed penis in children who underwent modified day surgery (day group) and 101 cases who underwent hospitalization surgery (hospitalization group) at the Second Hospital of Hebei Medical University and the results of follow-up.The modified day surgery procedure involves the establishment of dedicated day wards in each surgical department, where the patient's condition is monitored until 8 o'clock the following morning to assess their discharge eligibility.The children's clinical data was divided into two groups to compare clinical parameters, including age at surgery, bleeding volume, operation time, hospitalization expenses, day of hospitalization, and the occurrence of short-term complications before the initial dressing change after surgery.The satisfaction survey of the children was conducted among three distinct groups: the modified day group, the standard day group, and the hospitalization group enabling a comparison of satisfaction levels among these groups. RESULTS: The mean ages of the inpatient and day surgery groups were 8.92±4.42 years old and 11.85±4.43 years old, respectively. No significant differences were observed between these two groups regarding operation time, bleeding volume, and postoperative complications (P>0.05). Compared to the inpatient group, the mean inpatient time and the hospitalization cost of the day group decreased by 69% and 27%, respectively (P<0.05). The patients in the modified procedure group reported the highest satisfaction among the three groups. CONCLUSION: Modified day surgery procedure offers advantages over the standard day surgery procedure and traditional inpatient surgical procedures for the operative treatment of the concealed penis, which makes it suitable for large-scale popularization in China.
Subject(s)
Ambulatory Surgical Procedures , Body Fluids , Child , Male , Humans , Infant, Newborn , Retrospective Studies , China , Device RemovalABSTRACT
OBJECTIVE: To explore the feasibility and safety of cryptorchidism surgery in the day surgery center. METHODS: This retrospective study included 122 cases of unilateral low cryptorchidism (ULC) and 27 cases of bilateral low cryptorchidism (BLC) treated by orchidopexy from July 2018 to July 2022 in the Second Hospital of Hebei Medical University. We divided the patients with ULC into an Ad (day surgery following modified day surgical procedures) and an Ac (conventional surgery) group, and those with BLC into a Bd and a Bc group. We analyzed the clinical data and compared the surgical parameters and patients' satisfaction between different groups. RESULTS: There were no statistically significant differences in the operation age, operation time, intraoperative blood loss, or postoperative complications between the Ad and Ac groups (P > 0.05), but the hospital stay and total cost were markedly reduced in the Ad group by 69% and 10%, respectively, compared with those in the Ac group (P < 0.05). No statistically significant differences were observed between the Bd and Bc groups in the operation age or intraoperative blood loss (P > 0.05), but the Bd group showed significant decreases in the operation time, hospital stay (62%) and total cost (14%) in comparison with the Bc group (P < 0.05). The satisfaction of the patients was remarkably higher in the former than in the latter group. CONCLUSION: Low cryptorchidism surgery following the modified day surgical procedures in the day surgery center is safe and feasible, with the advantages of lower cost and shorter hospital stay.
Subject(s)
Cryptorchidism , Male , Humans , Cryptorchidism/surgery , Retrospective Studies , Ambulatory Surgical Procedures , Feasibility Studies , Blood Loss, Surgical , Treatment OutcomeABSTRACT
BACKGROUND: The dysfunction of myc-related zinc finger protein (MAZ) has been proven to contribute to tumorigenesis and development of multiple cancer types. However, the biological roles and clinical significance of MAZ in clear cell renal carcinoma (ccRCC) remain unclear. METHODS: MAZ expression was examined in ccRCC and normal kidney tissue by quantitative real-time PCR and Western blot. Statistical analysis was used to evaluate the clinical correlation between MAZ expression and clinicopathological characteristics to determine the relationship between MAZ expression and the survival of ccRCC patients. The biological roles of MAZ in cells were investigated in vitro using MTT and colony assays. Luciferase reporter assays and chromatin immunoprecipitation (ChIP) were used to investigate the relationship between MAZ and its potential downstream signaling molecules. RESULTS: MAZ expression is elevated in ccRCC tissues, and higher levels of MAZ were correlated with poor survival of patients with ccRCC. MAZ upregulation elevates the proliferation ability of ccRCC cells in vitro, whereas silencing MAZ represses this ability. Our results further reveal that MAZ promotes cell growth, which is dependent on ERK signaling. Importantly, we found that MAZ positively regulates MAP2K2 expression in ccRCC cells. Mechanistically, MAZ binds to the MAP2K2 promoter and increases MAP2K2 transcription. Furthermore, MAP2K2 levels were shown to be increased in ccRCC tissues and to be associated with a poor prognosis of ccRCC patients. MAP2K2 upregulation activates the ERK signaling pathway and promotes ccRCC progression. CONCLUSION: These results reveal that the MAZ/MAP2K2/ERK signaling axis plays a crucial role in promoting ccRCC progression, which suggests the potential therapeutic utility of MAZ in ccRCC.
