Selinexor demonstrates anti-tumor efficacy in paired patient-derived xenograft models and hydrogel-embedded histoculture drug sensitivity test of penile cancer.

BACKGROUND: Penile cancer is a rare malignancy with a poor prognosis, even with various treatment options. Considering the little progress in the study of the pathogenesis and treatment of penile cancer because of the lack of models that mimic the biological properties of the tumor, we have developed a patient-derived xenograft (PDX) model and paired hydrogel-embedded histoculture drug sensitivity test (HDST) to screen for drugs that can inhibit tumors. The increased expression of XPO1, as a key nuclear export protein involved in the transport of various tumor suppressors and cell cycle regulatory proteins, is associated with the prognosis of a variety of tumors [World J Uroly 27(2):141-150, 2009]. Selinexor is an inhibitor of XPO1, which can treat cancers, such as multiple myeloma, gastric cancer, triple-negative breast cancer, and non-small cell carcinoma [Transl Androl Urol 6(5):785-790, 2017; OncoTargets Therapy 13:6405-6416, 2020]. However, whether XPO1 inhibition has a role in penile cancer remains unknown. Therefore, this article used the PDX and HDST models to investigate whether the inhibition of XPO1 has an effect on penile cancer and its underlying mechanism. METHODS: We used penile cancer tumor tissues to construct a PDX model of penile cancer and paired PDXE model and confirmed the consistency of PDX tumor tissues in source patients. Then, we assessed the ability of Selinexor to inhibit penile cancer tissues in vivo using a PDX model and in vitro by HDST. We also examined the potential mechanism of XPO1 action on penile cancer by IHC and TUNEL. Finally, we assessed the safety of the drug treatment by H&E and biochemical blood analysis. RESULTS: Result showed that the penile cancer PDX model and patient penile cancer tissues were clinically consistent in morphological characteristics and protein expression. In addition, Selinexor could inhibit tumor growth in PDX models and HDST. We found that P53, P21 expression was upregulated; Cyclin D1 expression was downregulated, and apoptosis of tumor cells was increased in the Selinexor-treated PDX model. Moreover, it had no significant effect on liver, kidney, and cardiac function. CONCLUSION: The PDX model of penile cancer was a powerful tool for penile cancer research and new drug development. It showed that Selinexor can effectively inhibit penile cancer in vitro and in vivo. In addition, XPO1 may affect P53, P21, and Cyclin D1 expression to regulate the growth and apoptosis of penile carcinoma.

Genome-wide screening for the G-protein-coupled receptor (GPCR) pathway-related therapeutic gene RGS19 (regulator of G protein signaling 19) in bladder cancer.

Bladder cancer is one of the most severe genitourinary cancers, causing high morbidity worldwide. However, the underlying molecular mechanism is not clear, and it is urgent to find target genes for treatment. G-protein-coupled receptors are currently a target of high interest for drug design. Thus, we aimed to identify a target gene-related to G-protein-coupled receptors for therapy. We used The Cancer Genome Atlas (TCGA) and DepMap databases to obtain the expression and clinical data of RGS19. The results showed that RGS19 was overexpressed in a wide range of tumor, especially bladder cancer. We also explored its effect on various types of cancer. High expression of RGS19 was also shown to be significantly associated with poor prognosis. Cell models were constructed for cell cycle detection. shRGS19 can halt the cell cycle at a polyploid point. RGS19 is a G-protein-coupled receptor signaling pathway-related gene with a significant effect on survival. We chose RGS19 as a therapeutic target gene in bladder cancer. The drug GSK1070916 was found to inhibit the effect of RGS19 via cell rescue experiments in vitro.

Endovascular removal of a misplaced ureteral stent in the vena cava: a complication of ultrasound-guided percutaneous nephrolithotomy.

Intravascular migration of a double J stent into the inferior vena cava is an uncommon complication. Active prevention, timely diagnosis, and early intervention are crucial for this complication. Intravascular interventional therapy is relatively easy, less traumatic, and has a high success rate. It can be used to select patients for intravascular ectopic DJS treatment.

FAT10 promotes the progression of bladder cancer by upregulating HK2 through the EGFR/AKT pathway.

