Characterization of Uptake and Internalization of Exosomes by Bladder Cancer Cells

Bladder tumors represent a special therapeutic challenge as they have a high recurrence rate requiring repeated interventions and may progress to invasive or metastatic disease. Exosomes carry proteins implicated in bladder cancer progression and have been implicated in bladder cancer cell survival. Here, we characterized exosome uptake and internalization by human bladder cancer cells using Amnis ImageStreamX, an image cytometer. Exosomes were isolated by ultracentrifugation from bladder cancer culture conditioned supernatant, labeled with PKH-26, and analyzed on the ImageStreamX with an internal standard added to determine concentration. Exosomes were cocultured with bladder cancer cells and analyzed for internalization. Using the IDEAS software, we determined exosome uptake based on the number of PKH-26+ spots and overall PKH-26 fluorescence intensity. Using unlabeled beads of a known concentration and size, we were able to determine concentrations of exosomes isolated from bladder cancer cells. We measured exosome uptake by recipient bladder cancer cells, and we demonstrated that uptake is dose and time dependent. Finally, we found that uptake is active and specific, which can be partially blocked by heparin treatment. The characterization of cellular uptake and internalization by bladder cancer cells may shed light on the role of exosomes on bladder cancer recurrence and progression.

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Leupaxin Promotes Bladder Cancer Proliferation, Metastasis, and Angiogenesis Through the PI3K/AKT Pathway

Cellular Physiology and Biochemistry ◽

10.1159/000491536 ◽

2018 ◽

Vol 47 (6) ◽

pp. 2250-2260 ◽

Cited By ~ 5

Author(s):

Teng Hou ◽

Lijie Zhou ◽

Longwang Wang ◽

Gallina Kazobinka ◽

Yumao Chen ◽

...

Keyword(s):

Bladder Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Akt Pathway ◽

Advanced Tumor Stage ◽

Large Tumor ◽

Advanced Tumor ◽

Bladder Cancer Cells ◽

The Impact

Background/Aims: Leupaxin (LPXN) is a member of the paxillin protein family. Several studies have reported that LPXN regulates cancer development; however, the role of LPXN in bladder cancer remains unknown. Methods: The expression of LPXN in bladder cancer cells and tissues was determined by real-time PCR, western blotting, and immunohistochemistry, respectively. The biological role of LPXN in bladder cancer cell proliferation, invasion, and angiogenesis was explored both in vitro and in vivo. Results: LPXN expression was elevated in bladder cancer tissues and cell lines compared to adjacent non-tumor tissues and normal urothelial cells. High LPXN expression was correlated with large tumor size, advanced tumor stage, and poor survival in bladder cancer patients. Overexpression of LPXN significantly promoted the proliferation, invasion, and angiogenesis of bladder cancer cells, while suppressing LPXN had the opposite effects. The impact on tumor progression was abolished by inhibiting PI3K/ AKT signaling pathway. We further demonstrated that LPXN probably up-regulated S100P via the PI3K/AKT pathway. Conclusions: LPXN may facilitate bladder cancer progression by upregulating the expression of S100P via PI3K/AKT pathway. These results provide a novel insight into the role of LPXN in tumorigenesis and progression of bladder cancer and potential therapeutic target of bladder cancer.

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LINC00958 promotes bladder cancer carcinogenesis by targeting miR-490-3p and AURKA

BMC Cancer ◽

10.1186/s12885-021-08882-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Hongtao Zhen ◽

Peng Du ◽

Qiang Yi ◽

Xiaolong Tang ◽

Tongqing Wang

Keyword(s):

