LIMD2 promotes tumor proliferation, invasion, and epithelial-mesenchymal transition in clear cell renal cell carcinoma.

LIMD2 was found upregulated in various tumors and metastatic samples and associated with a poor prognosis. But the role of LIMD2 in clear cell renal cell carcinoma (ccRCC) remains elusive. The expression of LIMD2 in ccRCC was analyzed using cohort data downloaded from TCGA and ICGC databases. In vitro and in vivo experiments were then conducted to study the biological role of LIMD2 in ccRCC and explore the possible mechanism. The results indicated that LIMD2 was overexpressed and correlated with a poor outcome in ccRCC. LIMD2 promoted the malignancy of ccRCC both in vitro and in vivo. LIMD2 induced epithelial-mesenchymal transition (EMT) via activating the ILK/Akt pathway in ccRCC. In conclusion, LIMD2 is overexpressed and promotes proliferation, invasion, and EMT in ccRCC, which may serve as a potential novel therapeutic target for ccRCC.

CircPVT1 promotes progression in clear cell renal cell carcinoma by sponging miR-145-5p and regulating TBX15 expression.

Emerging evidence revealed that circular RNAs (circRNAs) play significant roles in regulating tumorigenesis and cancer progression. However, few circRNAs were well characterized in clear cell renal cell carcinoma (ccRCC). We found that circPVT1 was significantly upregulated in ccRCC tissues and positively associated with the clinical stage. The Area Under Curve of tissue and serum circPVT1 expression in ccRCC were 0.93 and 0.86, respectively. Importantly, we demonstrated that circPVT1 promoted ccRCC growth and metastasis in vitro and in vivo. We also found that circPVT1 directly binds to miRNA-145-5p via the Biotin-labelled miRNA pulldown assay and dual-luciferase reporter assay, and miR-145-5p inhibitor significantly attenuated the effect of circPVT1 knockdown on ccRCC cells. Moreover, through RNA sequencing and bioinformatics analysis, we demonstrated that TBX15 was regulated by the circPVT1/miR-145-5p axis and predicted poor prognosis in ccRCC. These findings suggest that circPVT1 promotes ccRCC growth and metastasis through sponging miR-145-5p and regulating downstream target TBX15 expression. The circPVT1/miR-145-5p/TBX15 axis might be a potential diagnostic marker and therapeutic target in ccRCC.

M2 macrophages secrete CXCL13 to promote renal cell carcinoma migration, invasion, and EMT.

OBJECTIVE: M2 macrophages are associated with a poor prognosis in a variety of malignancies. There are, however, few relevant investigations in clear cell renal cell carcinoma (ccRCC). METHODS: The expression of M2 macrophages in ccRCC tissues was first discovered using immunohistochemistry in this study. Then, M2 macrophages were created in vitro to see how they affected the proliferation, migration, invasion, and EMT of ccRCC cells. Using qPCR and prognostic analysis identifies important chemokine. Antibody neutralization tests confirmed the chemokine's involvement and function. Pathway inhibitors confirmed the main pathway of M2 macrophages in ccRCC. Finally, qPCR and IHC were used to confirm the expression of chemokine receptors in ccRCC tissues. RESULTS: The presence of M2 macrophages was linked to a poor outcome in ccRCC. M2 macrophages enhanced the proliferation, migration, invasion, and EMT of ccRCC lines in vitro. CXCL13 was identified as the main chemokine by prognostic analysis and qPCR tests. CXCL13 neutralizing antibodies can inhibit the stimulation of M2 macrophages in ccRCC lines' proliferation, migration, invasion, and EMT. M2 macrophages and CXCL13 may activate the Akt pathway in ccRCC lines, and Akt inhibitors decrease ccRCC lines proliferation, migration, invasion, and EMT. CXCR5 expression is a poor prognostic factor for renal cell carcinoma, according to qPCR and immunohistochemistry. In vivo experiments further proved that CXCL13 secreted by M2 macrophages can promote tumor proliferation. CONCLUSIONS: M2 macrophages in the immunological milieu secrete CXCL13, which promotes ccRCC proliferation, migration, invasion, and EMT. Our findings contribute to a better understanding of the function of the tumor microenvironment in the incidence and progression of ccRCC, and they may point to novel therapeutic targets for ccRCC.

Development and validation of a CT-based nomogram for preoperative prediction of clear cell renal cell carcinoma grades.