The ubiquitin-like protein FAT10 and the hexokinase protein HK2 play vital regulatory roles in several cellular processes. However, the relationship between these two proteins and their role in the pathogenesis of bladder cancer are not well understood. Here, we found that FAT10 and HK2 protein levels were markedly higher in bladder cancer tissues than in normal adjacent tissues. In addition, RNAi-mediated silencing of FAT10 led to reduced HK2 levels and suppressed bladder cancer progression in vivo and in vitro. The results of our in vivo and in vitro experiments revealed that HK2 is critical for FAT10-mediated progression of bladder cancer. The current study demonstrated that FAT10 enhanced the progression of bladder cancer by positively regulating HK2 via the EGFR/AKT pathway. Based on our findings, FAT10 is believed to stabilize EGFR expression by modulating its degradation and ubiquitination. The results of the current study indicate that there is a correlation between FAT10 and HK2 in the progression of bladder cancer. In addition, we identified a new pathway that may be involved in the regulation of HK2. These findings implicate dysfunction of the FAT10, EGFR/AKT, and HK2 regulatory circuit in the progression of bladder cancer.

PRMT7 promotes the growth of renal cell carcinoma through modulating the ß-catenin/C-MYC axis.

Arginine methylation is mainly catalyzed by protein arginine methyltransferases (PRMTs) and is one of the most common posttranslational modifications closely related to the development of cancer. PRMT7 is overexpressed in various tumors and promotes the malignant progression of tumors, but the expression and role of PRMT7 in renal cell carcinoma (RCC) remains unclear. Here, we report for the first time that the expression of PRMT7 is increased in clear cell renal cell carcinoma (ccRCC) tissues and that it may act as an independent predictor for the poor prognosis of ccRCC patients. We found that PRMT7 promotes RCC cell proliferation both in vitro and in vivo. Moreover, the methyltransferase inhibitor adenosine dialdehyde (Adox) blocks the action of PRMT7 in ccRCC cells. Furthermore, PRMT7 regulates the expression of C-MYC, which plays an important role in promoting ccRCC cell proliferation, and it accelerates the tumor development of RCC in a C-MYC-dependent manner. Mechanistically, PRMT7 upregulates the expression of C-MYC via methylating ß-catenin and inhibiting the ubiquitin-mediated degradation of ß-catenin. In conclusion, our study demonstrates that overexpressed PRMT7 in ccRCC cells acts as an oncogene to promote the growth of renal cell carcinoma through regulating the ß-catenin/C-MYC axis, thereby providing new strategies and targets for the treatment of ccRCC patients.

Circular RNA circ-VANGL1 as a competing endogenous RNA contributes to bladder cancer progression by regulating miR-605-3p/VANGL1 pathway.

Increasing reports indicate that circular RNAs (circRNAs) are very important regulators in human diseases, including cancers. In bladder cancer (BC), several circRNAs have been reported to be involved in tumor progressions, such as circ-ITCH and circTCF25. However, the functions of most circRNAs in BC still remains largely unknown. In this study, we identified a novel circRNA termed as circ-VANGL1 by bioinformatics analysis. We found that circ-VANGL1 was highly expressed in BC tissues compared with adjacent normal tissues. Furthermore, we showed that circ-VANGL1 could serve as a prognostic marker for patients with BC. Through functional experiments, we found that circ-VANGL1 knockdown significantly suppressed BC cell proliferation, cell cycle, migration, and invasion in vitro. Besides, circ-VANGL1 silence inhibited BC cell propagation in vivo. Mechanistically, we identified circ-VANGL1 as a sponge of miR-605-3p which targeted VANGL1 in BC cells. Through repressing miR-605-3p availability, circ-VANGL1 contributes to VANGL1 expression, consequently leading to BC cell proliferation, migration, and invasion. Taken together, our study demonstrated circ-VANGL1/miR-605-3p/VANGL1 as a novel essential signaling pathway involved in BC progression.

Urothelium with barrier function differentiated from human urine-derived stem cells for potential use in urinary tract reconstruction.