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OBJECTIVE: Bladder rupture caused by transurethral clot evacuation is rare in clinic, but an emergency operation is indeed needed in the patient with bladder rupture. We analyzed the reasons of bladder rupture caused by transurethral clot evacuation and provided the countermeasures to guide clinical surgeon to prevent the iatrogenic damage of bladder. METHOD: We retrospectively reviewed the records of 287 patients in our hospital, who had bladder tamponade resulting from clots of blood for various reasons and underwent transurethral clot evacuation from January 2007 to January 2019. Six male cases, aged from 28 to 76 years (mean 56.67±17.76) had bladder rupture. Four patients whose bladder ruptured intraperitoneally were changed to open surgery to repair bladder and clear the remanent blood clots. Two patients with extraperitoneal bladder rupture and a small bladder crevasse underwent a conservative therapy. RESULTS: We observed that the incidence rate of bladder rupture was not associated with bladder tamponade and the age, but may be associated with gender, bladder paracentesis preoperative and urinary retention preoperative. All six cases were male.. They had different period of urinary retention before operation. No supra-pubis bladder paracentesis was made before operation. The bladder crevasses located in the triangle zone and posterior wall of bladder entirely, and the length of the bladder crevasses ranged from 3 to 7cm (mean 4.83cm). The bladder crevasses were all lengthways, and four cases were of' bladders ruptured intraperitoneally while another two presented an extraperitoneal bladder rupture. CONCLUSIONS: The reasons of bladder rupture caused by transurethral clot evacuation may be related to gender, bladder paracentesis preoperative and urinary retention preoperative. We should decide to use expectant treatment or open surgery immediately according to the extent of the rupture when bladder rupture occurs.
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OBJECTIVE: To evaluate the effect and clinical significance of tadalafil combined with atorvastatin on hemodynamics and sexual function in middle-aged and elderly patients with hyperlipidemia complicated with Erectile dysfunction (ED). METHODS: Eighty patients with hyperlipidemia complicated with ED who were treated at The Second Hospital of Hebei Medical University from January 2019 to June 2020 were selected. Using a completely randomized design experimental method, these 80 patients were randomly divided into two groups: the experimental group and the control group, with 40 cases in each group. The control group was treated with a single drug, atorvastatin calcium, while the experimental group was given tadalafil orally on the basis of the control group for 3 months. Changes in the levels of inflammatory factors such as IL-6, TNF and CRP, adverse drug reactions, changes in hemodynamic indicators such as HSV, LSV, PSV, HCT and ESR before and after treatment, as well as changes in sexual function after treatment were compared and analyzed between the two groups. RESULTS: TNF-a, CRP and IL-6 in the experimental group were significantly lower than those in the control group after treatment, with statistically significant differences (p<0.05). There was no significant difference in the incidence of adverse drug reactions between the two groups (p=0.18). After treatment, hemodynamic indexes and sexual function indexes of the experimental group were significantly improved compared with those in the control group, with statistically significant differences (p<0.05). CONCLUSION: A significant improvement effect can be achieved by tadalafil combined with atorvastatin on hemodynamics and sexual function in middle-aged and elderly patients with hyperlipidemia complicated with ED. At the same time, the combination of the two has synergism on inflammatory factors and blood rheology, and the incidence of adverse reactions is not significantly increased.