OBJECTIVES: Nuclear grades are proved to be one of the most significant prognostic factors for clear cell renal cell carcinoma (ccRCC). Radiomics nomogram is a widely used noninvasive tool that could predict tumor phenotypes. In this study, we performed radiomics analysis to develop and validate a CT-based nomogram for the preoperative prediction of nuclear grades in ccRCC. METHOD: CT images and clinical data of 258 ccRCC patients were retrieved from the Cancer Imaging Archive (TCIA). Radiomics features were extracted from arterial-phase CT images using 3D Slicer software. LASSO regression model was performed to develop a radiomics signature in the training set (n = 143). A radiomics nomogram was constructed combining radiomics signature and selected clinical predictors. Receiver operating characteristic (ROC) curve and calibration curve were used to determine the performance of the radiomics nomogram in the training and validation set (n = 115). Decision curve analysis was used to assess the clinical usefulness of the CT-based nomogram. RESULTS: One thousand three hundred sixteen radiomics features were extracted from arterial-phase CT images. A radiomics signature, consisting of 20 features, was developed and showed a favorable performance in discriminating nuclear grades with an area under the curve (AUC) of 0.914 and 0.846 in the training and validation set, respectively. The CT-based nomogram, including the radiomics signature and the CT-determined T stage, achieved good calibration and discrimination in the training set (AUC, 0.929; 95% CI, 0.886-0.972) and validation set (AUC, 0.876; 95% CI, 0.812-0.939). Decision curve analysis demonstrated the clinical usefulness of the CT-based nomogram. CONCLUSION: The noninvasive CT-based nomogram, including radiomics signature and CT-determined T stage, could improve the accuracy of preoperative grading of ccRCC and provide individualized treatment for ccRCC patients. KEY POINTS: • Contrast-enhanced CT may help in preoperative grading of ccRCC. • The CT-based nomogram incorporated a radiomics signature and CT-determined T stage could preoperatively predict ccRCC grades. • The CT-based nomogram has the potential to improve individualized treatment and assist clinical decision making of ccRCC patients.

A nomogram to predict stricture-free survival in patients with ureteral stricture after balloon dilation.

BACKGROUND: Balloon dilation is a commonly used minimally invasive endourological treatment of ureteral stricture, but the postoperative recurrence rate is relatively high. And factors contributing to recurrence after treatment are poorly understood. Herein, we sought to develop a novel clinical nomogram to predict ureteral stricture-free survival in patients suffering from ureter stricture and performed balloon dilation. METHODS: The nomogram was established based on a retrospective analysis of 321 patients who received endoscopic balloon dilation alone for ureter strictures from January 2016 to January 2020 in Sun Yat-sen Memorial Hospital using the Cox regression model. Perioperative clinical data and disease outcomes were analyzed. The primary endpoint was the onset of ureteral re-stricture after ureter balloon dilation. Discrimination of the nomogram was assessed by the concordance index (C-index) and the calibration curve. The results were internally validated using bootstrap resampling. RESULTS: Overall, 321 patients with a median follow-up of 590 days were enrolled in the study, among which 97 patients (30.2%) developed recurrence of ureteral stricture during follow-up. Five variables remained significant predictors of ureteral re-stricture after multivariable analyses: stricture nature (P < 0.001), urinary nitrite (P = 0.041), CKD (P = 0.005), stent retention time (P < 0.001), and balloon size (P = 0.029). The calibration craves for the probability of 1-, 3-, and 5-years stricture-free survival (SFS) presented satisfied with the consistency of nomogram prediction and actual observation. The C-index of the model was 0.74 (95% CI 0.70-0.79). CONCLUSIONS: Our study developed the first nomogram to effectively predict stricture-free survival in patients suffering from ureter stricture after balloon dilation. It is helpful to identify the optimal patients with ureter stricture for balloon dilation and improve treatment outcomes. However, further external validation of the nomogram is warranted.

Long Non-Coding RNA LUCAT1 Promotes Proliferation and Invasion in Clear Cell Renal Cell Carcinoma Through AKT/GSK-3ß Signaling Pathway.