BACKGROUND: Autologous urothelial cells are often obtained via bladder biopsy to generate the bio-engineered urethra or bladder, while urine-derived stem cells (USC) can be obtained by a non-invasive approach. The objective of this study is to develop an optimal strategy for urothelium with permeability barrier properties using human USC which could be used for tissue repair in the urinary tract system. METHODS: USC were harvested from six healthy adult individuals. To optimize urothelial differentiation, five different differentiation methods were studied. The induced cells were assessed for gene and protein expression markers of urothelial cells via RT-PCR, Western blotting, and immunofluorescent staining. Barrier function and ultrastructure of the tight junction were assessed with permeability assays and transmission electron microscopy (TEM). Induced cells were both cultured on trans-well membranes and small intestinal submucosa, then investigated under histology analysis. RESULTS: Differentiated USC expressed significantly higher levels of urothelial-specific transcripts and proteins (Uroplakin III and Ia), epithelial cell markers (CK20 and AE1/AE3), and tight junction markers (ZO-1, ZO-2, E-cadherin, and Cingulin) in a time-dependent manner, compared to non-induced USC. In vitro assays using fluorescent dye demonstrated a significant reduction in permeability of differentiated USC. In addition, transmission electron microscopy confirmed appropriate ultrastructure of urothelium differentiated from USC, including tight junction formation between neighboring cells, which was similar to positive controls. Furthermore, multilayered urothelial tissues formed 2 weeks after USC were differentiated on intestine submucosal matrix. CONCLUSION: The present study illustrates an optimal strategy for the generation of differentiated urothelium from stem cells isolated from the urine. The induced urothelium is phenotypically and functionally like native urothelium and has proposed uses in in vivo urological tissue repair or in vitro urethra or bladder modeling.

Human Urine-Derived Stem Cell Differentiation to Endothelial Cells with Barrier Function and Nitric Oxide Production.

Endothelial cells (ECs) play a key role in revascularization within regenerating tissue. Stem cells are often used as an alternative cell source when ECs are not available. Several cell types have been used to give rise to ECs, such as umbilical cord vessels, or differentiated from somatic stem cells, embryonic, or induced pluripotent stem cells. However, the latter carry the potential risk of chronic immune rejection and oncogenesis. Autologous endothelial precursors are an ideal resource, but currently require an invasive procedure to obtain them from the patient's own blood vessels or bone marrow. Thus, the goal of this study was to determine whether urine-derived stem cells (USCs) could differentiate into functional ECs in vitro. Urine-derived cells were then differentiated into cells of the endothelial lineage using endothelial differentiation medium for 14 days. Changes in morphology and ultrastructure, and functional endothelial marker expression were assessed in the induced USCs in vitro. Grafts of the differentiated USCs were then subcutaneously injected into nude mice. Induced USCs expressed significantly higher levels of specific markers of ECs (CD31, vWF, eNOS) in vitro and in vivo, compared to nondifferentiated USCs. In addition, the differentiated USC formed intricate tubular networks and presented similar tight junctions, and migration and invasion ability, as well as ability to produce nitric oxide (NO) compared to controls. Using USCs as autologous EC sources for vessel, tissue engineering strategies can yield a sufficient number of cells via a noninvasive, simple, and low-cost method suitable for rapid clinical translation. Stem Cells Translational Medicine 2018 Stem Cells Translational Medicine 2018;7:686-698.

Fibroblasts from pBOO promote tumorigenesis by secreting TGFbeta1 to induce EMT in bladder urothelial carcinoma cells.

Urinary bladder cancer (UBC) is one of the most common malignancies worldwide. UBC patients at muscle invasive stage have poor clinical outcome, due to high propensity for metastasis. Non-tumor activated fibroblasts, named α-SMA+Fs, is similar to carcinoma-associated fibroblasts (CAFs) which could express α-SMA. However, whether α-SMA+Fs patients could induce UBC cell invasion is unclear. Herein, we found that characterization of primary α-SMA+Fs separated from PBOO (partial bladder outlet obstruction) rats was fell in between normal fibroblasts (α-SMA-Fs) and CAFs. Additionally, the conditional medium from α-SMA+Fs enhanced the NBT-II cell invasion through inducing EMT, and the oncogenic function of mixed supernatant of α-SMA+Fs/CAFs was stronger than that of CAFs. Inhibition of TGF-ß1 by TGF-ß1 neutralizing antibody decreased the EMT-associated gene expression and NBT-II cell invasion, suggesting that α-SMA+Fs can induce tumor EMT through TGF-ß1. Xenograft experiments showed that the tumorigenic effect of α-SMA+Fs in mice was also between CAFs and α-SMA-Fs, and α-SMA+Fs/CAFs also had a strong tumorigenic effect. We preformed rats with PBOO and found that the incidence of invasive bladder cancer in PBOO+BBN group was higher than in BBN group, suggesting the PBOO treatment contributed to tumorigenesis. Thus, α-SMA+Fs promoted tumorigenesis by secreting TGF-ß1 to induce EMT.