ABSTRACT
RNA-binding motif protein 5 (RBM5) acts as a tumor suppressor in various human cancers and presents with several important characteristics, such as the potentiation of apoptosis, inhibition of the cell cycle, and alternative splicing of Fas and caspase-2 precursor mRNA. However, its role in bladder urothelial carcinoma (BUC) remains unknown. In this study, we found that RBM5 expression was significantly down-regulated in BUC tissues when compared with the adjacent nontumor tissues. The down-regulation of RBM5 activates ß-catenin, which binds to the T-cell factor/lymphocyte enhancer factor element of the miR-432-5p promoter and elevates the expression of miR-432-5p in bladder cancer cells. The up-regulated miR-432-5p directly targets 3'-UTR and depresses RBM5 expression. Thus, RBM5-miR-432-5p-ß-catenin forms a feedback loop in regulating bladder cancer cell apoptosis. Our findings provide evidence that the regulatory feedback loop among RBM5, miR-432-5p, and Wnt-ß-catenin is responsible for the progress of bladder cancer cells.-Zhang, Y.-P., Liu, K.-L., Wang, Y.-X., Yang, Z., Han, Z.-W., Lu, B.-S., Qi, J.-C., Yin, Y.-W., Teng, Z.-H., Chang, X.-L., Li, J.-D., Xin, H., Li, W. Down-regulated RBM5 inhibits bladder cancer cell apoptosis by initiating an miR-432-5p/ß-catenin feedback loop.
Subject(s)
Apoptosis , Cell Cycle Proteins/genetics , DNA-Binding Proteins/genetics , Feedback, Physiological , MicroRNAs/metabolism , RNA-Binding Proteins/genetics , Tumor Suppressor Proteins/genetics , Urinary Bladder Neoplasms/metabolism , beta Catenin/metabolism , Animals , Cell Cycle Proteins/metabolism , Cell Line, Tumor , DNA-Binding Proteins/metabolism , Down-Regulation , Gene Expression Regulation, Neoplastic , Humans , Male , Mice , Mice, Inbred BALB C , Mice, Nude , MicroRNAs/genetics , RNA-Binding Proteins/metabolism , Tumor Suppressor Proteins/metabolism , Urinary Bladder Neoplasms/genetics , Urothelium/metabolism , beta Catenin/geneticsABSTRACT
OBJECTIVE: To compare the clinical effect of transumbilical single-port laparoscopy combined with improved double hernia needles with that of traditional open surgery in the treatment of hydrocele in children. METHODS: We retrospectively analyzed 35 cases (54 sides) of pediatric hydrocele treated by transumbilical single-port laparoscopy combined with improved double hernia needles (laparoscopy group). We recorded the operation time, intraoperative blood loss, hospital stay, scrotal edema, and postoperative complications and compared them with those of another 46 cases (58 sides) treated by traditional open surgery (open surgery group) during the same period. RESULTS: The laparoscopy group showed a significantly shorter operation time, less intraoperative blood loss, milder scrotal edema, and fewer hospital days than the open surgery group (all P<0.05). However, no statistically significant difference was found in the incidence of postoperative complications between the two groups (P>0.05). Subcutaneous emphysema developed in 2 patients in the laparoscopy group, which disappeared after 1ï¼3 days of oxygen inhalation and other symptomatic treatment, while scrotal hematoma occurred in 1 and incision fat liquefaction in 2 patients in the open surgery group 3 days postoperatively, which healed after debridement suture and daily dressing, respectively. The patients were followed up for 3ï¼6 months, which revealed no late complications in the laparoscopy group but 1 case of unilateral recurrence and 2 cases of offside recurrence in the open surgery group, all cured by laparoscopic internal ring ligation. CONCLUSIONS: Transumbilical single-port laparoscopy combined with improved double hernia needles is superior to traditional open surgery for the treatment of pediatric hydrocele and therefore deserves clinical generalization.
Subject(s)
Laparoscopy/methods , Needles , Testicular Hydrocele/surgery , Blood Loss, Surgical , Child , Edema/diagnosis , Female , Humans , Laparoscopy/instrumentation , Length of Stay , Ligation , Male , Operative Time , Postoperative Complications/diagnosis , Postoperative Complications/surgery , Postoperative Period , Recurrence , Retrospective Studies , Scrotum , Subcutaneous Emphysema/etiology , UmbilicusABSTRACT
Splicing factor 3B subunit 4 (SF3B4) plays important functional roles not only in pre-mRNA splicing, but also in the regulation of transcription, translation, and cell signaling, and its dysregulation contributes to various diseases including Nager syndrome and tumorigenesis. However, the role of SF3B4 and underlying mechanisms in clear cell renal cell carcinoma (ccRCC) remain obscure. In the present study, we found that the expression of SF3B4 was significantly elevated in ccRCC tissues and negatively correlated with the overall survival of ccRCC patients. Upregulation of SF3B4 promotes migration and invasion of ccRCC cells in vitro and in vivo. The promoting effect of SF3B4 on cell migration and invasion is mediated by Twist1, a key transcription factor to mediate EMT. Interestingly, SF3B4, a component of the pre-mRNA spliceosome, is able to promote KLF16 expression by facilitating the transport of KLF16 mRNA into the cytoplasm. Mechanistically, SF3B4 promotes the export of KLF16 mRNA from the nucleus to the cytoplasm and thus enhances KLF16 expression, and in turn elevated KLF16 directly binds to the Twist1 promoter to activate its transcription, leading to EMT and ccRCC progression. Our findings provide evidence that the SF3B4-KLF16-Twist1 axis plays important functional roles in the development and progression of ccRCC, and manipulating this pathway may be a novel therapeutic target for the treatment of ccRCC.