BACKGROUND/AIMS: Long non-coding RNAs (lncRNAs) have emerged as new regulators and biomarkers in several cancers. However, few lncRNAs have been well characterized in clear cell renal cell carcinoma (ccRCC). METHODS: We investigated the lncRNA expression profile by microarray analysis in 5 corresponding ccRCC tissues and adjacent normal tissues. Lung cancer-associated transcript 1 (LUCAT1) expression was examined in 90 paired ccRCC tissues by real-time PCR and validated by The Cancer Genome Atlas (TCGA) database. Kaplan-Meier analysis was used to examine the prognostic value of LUCAT1 and CXCL2 in ccRCC patients. Loss and gain of function were performed to explore the effect of LUCAT1 on proliferation and invasion in ccRCC cells. Western blotting was performed to evaluate the underlying mechanisms of LUCAT1 in ccRCC progression. Chemokine stimulation assay was performed to investigate possible mechanisms controlling LUCAT1 expression in ccRCC cells. Enzyme-linked immunosorbent assays were performed to determine serum CXCL2 in ccRCC patients and healthy volunteers. Receiver operating characteristic curve analysis was performed to examine the clinical diagnostic value of serum CXCL2 in ccRCC. RESULTS: We found that LUCAT1 was significantly upregulated in both clinical ccRCC tissues (n = 90) and TCGA ccRCC tissues (n = 448) compared with normal tissues. Statistical analysis revealed that the LUCAT1 expression level positively correlated with tumor T stage (P < 0.01), M stage (P < 0.01), and TNM stage (P < 0.01). Overall survival and disease-free survival time were significantly shorter in the high-LUCAT1-expression group than in the low-LUCAT1-expression group (log-rank P < 0.01). LUCAT1 knockdown inhibited ccRCC cell proliferation and colony formation, induced cell cycle arrest at G1 phase, and inhibited cell migration and invasion. Overexpression of LUCAT1 promoted proliferation, migration, and invasion of ccRCC cells. Mechanistic investigations showed that LUCAT1 induced cell cycle G1 arrest by regulating the expression of cyclin D1, cyclin-dependent kinase 4, and phosphorylated retinoblastoma transcriptional corepressor 1. Moreover, LUCAT1 promoted proliferation and invasion in ccRCC cells partly through inducing the phosphorylation of AKT and suppressing the phosphorylation of GSK-3ß. We also revealed that chemokine CXCL2, upregulated in ccRCC, induced LUCAT1 expression and might be a diagnostic and prognostic biomarker in ccRCC. CONCLUSIONS: LUCAT1 was upregulated in ccRCC tissues and renal cancer cell lines, and significantly correlated with malignant stage and poor prognosis in ccRCC. LUCAT1 promoted proliferation and invasion in ccRCC cells through the AKT/GSK-3ß signaling pathway. We also revealed that LUCAT1 overexpression was induced by chemokine CXCL2. These findings indicate that the CXCL2/LUCAT1/AKT/GSK-3ß axis is a potential therapeutic target and molecular biomarker for ccRCC.

Laparoscopic Partial Nephrectomy for T1 Renal Cell Carcinoma: Comparison of Two Resection Techniques in a Multi-institutional Propensity Score-Matching Analysis.

BACKGROUND: Enucleoresection is defined as presence of a minimal paratumor parenchyma that allows for clear visualization of the tumor's contours during partial nephrectomy (PN). Because there is variability in published reports regarding resection techniques during PN before the surface-intermediate-base (SIB) margin score reporting system, the association between postoperative outcomes and resection techniques are rarely reported. This study was designed to compare the perioperative, oncologic, and functional outcomes between laparoscopic enucleoresection (LER) (SIB score 1 + 1 + 1 = 3) and traditional laparoscopic partial nephrectomy (TLPN) (SIB score 1 + 2 + 2 = 5). METHODS: Data from 270 consecutive patients who underwent laparoscopic partial nephrectomy for single T1 RCC at 3 medical centers were prospectively collected. Propensity score matching was performed on age, gender, body mass index, American Society of Anesthesiologists score, preoperative estimated glomerular filtration rate (eGFR), tumor size, RENAL nephrometry score, Charlson score, and solitary kidney status. Normal parenchyma width of each patient was evaluated right after the surgery, and SIB score was assigned retrospectively. Ninety-eight matched patients undergoing LER or TLPN were compared for perioperative, oncologic, and functional outcomes. RESULTS: After matching, warm ischemia time (WIT) and operative time were significantly shorter in LER than TLPN group (20.8 vs. 23.8 min, P = 0.003 and 130.8 vs. 152.1 min, P = 0.005, respectively). Estimated blood loss (EBL) also was lower in LER than TLPN group (50 vs. 90 mL, P = 0.045). Complication rates, positive surgical margin rates, and local recurrence rates were comparable between groups (P = 0.3, P = 0.62, and P = 1.0, respectively). At last follow-up, the eGFRs also were comparable in both groups (P = 0.6). CONCLUSIONS: LER has similar oncologic, functional outcomes and complication rates with the advantage of a shorter WIT, operative time, and lower EBL compared with TLPN.