Ectopic insertion of a duplicated ureter into prostatic urethra: Demonstration by 3D multi-detector computed tomography urography.

Ectopic insertion of the ureter in the genitourinary tract is a rare congenital disorder, usually associated with ureteral duplication. Identification of the insertion open is critical for ureteric re-implantation. However, the challenge in the diagnosis of ectopic insertion of the ureter usually is to identify its insertion, particularly when the affected ureter is not dilated. Multi-detector computed tomography (MDCT) urography with nonionic iodinated contrast media delineates the ureteric course in the normal functioning kidney in the excretory phase [1]. This report presented a young male patient with ectopic insertion of a duplicated ureter diagnosed by MDCT urography. Three-dimensional (3D) analysis technology, such as volume rendering (with a color display improving the visualization of complex anatomy and 3D relationships) and maximum intensity projection (similar in principle to projection angiography), is useful for the illustration of urinary tract anatomy [1]. Rotated volume rendering reconstruction images and continual thinner maximum intensity projection reformatted images can be viewed as videos, which provides detail delineation of the ectopic ureteral insertion and its associated ureteral duplication.In this study, we reported MDCT urography and 3D analysis technology as an appropriate diagnostic method for the ectopic ureteral insertion and its associated complications.

ARHGAP24 inhibits cell cycle progression, induces apoptosis and suppresses invasion in renal cell carcinoma.

Renal cell carcinoma (RCC) is the major cause of kidney malignancy-related deaths. Rho GTPases are key regulators in cancer cell metastasis. ARHGAP24, a Rac-specific member of the Rho GTPase-activating protein family, acts as a functional target of cancer cell migration and invasion. In the present study, we identified ARHGAP24 expression is downregulated in renal cancer tissues and is highly correlated with long-term survival in RCC patients. Therefore, we investigated the biological functions of ARHGAP24 in renal cancer cells. Ectopic expression of ARHGAP24 resulted in inhibited cell proliferation and arrested cell cycle in two renal cancer cell lines (786-0 and Caki-2); the results were confirmed by ARHGAP24 knocking down. In addition, ARHGAP24 significantly reduced the cell invasion ability and induced apoptosis in renal cancer cells. In addition, overexpressing ARHGAP24 impaired tumor formation in vivo. In summary, our results illustrated that ARHGAP24 plays a unique role in RCC progression as a tumor repressor.

Noncontrast multidetector-row computed tomography scanning for detection of radiolucent calculi in acute renal insufficiency caused by bilateral ureteral obstruction of ceftriaxone crystals.

Noncontrast computed tomography (CT) has great advantage with higher sensitivity and more clear modalities in detecting urinary tract radiolucent calculi in patients with acute renal insufficiency (ARI) compared to other image diagnosis approaches. We report two cases (female, 28 years old; male, 39 years old) with persistent flank pain and acute anuria after the administration of ceftriaxone (4.0 g daily) for 2 days intravenously. No abnormality was found in the kidney-ureter- bladder (KUB) areas with plain abdomen X-rays. A diagnosis of bilateral hydronephrosis was made by ultrasound examination in both cases. Serum creatinine levels reached up to 257 and 810 µ mol/L (normal serum creatinine level is 40-130 µ mol/L), respectively. Vague density spots were noticed in the pelvis with noncontrast multidetector-row CT (MDCT) scanning. However, distinguishable clusters of high-density shadows were seen in pelvic areas with maximum intensity projections (MIP, CT values in 30-128 HU). Ceftriaxone crystal calculi were found on both sides of distal ureters under endoscopy. Renal function recovered in both patients after double-J ureteral stents were installed. Out results demonstrated that noncontrast MDCT scanning and MIP reconstruction as an effective diagnostic tool could provide clear images in detection of radiolucent calculi in urinary tract when conventional X-rays image are not suitable in the patients with obstructive anuria and ARI of unknown origin.

[Expression of lymphotactin in renal tuberculosis].