Subject(s)
Carcinoma, Renal Cell , Kidney Neoplasms , Humans , Carcinoma, Renal Cell/metabolism , RNA Precursors/metabolism , RNA, Messenger/genetics , Cytoplasm/metabolism , Cell Line, Tumor , Kidney Neoplasms/metabolism , Cell Proliferation , Gene Expression Regulation, Neoplastic , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Twist-Related Protein 1/genetics , Twist-Related Protein 1/metabolism , Kruppel-Like Transcription Factors/metabolism , RNA Splicing Factors/genetics , RNA Splicing Factors/metabolismABSTRACT
RNA-binding motif protein 24 (RBM24) acts as a multifunctional determinant of cell fate, proliferation, apoptosis, and differentiation during development by regulating premRNA splicing and mRNA stability. It is also implicated in carcinogenesis, but the functions of RBM24 in bladder cancer (BC) remain unclear. In the present study, we revealed that RBM24 was upregulated in BC tissues. Importantly, we found that a higher level of RBM24 was correlated with poor prognosis in BC patients. Overexpression of RBM24 promoted BC cell proliferation, while depletion of RBM24 inhibited BC cell proliferation in vivo and in vitro. Mechanistically, RBM24 positively regulated Runx1t1 expression in BC cells by binding to and enhancing Runx1t1 mRNA stability. Furthermore, Runx1t1 in turn promoted RBM24 expression by interacting with the transcription factor TCF4 and suppressing the transcription of miR-625-5p, which directly targets RBM24 and suppresses RBM24 expression. RBM24-regulated BC cell proliferation was moderated via the Runx1t1/TCF4/miR-625-5p feedback loop. These results indicate that the RBM24/Runx1t1/TCF4/miR-625-5p positive feedback loop participates in BC progression. Disruption of this pathway may be a potential therapeutic strategy for BC treatment.
Subject(s)
Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , MicroRNAs/genetics , RNA-Binding Proteins/genetics , RUNX1 Translocation Partner 1 Protein/genetics , Transcription Factor 4/genetics , Urinary Bladder Neoplasms/genetics , Adult , Aged , Animals , Biomarkers, Tumor , Cell Line, Tumor , Cell Proliferation , Disease Models, Animal , Female , Humans , Male , Mice , Middle Aged , Models, Biological , Neoplasm Grading , Neoplasm Staging , Prognosis , RNA Interference , RNA Stability , RNA-Binding Proteins/metabolism , RUNX1 Translocation Partner 1 Protein/metabolism , Transcription Factor 4/metabolism , Urinary Bladder Neoplasms/metabolism , Urinary Bladder Neoplasms/mortality , Urinary Bladder Neoplasms/pathologyABSTRACT
BACKGROUND: Both E2F transcription factor and cyclin-dependent kinases (CDKs), which increase or decrease E2F activity by phosphorylating E2F or its partner, are involved in the control of cell proliferation, and some circRNAs and miRNAs regulate the expression of E2F and CDKs. However, little is known about whether dysregulation among E2Fs, CDKs, circRNAs and miRNAs occurs in human PCa. METHODS: The expression levels of CDK13 in PCa tissues and different cell lines were determined by quantitative real-time PCR and Western blot analysis. In vitro and in vivo assays were preformed to explore the biological effects of CDK13 in PCa cells. Co-immunoprecipitation anlysis coupled with mass spectrometry was used to identify E2F5 interaction with CDK13. A CRISPR-Cas9 complex was used to activate endogenous CDK13 and circCDK13 expression. Furthermore, the mechanism of circCDK13 was investigated by using loss-of-function and gain-of-function assays in vitro and in vivo. RESULTS: Here we show that CDK13 is significantly upregulated in human PCa tissues. CDK13 depletion and overexpression in PCa cells decrease and increase, respectively, cell proliferation, and the pro-proliferation effect of CDK13 is strengthened by its interaction with E2F5. Mechanistically, transcriptional activation of endogenous CDK13, but not the forced expression of CDK13 by its expression vector, remarkably promotes E2F5 protein expression by facilitating circCDK13 formation. Further, the upregulation of E2F5 enhances CDK13 transcription and promotes circCDK13 biogenesis, which in turn sponges miR-212-5p/449a and thus relieves their repression of the E2F5 expression, subsequently leading to the upregulation of E2F5 expression and PCa cell proliferation. CONCLUSIONS: These findings suggest that CDK13 upregulation-induced formation of the positive feedback loop among circCDK13, miR-212-5p/miR-449a and E2F5 is responsible for PCa development. Targeting this newly identified regulatory axis may provide therapeutic benefit against PCa progression and drug resistance.