Angiokeratoma of Fordyce response to long pulsed Nd:YAG laser treatment.

Angiokeratoma of Fordyce can be easily diagnosed by their typical erythra and cured by varied therapeutic methods including surgery, electrocoagulation, cryotherapy, or various laser systems. Which are the optimal therapeutic methods? There are no consentaneous opinions in the reported articles. We present our characteristic treatment with the application of long pulsed neodymium-dopedyttrium-aluminum-garne (lpNd:YAG) laser in the treatment of angiokeratoma of Fordyce. A 1064 nm lpNd:YAG laser (spot size: 5 mm/7 mm, energy:90-130 J/cm(2) , and pulse duration: 10-20 ms.) was used to treat the patient's lesions. The desirable clinical endpoint of the treatment was lesions shrunk and turned pallor immediately after the irradiation. The treatment interval was at least 8 weeks. Of the 11 patients, 9 of them were cured and 2 of them were improved. The mean treatment sessions were 2.2 times. None of them had a scar formation and any other side effects. All of them were satisfied with the treatment results. We conclude that angiokeratoma of Fordyce responded well to lpNd:YAG laser treatment. It provided a simple, rapid, and no bleeding treatment in treating Angiokeratoma of Fordyce.

Extraperitoneal transumbilical laparoendoscopic single-site radical prostatectomy using a homemade single-port device: 20 cases with midterm outcomes.

PURPOSE: To describe the surgical technique and to report the midterm outcomes of laparoendoscopic single-site radical prostatectomy (LESS-RP) with a homemade single-port device. METHODS: Between August 2009 and October 2011, 20 LESS-RP procedures were performed in a single center by a high-volume surgeon. The surgical methods are outlined in the paper. Preoperative, perioperative, pathologic, midterm oncological, and functional outcomes data were analyzed. RESULTS: On average, patients were 66.1 years old, had a mean body mass index of 24.4 kg/m(2), and a mean prostate-specific antigen (PSA) value of 12.2 ng/ml. No conversion was required; however, a single additional 5-mm port was required in one case. The mean operative time was 248 min; mean estimated blood loss was 94 ml. The mean visual analog pain score at day 3 was 1.5 out of 10, and the mean hospital stay was 15.4 day. Pathology showed one focal positive margin (5 %) in a T3a patient, and mean Gleason score was 6.7. There were a total of four Clavien grade 2 complications. The median follow-up was 22.8 month (range 12-36 month). No PSA recurrence was detected during follow-up period; the potency rate after bilateral neurovascular bundle preservation was 60 % (3/5). Complete continence recovery (no pad) was observed in 85 % of the patients at 1 year, in 90 % at 2 years, and in all three patients at 3 years. CONCLUSIONS: LESS-RP is technically feasible and safe, with excellent cosmesis, low postoperative pain levels, and acceptable midterm oncological, and functional outcomes. Comparative investigation to conventional LRP is needed.

Knockdown BMI1 expression inhibits proliferation and invasion in human bladder cancer T24 cells.

B cell-specific moloney murine leukemia virus integration site 1 (BMI1) is a transcriptional repressor of polycomb repressive complex 1, which is involved in the proliferation, senescence, migration, and tumorigenesis of cancer. Experimental researchers have convincingly linked BMI1 to tumorigenesis. However, there is no study about the issue on the role of BMI1 in the proliferation, apoptosis, and migration of bladder cancer. To address this question, we examined the expression of BMI1 in bladder cancer tissues and used siRNA to knockdown BMI1 expression in bladder cancer T24 cells. Then we tested the cell proliferation by CCK8 assay and soft agar colony formation assay, apoptosis by flow cytometry assay, and cell invasiveness by transwell migration assay. Our results revealed that BMI1 promoted proliferation, migration, invasion, and progression in bladder cancer. Over-expression of BMI1 was correlated with tumor clinic-pathological features. BMI1 siRNA effectively inhibited bladder cancer cell proliferation and migration in vitro, and it promoted bladder cancer invasion, maybe by causing epithelial-to-mesenchymal transition. Our findings suggested that BMI1 may represent a novel diagnostic marker and a therapeutic target for bladder cancer, and deserves further investigation.

Contemporary Research Progress on the Detection of Polycyclic Aromatic Hydrocarbons.