AIM: To study the expression of lymphotactin in normal kidney and renal tuberculosis and the arrangement of lymphotactin and infiltrating CD4(+) T cells and CD8(+) T cells in renal tuberculosis. METHODS: HLptn cDNA of tissues from six cases with normal kidneys and ten cases with tuberculous kidneys was amplified by RT-PCR. The RT-PCR products were separated with 2% of gel. The cDNA was cloned to vector pGM-T Easy and was completely sequenced. The immunohistochemical staining method was used to examine the expression of lymphotactin in normal kidneys and tuberculous kidneys and the expression of CD4 and CD8 in tuberculous kidneys. RESULTS: The sequence of cloned hLptn cDNA was confirmed and it was identical with the sequence of NO.U23772 published in GenBank. Both normal kidneys and tuberculous kidneys expressed hLptn mRNA. HLptn was detected not only in the cells of normal renal glomerulus and renal tubule but also in the cells of remaining renal glomerulus and renal tubule of tuberculous kidneys. The cells expressing surface antigens CD4 and CD8 scattered in granulomas. CONCLUSION: The constructive expression of hLptn is in the cells of renal glomerulus and renal tubule of normal kidney and tuberculous kidney. The accumulation of CD4(+) T cells and CD8(+) T cells in granulomas may not depend on hLptn.

[Preliminary study on lymphotactin expression of monocyte-derived dendritic cells].

AIM: To induce dendritic cells (DCs) from human peripheral blood monocytes in vitro and to investigate the kinetics lymphotactin (Lptn) mRNA expression. METHODS: Monocytes were isolated from normal human peripheral blood cells by density gradient centrifugation and the plastic-adherent cells were cultured with the cytokines (rhGM-CSF, rhIL-4, rhTNF-alpha). The immature and mature DCs' surface antigens CD1a and CD83 were analyzed by flow cytometry (FCM). The morphology of mature DCs induced by rhTNF-alpha was observed under electron microscope. Lptn cDNA was amplified by RT-PCR, cloned to vector pGM-T Easy and completely sequenced. The RT-PCR products of cells were separated on gel and analyzed by semi-quantification. RESULTS: The cells cultured for 7 d exhibited special dendritic morphology of mature DCs and highly expressed CD83. The sequence of cDNA was confirmed and was identical to that of NO. U23772 published in GenBank. The Lptn mRNA expression of DCs was stronger in DCs cultured for 7 d than in those for 5 d, whereas it was not detected in those for 3 d. CONCLUSION: The monocyte-derived dendritic cells can express Lptn mRNA in a maturation-dependent manner.

[Preparation and feasibility of superparamagnetic dextran iron oxide nanoparticles as gene carrier].

BACKGROUND & OBJECTIVE: Application of magnetic nano- particles as gene carrier in gene therapy of tumor has developed quickly. To obtain a new type non-viral gene introduction and therapy system,which is convenient,and can drive target gene to express highly and stably,this study was designed to explore the preparation of superparamagnetic dextran iron oxide nanoparticles(SDION),and the feasibility of SDION used as gene carrier in vitro. METHODS: SDION were prepared by chemical co-precipitation,separated by gel filtration chromatography on Sephacryl S-300HR,and centrifugation techniques,characterized by transmission electron microscopy,laser scattering system,and vibrating sample magnetometer signal processor. The green fluorescent protein-C2 (GFP-C2) plasmid was used as target gene. SDION-GFP- C2 compounds were synthesized by oxidation-reduction reaction. The connection rate of SDION and GFP-C2 was analyzed by agarose electrophoresis,and evaluated by measuring concentration of GFP in the supernatant after centrifugation. Liposome transfection was used as control,the efficiencies of SDION and liposome in transferring GFP gene into human bladder cancer BIU-87 cells were evaluated under fluorescence microscope in vitro. RESULTS: The diameter of SDION ranged from 3 nm to 8 nm, the effective diameter was 59.2 nm, and the saturation magnetization was 0.23 emu/g. After oxidized by sodium periodate of 10 mmol/L,and deoxidized by sodium hydride boron of 0.5 mol/L, SDION could connect with GFP in maximum degree,the transfection efficiency of SDION as gene carrier was about 45%, even higher than that of liposome (about 30%). CONCLUSION: SDION could connect with GFP plasmid by oxidation- reduction reaction,and success to transfer GFP gene into human bladder cancer BIU-87 cells in vitro.