Subject(s)
CDC2 Protein Kinase/metabolism , E2F5 Transcription Factor/metabolism , MicroRNAs/metabolism , Prostatic Neoplasms/metabolism , CDC2 Protein Kinase/genetics , Cell Proliferation/physiology , E2F5 Transcription Factor/genetics , Feedback , Female , Humans , Male , MicroRNAs/genetics , Prostatic Neoplasms/genetics , Prostatic Neoplasms/pathology , Transfection , Up-RegulationABSTRACT
Accumulating evidence has demonstrated that thioredoxin interacting protein (TXNIP) is abnormally expressed in a variety of malignant tumors and functions as a tumor suppressor. However, the association between TXNIP and clear cell renal cell carcinoma (CCRCC) has not yet been fully elucidated. The aim of the present study was to evaluate the role of TXNIP in CCRCC using The Cancer Genome Atlas (TCGA) database. The RNA sequencing data and corresponding clinical data were collected from TCGA database. The association between TXNIP and patient clinicopathological characteristics was analyzed using analysis of variance and logistic regression. The Kaplan-Meier method and Cox proportional hazards model were used to assess the association between TXNIP and overall survival. Gene Set Enrichment Analysis (GSEA) was used to explore the associated signaling pathways. TXNIP expression was identified to be decreased in CCRCC tissues compared with normal tissues. The decreased expression of TXNIP in CCRCC was significantly associated with clinical stage [OR=0.509 for III vs. I (P=0.002); OR=0.527 for IV vs. I (P=0.012)], T stage [OR=0.552 for T3 vs. T1 (P=0.002)] and grade [OR=0.261 for G4 vs. G1 (P=0.027)]. Kaplan-Meier survival analysis indicated that cases of CCRCC with low TXNIP expression were associated with poorer prognoses compared with those with a high expression level (P=0.002). Univariate and multivariate Cox analyses indicated that TXNIP was an independent prognostic factor in CCRCC. GSEA revealed that 6 pathways exhibited significant differential enrichment in the TXNIP high-expression phenotype, including the WNT signaling pathway, the mitogen-activated protein kinase (MAPK) signaling pathway, the phosphatidylinositol signaling system, the transforming growth factor-ß (TGF-ß) signaling pathway, autophagy and the Janus kinase (JAK)-STAT signaling pathway. Taken together, the results of the present study indicate that TXNIP expression may be a potential prognostic marker for patients with CCRCC. In addition, the WNT signaling pathway, MAPK signaling pathway, phosphatidylinositol signaling system, TGF-ß signaling pathway, autophagy and the JAK-STAT signaling pathway may be the crucial pathways regulated by TXNIP in CCRCC.
ABSTRACT
Nicotinic acetylcholine receptor (nAChR) subunit α5 (α5nAChR) is involved in tumor cell proliferation, inhibition of apoptosis, progression of metastasis, and induction of angiogenesis in certain solid tumors. However, the role of α5nAChR in prostate cancer cell growth and metastasis is unclear. In the present study, the role of α5nAChR in cell proliferation, migration, invasion and apoptosis was investigated by silencing the expression levels of α5nAChR in the prostate cancer cell lines DU145 and PC3. A siRNA oligonucleotide targeting α5nAChR was designed. The cell proliferation of DU145 and PC3 cell lines was analyzed by the Cell Counting Kit8 (CCK8) assay. Cell migratory and invasive activities were determined using wound healing and Transwell assays, respectively. Western blot analysis was used to quantify α5nAChR, pAKT and pERK1/2 levels in DU145 and PC3 cells. Knockdown of α5nAChR was associated with decreased cell proliferation, migration, invasion and increased apoptosis. In addition, decreased phosphorylation levels of AKT and ERK1/2 were revealed following α5nAChR knockdown in DU145 and PC3 cells compared with those observed in the scramble control samples. The expression levels of the apoptosisrelated proteins were altered following silencing of α5nAChR. In summary, the data indicated that α5nAChR was involved in the proliferation and invasion of human prostate cancer cells.