Polycyclic aromatic hydrocarbons (PAHs) are a class of the most common and widespread contaminants. The accumulation of PAHs has made a certain impact on the environment and is seriously threatening human health. Numerous general analytical methods suitable for PAHs were developed. With the development of economy, the environmental problems of PAHs in modern society are more extensive and prominent, and attract more attention from environmental scientists and analysts. Deeper understanding of the properties of PAHs depends on the advent of detection methods, which can also be more conducive to promoting the protection of the environment. Till now, more sensitive, more high-speed and more high-throughput analytical tools are being invented and have played important roles in the research of PAHs. In this short review article, we focused mainly on the contemporary analytical methods about PAHs. We started with a brief review on the hazards, migration, distribution and traditional analysis methods of PAHs in recent years, including liquid chromatography, gas chromatography, surface enhanced Raman spectroscopy and so on. We also presented the applications of the modern ambient mass spectrometry, especially microwave plasma torch mass spectrometry, in the detection of PAHs, as well as the far out novel results in our lab by using microwave plasma torch (MPT) mass spectrometry; for example, some new insights about Birch reduction, regular hydrogen addition and the robustness of molecular structure. These studies have demonstrated the versatility of MPT MS as a platform in the research of PAHs.

Conditional survival of trimodal therapy for nonmetastatic muscle-invasive bladder cancer: A SEER database analysis.

OBJECTIVE: Conventional survival analysis plays a limited role in patients who have survived a period after initial treatment. The present study analyzed how conditional survival (CS) predicted survival rate over time for nonmetastatic muscle-invasive bladder cancer (MIBC) patients after trimodal treatment. METHOD: This retrospective study from the SEER database included consecutive patients with nonmetastatic MIBC who received trimodal therapy (TMT) between January 2010 and December 2017. Kaplan-Meier analysis was used to estimate overall survival (OS) and cancer-specific survival (CSS). CS was defined as the rate of surviving y years after already surviving for x years. Multivariate Cox regression analysis was used to identify prognostic factors. RESULT: A total of 1110 nonmetastatic MIBC patients treated with TMT were included. Given a 1-, 2-, 3-, and 4-year after TMT, the rate of surviving to 5-year, respectively, improved by +5.0 (20.0%), +17.0 (32.0%), +30.0 (45.0%), and +52.8 (67.8%) from those calculated at baseline (15.0%). The 2-year CS rate of patients who had survived 1-, 2-, or 3-year after TMT improved, respectively, compared to 3-, 4-, or 5-year actual survival. Multivariate Cox regression analysis demonstrated that adverse variables (T stage, age) of OS and CSS lost their prognostic significance over time. DISCUSSION AND CONCLUSION: Conditional survival rate of surviving to 5-year after TMT kept a relatively stable level over time. In addition, those adverse variables were not always the prognostic factors over time. Only age was always the significant prognostic factor for conditional OS from baseline to 5-year survival. Our results provided real-time survival information and prognosis estimates to adjust follow-up plans for nonmetastatic MIBC patients after TMT.

Selection of Optimal Candidates for Cytoreductive Nephrectomy in Patients with Metastatic Clear Cell Renal Cell Carcinoma: A Predictive Model Based on SEER Database.

BACKGROUND: Currently, the progress of targeted drugs in the treatment of metastatic clear cell renal cell carcinoma (mccRCC) is limited. Cytoreductive nephrectomy (CN), as an alternative treatment, can improve the prognosis of patients with metastatic renal cell carcinoma to some extent. However, it is unclear which patients would benefit from this tumor reduction operation. As a consequence, we developed a predictive model to identify patients who may well benefit from CN in terms of survival. METHODS: We identified patients with metastatic clear cell renal cell carcinoma retrospectively from the Surveillance, Epidemiology, and End Results (SEER) database (2010-2015) and classified them into surgery and non-surgery groups. Propensity score matching (PSM) was performed to balance the baseline characteristics. Patients who survived longer than the median overall survival (OS) of no-surgery group were defined as surgical-benefit patients. Then, we developed a predictive model based on preoperative characteristics using multivariable Logistic regression. Calibration curves and the area under the receiver operating characteristic (AUC) were used to evaluate the efficiency of the predictive model. The clinical value of the nomogram was assessed utilizing decision curve analysis (DCA). RESULTS: Our study collected 5544 patients from the SEER database, with 2352(42.4%) receiving cytoreductive surgery. Overall survival (OS) was longer in the CN group than in the non-surgery group after 1:1 propensity scoring matching (median OS: 19 months vs 7 months; hazard ratio (HR) =0.4106, P< 0.001). In the matched surgery group, 65.7% (367) patients survived more than 7 months after the operation and they were considered to benefit from CN. The predictive model performed well on both the training group (AUC=73.4%) and the validation group (AUC=71.9%) and the calibration curves indicated a high degree of consistency. The decision curve analysis curve demonstrated the clinical utility. We classified surgical patients into the beneficial group and non-beneficial group by using the predictive model, then discovered a substantial difference in OS between the two groups. CONCLUSIONS: We developed a nomogram to select ideal mccRCC patients who might benefit from cytoreductive nephrectomy. Clinicians could make a more precise treatment strategy for mccRCC patients.

Efficacy and safety of immunological checkpoint inhibitors combined with anti-angiogenic drugs in first-line treatment of metastatic renal cell carcinoma: a systematic review and meta-analysis.

BACKGROUND: Immune checkpoint inhibitors (ICIs) have shown promising results for the second-line treatment of metastatic renal cell carcinoma (mRCC). Several randomized controlled trials have so far also evaluated the efficacy of ICIs for first-line treatment of mRCC. In this study, we conducted a meta-analysis of relevant studies to further clarify the efficacy and safety of ICIs combined with anti-angiogenic drugs for the treatment of mRCC. METHODS: We searched the PubMed, Embase, and Cochrane libraries for RCT trials of ICIs combined with anti-angiogenic drugs for first-line treatment of mRCC published before November 20, 2019. A meta-analysis was conducted based on methodological recommendations by the Cochrane Collaboration. RESULTS: Four articles with a total of 2,967 patients met the inclusion criteria. Our meta-analysis revealed that progression-free survival (PFS) and objective response rate (ORR) were significantly improved in the experimental group while there was no significant difference in overall survival (OS) (HR 0.75, 95% CI: 0.67-0.84; HR 1.43, 95% CI: 1.07-1.91; HR 0.74, 95% CI: 0.53-1.03). After stratification for PD-L1 expression, OS, PFS, and ORR of PD-L1 positive patients were significantly increased in the experimental group (HR 0.74, 95% CI: 0.56-0.96; HR 1.66, 95% CI: 1.11-2.49; HR 0.65, 95% CI: 0.57-0.75). CONCLUSIONS: Immunological checkpoint inhibitors combined with anti-angiogenic drugs as a first-line treatment for mRCC improve PFS and ORR. This effect is more pronounced in PD-L1 positive patients, where ICIs also improve OS. ICIs do not increase the incidence of adverse events.

Establishment of Sunitinib-Resistant Xenograft Model of Renal Cell Carcinoma and the Identification of Drug-Resistant Hub Genes and Pathways.

INTRODUCTION: Sunitinib is the first-line targeted drug for the treatment of advanced renal cell carcinoma (RCC), but its therapeutic potential is limited by premature drug resistance. In an attempt to overcome this limitation, a sunitinib-resistant cell-derived xenograft (CDX) model of clear cell renal cell carcinoma (ccRCC) in vivo was constructed in this study. The molecular mechanism of drug resistance was analyzed using sequencing and bioinformatics tools. METHODS: First, mice were injected subcutaneously with tumor cells 786-O to create tumors and were simultaneously treated with sunitinib. After three consecutive passages, a drug-resistant xenograft model was obtained. Then, key pathways and genes were identified via second-generation sequencing of the tissue and the examination of differentially expressed genes (DEGs) with bioinformatics tools. RESULTS: Analysis of sequencing data revealed that 646 DEGs were upregulated and 465 were downregulated in the drug-resistant tissues when compared with the sensitive tissues. GO showed that the DEGs were significantly enriched in angiogenesis, cell hypoxia response, and apoptosis. KEGG analysis demonstrated that the main pathways were PI3K-Akt, HIF-1, NF-kappa B, and MAPK. Modular analysis of the PPI network indicated that the GO and KEGG analyses of module 1 with the highest ranking were mainly related to ubiquitinase activity. Similarly, the GO and KEGG analyses of the top 10 hub genes were also chiefly linked to ubiquitinase activity. Then, comprehensive expression analysis of the hub genes, and finally, the genes BTRC and TRIM32 were identified, which were consistent in all observations. CONCLUSION: In this study, through the construction of in vitro models and bioinformatics analysis, the important pathways and key genes related to ccRCC sunitinib resistance were discovered. Among them, ubiquitinase may play an important role in drug resistance and may be a potential therapeutic target and biomarker.