Subject(s)
Prostatic Neoplasms/pathology , Receptors, Nicotinic/genetics , Receptors, Nicotinic/metabolism , Up-Regulation , Aged , Biomarkers, Tumor/genetics , Biomarkers, Tumor/metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , Gene Silencing , Humans , Male , Middle Aged , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Neoplasm Invasiveness , PC-3 Cells , Phosphorylation , Prognosis , Prostatic Neoplasms/genetics , Prostatic Neoplasms/metabolismABSTRACT
The transcription factor E2F1 regulates the expression of the miR-20b-5p precursor and is involved in epithelial-to-mesenchymal transition (EMT). Transforming growth factor-ß1 (TGF-ß1) induces EMT in prostate cancer (PCa) by binding to TGF-beta receptor 2 (TGFBR2) to activate TGF-ß signaling. However, the relationship between TGFBR2, E2F1, and miR-20b-5p in the modulation of EMT in PCa cells remains unknown. In this study, we found that the level of miR-20b-5p expression was significantly lower in PC3 and DU145 cells than that in prostate epithelial (RWPE-1) cells, and TGF-ß1 treatment further down-regulated miR-20b-5p expression in these two cell lines. Functional studies showed that miR-20b-5p suppressed TGF-ß1-induced migration and invasion of PC3 and DU145 cells by up-regulating E-cadherin and down-regulating vimentin, leading to TGF-ß1-induced inhibition of EMT. Using gain and loss of function experiments, it was shown that E2F1 mediated TGF-ß1 regulation of miR-20b-5p expression. Further, a luciferase activity assay showed that TGFBR2 was a direct target of miR-20b-5p in PCa cells. These results suggest that miR-20b-5p, TGFBR2, and E2F1 form a regulatory loop to modulate EMT induced by TGF-ß1. A novel regulatory mechanism underlying the miR-20b-5p/TGFBR2/E2F1 axis is involved in TGF-ß1-induced EMT of PCa cells, and miR-20b-5p may be a potential therapeutic target for PCa.
ABSTRACT
Prostate cancer (PCa) is a frequently occurring malignancy in males, and epithelial mesenchymal transition (EMT) plays a critical role in PCa metastasis. Thus, developing biomarkers inhibiting EMT may provide significance for treatment of PCa. Hence, the aim of the current study was to investigate the mechanism by which FBP1 gene silencing influences PCa cell EMT, invasion and metastasis by mediating the MAPK pathway. PCa cell lines exhibiting the highest FBP1 expression were selected and treated with plasmids of siRNA-FBP1 sequence 1 and 2, pcDNA3.1-Flag-FBP1 (over-expression plasmid of FBP1), U0126 (an inhibitor of the ERK signaling pathway) and PD98059 (an inhibitor of the MEK signaling pathway). Cell proliferation, migration and invasion were detected by MTT assay, wound healing assay and Transwell assay, respectively. The mRNA and protein expression of related factors of EMT and MAPK signaling were determined by RT-qPCR and western blot analysis, respectively. Xenograft tumor growth after inoculation of DU145 cells was regularly analyzed in the nude mice. The positive expression of EMT markers was determined by immunohistochemistry. DU-145 and PC-3 cells displaying the highest FBP1 expression were selected for further analysis. The PCa cells treated with siRNA-FBP1 exhibited increased proliferation, migration rate and invasion, in addition to facilitated xenograft tumor growth. Notably, siRNA-FBP1 was identified to accelerate PCa cell EMT by elevating the expression of Vimentin and N-cadherin while diminishing E-cadherin expression via activation of the MAPK signaling pathway. The aforementioned results were reversed in PCa cells treated by pcDNA3.1-Flag-FBP1. Evidence has been provided in this study that FBP1 gene silencing activates the MAPK pathway, which ultimately promotes cell EMT, invasion and metastasis in PCa.