Circular RNA circPPP6R3 upregulates CD44 to promote the progression of clear cell renal cell carcinoma via sponging miR-1238-3p.

Circular RNAs (circRNAs) are a type of covalently closed circular-formed RNAs and play crucial roles in the oncogenesis and progression of various human cancers. Here we identified a novel circRNA, circPPP6R3, to be highly expressed both in clear cell renal cell carcinoma (ccRCC) tissues and cell lines based on analyzing high-throughput sequencing data and qRT-PCR analysis. Highly expressed circPPP6R3 was positively correlated with higher histological grade, T stage, and M stage as well as advanced clinical stage of ccRCC patients. Functionally, knockdown of circPPP6R3 attenuated the proliferation, migration, and invasion of ccRCC cells whereas overexpression had the reverse effects. Mechanistically, the biotin-labeled pull-down assay and dual-luciferase reporter assay revealed that circPPP6R3 directly interacted with miR-1238-3p. miR-1238-3p inhibitors had a rescue effect on the proliferative and metastatic capacities by knockdown of circPPP6R3. Moreover, RNA-sequencing analysis and dual-luciferase reporter assay indicated that circPPP6R3 upregulated CD44, a cell-surface glycoprotein contributed to the cell adhesion and metastasis, via sponging to miR-1238-3p. Further investigation revealed that MMP9 and Vimentin were regulated by CD44 in ccRCC. Our study thus provided evidence that the regulatory network involving circPPP6R3/miR-1238-3p/CD44 axis might provide promising biomarkers as well as a therapeutic approach for ccRCC.

[Inhibition and significance of pigment epithelium-derived factor in the development and metastasis of prostate cancer].

OBJECTIVE: To study the inhibition and significance of pigment epithelium-derived factor (PEDF) in the development and metastasis of prostate cancer. METHODS: The expression of PEDF was examined in the normal prostate tissue, benign prostatic hyperplasia, prostate cancer tissue and prostate cancer cell lines, PC-3 and Lncap by immunohistochemical SP method and Western blot. In combination with clinical data, statistical analysis was performed to evaluate the relation of the expression level of PEDF in prostate cancer and the relationship between different histological grades of prostate cancer. RESULTS: In normal prostate tissue and benign prostate tissue, the expression of PEDF were elevated and it was far higher than the prostate cancer and prostate cancer cell line. The expression of PEDF and the pathological grade of prostate cancer were related to the differentiated carcinoma of prostate tissue, and the expression level of PEDF in poorly differentiated carcinoma below the highly differentiated carcinoma of prostate cancer. In metastatic prostate cancer, the expression of PEDF was lower than that of prostate cancer without metastasis (12% vs 43.1%). CONCLUSION: The expression of PEDF and the incidence of prostate cancer have a negative correlation. The lower grade of prostate cancer cells, the less the tissue expression of PEDF and the higher tendency of invasion and metastasis.

[Application of modified single port laparoscopic radical cystoprostatectomy and orthotopic ileal neobladder].

OBJECTIVE: To evaluated the feasibility and therapeutic effect of laparoscopic-endoscopic single-site surgery (LESS) for radical cystoprostatectomy and orthotopic ileal neobladder. METHODS: Between November 2008 and August 2009, 8 male patients with bladder cancer underwent modified single port laparoscopic cystoprostatectomy and orthotopic ileal neobladder with a home-made multichannel port made up of two stretchable rings and a surgical glove with trocars and valves attached to its fingers. A 4 - 5 cm midline incision was made in lower abdomen and the multichannel port placed into this incision. Laparoscopic instrument was extended through the multichannel port. Another port was placed at the umbilicus site for laparoscope. The investigators performed the modified single port transperitoneal technique neobladder extracorporeally and urethro-pouch anastomosis under laparoscopy with a slipknot running suture technique. The perioperative and postoperative data were collected and analyzed prospectively. RESULTS: In all cases, no conversion into conventional laparoscopic or open surgery was necessary. The mean operative duration, including extended lymph node dissection, was 399 min (range: 355 - 455 min). Mean estimated blood loss was 154 ml (range: 90 -210 ml). The mean postoperative hospital stay was 15 d (range: 14 - 18 d). No peri-operative mortality was reported. One case had delirium 2 days postoperatively and there was no port-related complication. The pathological report confirmed surgical margins were tumor-free for all cases. The average number of removed lymph node was 20. The mean follow-up period was 6.1 months (range: 2.0 -10.0 months). All patients remained disease-free until the last follow-up. CONCLUSIONS: With a reduction of 4 ports, modified single port laparoscopic radical cystectomy with orthotopic ileal neobladder is technically feasible with a low complication rate. Our initial outcome is encouraging, but additional work and a further follow-up are warranted to critically compare the oncological outcome with open surgery.