Subject(s)
Epithelial-Mesenchymal Transition/genetics , MAP Kinase Signaling System/genetics , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/pathology , Animals , Cadherins/genetics , Cadherins/metabolism , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Fructose-Bisphosphatase/genetics , Fructose-Bisphosphatase/metabolism , Gene Expression Regulation, Neoplastic/genetics , Gene Knockdown Techniques , Gene Silencing , Humans , MAP Kinase Kinase Kinases/antagonists & inhibitors , MAP Kinase Kinase Kinases/metabolism , MAP Kinase Signaling System/drug effects , Male , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness/genetics , Neoplasm Metastasis , Phosphorylation , Prostatic Neoplasms/enzymology , Prostatic Neoplasms/genetics , RNA, Small Interfering , Transplantation, Heterologous , Vimentin/genetics , Vimentin/metabolismABSTRACT
OBJECTIVE: To analyze the invention effect of curcumin on chronic urinary tract infection in rats and explore its possible mechanism of action. METHODS: The experimental animals were randomly divided into three groups, normal, model and curcumin group. Chronic urinary tract infection models were built for model group and curcumin group by injecting coliform fluid into the cavity of bladder. From the first day of modeling, rats in the curcumin group were injected with 150 mg/kg curcumin, while rats in normal group and model group were given no other treatment. The treatment lasted for 14 d. The white blood cell counts in blood and urine, bacterial colony count in urine and renal tubular functional indexes of rats in all groups at day 1, 7, and 14 after treatment were detected. Urine ß2-microglobulin (ß2-MG), urinary N-acetyl-ß-D glucosaminidase (NAG) levels were used to detected the inflammatory cytokines in serum after treatment including the contents of IL-6, IL-8, IL-10 and monocyte chemoattractant protein-1 (MCP-1), and real-time PCR was employed to determine the expression of mRNA of toll-like receptor 2 (TLR-2) and TLR-4 in renal tissues and bladder tissues of all groups after treatment. RESULTS: The white blood cell counts at day 1 and 7 after treatment in rats of model group and curcumin group were significantly higher than those of normal group at the same time points, while the white blood cell counts of the curcumin group were significantly lower than those of model group (P < 0.05). The urine white blood cell counts in rats of model group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points; those in the curcumin group were significantly lower than those of the model group at day 1, 7 and 14 at the same time points (P < 0.05). The bacterial colony counts of urine in rats of model group and curcumin group at day 1, 7 and 14 were all significantly higher than those of normal group at the same time points, while the counts of curcumin group were significantly lower than those of model group at the same time points (P < 0.05). Levels of urine ß2-MG, NAG, IL-6, IL-8, IL-10, MCP-1 and expression of TLR2 mRNA and TLR4 mRNA in renal and bladder tissues in rats of model group were significantly higher than those of the normal group, while these variables of the curcumin group were significantly higher than those of the normal group but lower than those of model group (P < 0.05). CONCLUSIONS: Curcumin can significantly improve the symptoms of chronic urinary tract infections, protect renal tubular function, and also decline inflammatory responses by influencing the expressions of TLR2 mRNA and TLR4 mRNA so as to exert its curative effect on chronic urinary tract infections.
ABSTRACT
OBJECTIVE: To approach the treatment efficiency of replication defective adenovirus carrying HSV-tk gene under control of the human telomerase reverse transcriptase (hTERT) promoter in a mouse xenografts model of prostate cancer. METHODS: It was erected that the model of human prostate cancer in BALB/C nude mouse followed by locally injecting Ad-hTERT-HSV-tk and peritoneal injection of GCV. The anti-tumor efficacy was evaluated by index of tumor volume, tumor weight, relative tumor curative, and tumor growth curve. Afterwards, the TUNEL and transmission electron microscope to overview apoptosis of tumor cells were used. Finally, immunohistochemistry for detecting PCNA of tumor cells were applied. RESULTS: Compared with the control group, all viruses treatment groups demonstrated tumor growth inhibition. Among 3 treatment groups, antitumoral effect of Ad-hTERT-HSV-tk/GCV was much stronger than other two groups (P < 0.05). Obvious necrosis and apoptosis was observed by TUNEL, and PCNA was detected by immunohistochemistry in tumor tissues in all viruses treatment group. CONCLUSIONS: Ad-hTERT-HSV-tk/GCV system is highly efficacious to inhabit the growth of human prostate cancer.