Long-Term Oncologic Outcomes After Laparoscopic and Robotic Tumor Enucleation for Renal Cell Carcinoma.

OBJECTIVES: Tumor enucleation (TE) optimizes parenchymal preservation with promising short-term oncologic outcomes compared with standard partial nephrectomy (SPN). However, researches/literatures about long-term oncologic outcomes for TE after minimally invasive surgery are scarce. We aim to analyze long-term oncologic outcomes after laparoscopic and robotic tumor enucleation for renal cell carcinoma (RCC). PATIENTS AND METHODS: We retrospectively analyzed 146 patients who underwent TE with either laparoscopic or robotic approach for localized RCC in our center. Local recurrence, cancer specific survival (CSS), recurrence free survival (RFS), and overall survival (OS) were the main outcomes. Survival curves were generated using a Kaplan-Meier method. Perioperative outcomes and pathological outcomes were also analyzed. RESULTS: Overall, 98 male and 48 female patients were eligible for the study. The median tumor size was 3.4 cm with a median R.E.N.A.L. score of seven. Warm ischemia was used in 143 patients with a median ischemia time of 20 min and three patients had zero ischemia. Five patients (3.4%) had major complications (> Clavien IIIa) and only two were related to urinary system. The median global glomerular filtration rate (GFR) preserved after surgery was 93%. Pseudocapsule invasion was reported in 50 tumors (34%) and positive surgical margins were found in 3/146 (2.1%) tumors. At a median follow-up of 66 months, local recurrence happened in two patients (1.4%), and systemic recurrence happened in six patients (4.2%). The 5-year CSS, RFS, OS were 95.7, 89.6, and 91.9%, and the 10-year CSS, RFS, OS were 93.8, 89.6, and 90.0%, respectively. CONCLUSION: This study indicates that tumor enucleation with laparoscopic or robotic approach in experienced hands for the treatment of RCC appears oncologically safe with a median follow-up of more than 5 years. Prospective studies with more patients and longer follow-up will be required to further evaluate oncologic safety after TE.

Laparoscopic radical cystectomy with orthotopic ileal neobladder: a report of 85 cases.

PURPOSE: The preliminary results of laparoscopic radical cystectomy in 85 patients are presented in this study. The functional and oncologic outcomes of this procedure in these patients are discussed. PATIENTS AND METHODS: Between December 2002 and May 2006, we performed 85 laparoscopic radical cystectomies with orthotopic ileal neobladder for bladder cancer in 77 men and 8 women. A 5-port transperitoneal approach was applied. The standard bilateral pelvic lymphadenectomy was performed first, then radical cystectomy was completed laparoscopically. The construction of the ileal neobladder and the anastomosis of ureter-neobladder were performed extracorporeally. The neobladder was anastomosed to the urethral stump under laparoscopy. A nerve-sparing procedure was performed for eight patients. RESULTS: The median operative time was 320 min, and the median blood loss was 280 mL. Conversion to open surgery was not necessary in any of the patients. The average time to oral intake after operation was 3.9 days. There were no perioperative mortalities. The complication rate was 14.1% (12/85), including such complications as three uretero-pouch anastomotic strictures, one vesicourethral anastomotic stricture, one pouch-vaginal fistula, one colonic pouch fistula, one ileo-pouch fistula, three ileus, one pneumonia, and one pyelonephritis. The daytime continence rate was 91.2%, and the nighttime continence rate was 82.4% at 6 months postoperatively. The neobladder capacity was about 343 mL. Surgical margins were tumor free for all patients. Of the eight patients who underwent a nerve-sparing procedure, four patients had potency for intercourse. During a follow-up period of 1 to 41 months (average 21.3 months), three patients had local recurrence, one patient had trocar site seeding, and five patients had distant metastasis, of whom four died. CONCLUSIONS: Laparoscopic radical cystectomy with extracorporeal formation of a neobladder is a feasible procedure with low morbidity and acceptable neobladder function. Long-term follow-up is needed to confirm the oncologic outcomes.