Bladder Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Treatment Method ◽

Bladder Cancer Cells ◽

Cell Progression ◽

New Treatment ◽

Cancer Tissues ◽

Proliferation And Invasion

Abstract Background Bladder cancer is a prevalent malignancy of the urinary system, in which long non-coding RNAs (lncRNAs) are highly associated. We aimed to elucidate the role of LINC00958 in bladder cancer. Methods LINC00958 expression levels were measured using qRT-PCR. The interaction of LINC00958-miR-490-3p-AURKA was analyzed by luciferase, RIP, and RNA pull-down assays. The biological roles of LINC00958, miR-490-3p, and AURKA in bladder cancer cells were analyzed using CCK8, BrdU, and transwell assays. Results Increased expression of LINC00958 and AURKA was observed in bladder cancer tissues and cell lines. Decreased LINC00958 expression repressed bladder cancer progression and downregulation of miR-490-3p accelerated bladder cancer cell progression. Moreover, LINC00958 sponges miR-490-3p to upregulate AURKA expression, thereby promoting carcinogenesis in bladder cancer cells. Conclusions Our study revealed that LINC00958 facilitated cell proliferation and invasion, and suppressed cell apoptosis by sponging miR-490-3p and upregulating AURKA, thus inspiring a new treatment method for bladder cancer.

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USP24-GSDMB complex promotes bladder cancer proliferation via activation of the STAT3 pathway

10.21203/rs.3.rs-95101/v1 ◽

2020 ◽

Author(s):

Haiqing He ◽

Bin Zhang ◽

Bin Yan ◽

Ming Xiao ◽

Jiannan Ren ◽

...

Keyword(s):

Bladder Cancer ◽

Cancer Cells ◽

Cancer Progression ◽

Annexin V ◽

Cell Counting ◽

Cancer Proliferation ◽

Data Set ◽

Stat3 Signaling ◽

Bladder Cancer Cells

Abstract BackgroundBladder cancer is the fourth and tenth most common malignancy in men and women worldwide, respectively. One of the main reasons for the unsatisfactory therapeutic control of bladder cancer is that the molecular biological mechanism of bladder cancer is complex. Gasdermin B (GSDMB) is one member of the gasdermin family and participates in the regulation of cell pyroptosis. The role of GSDMB in bladder cancer has not been studied to date.MethodsThe clinical relevance of GSDMB was examined by the TCGA data set. Functional assays, such as the MTT assay, Celigo fluorescent cell-counting assay, Annexin V-APC assay and xenografts, were used to determine the biological role of GSDMB in bladder cancer. The interaction between GSDMB and STAT3, or GSDMB and USP24 were detected by Mass spectrometry and verified through immunoprecipitation. The relationship between USP24, GSDMB and STAT3 was examined by Western blot analysis and immunohistochemistry.ResultsIn this study, bioinformatics analysis indicated that the mRNA expression level of GSDMB in bladder cancer tissues was higher than that in adjacent normal tissues. Then, we showed that GSDMB promoted bladder cancer progression. Furthermore, we demonstrated that GSDMB interacted with STAT3 to increase the phosphorylation of STAT3 and modulate the glucose metabolism and promote tumor growth in bladder cancer cells. Besides, we also showed that USP24 stabilized GSDMB to activate STAT3 signaling, which was blocked by the USP24 inhibitor. ConclusionsWe suggested that aberrantly up-regulated GSDMB was responsible for enhancing the growth and invasion ability of bladder cancer cells. Then, we showed that GSDMB could bind to STAT3 and activate STAT3 signaling in bladder cancer. Furthermore, we also demonstrated that USP24 interacts with GSDMB and prevents GSDMB from degradation in bladder cancer cells. Therefore, the USP24/GSDMB/STAT3 axis may be a new targetable signaling pathway for bladder cancer treatment.

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792: Role of Oxidative Stress in Tumorigenic Potential of Bladder Cancer Cells

The Journal of Urology ◽

10.1016/s0022-5347(18)34961-9 ◽

2005 ◽

Vol 173 (4S) ◽

pp. 214-215 ◽

Cited By ~ 2

Author(s):

Daniel Cho ◽

Xiao Fang Ha ◽

J. Andre Melendez ◽

Louis J. Giorgi ◽

Badar M. Mian

Keyword(s):

Oxidative Stress ◽

Bladder Cancer ◽

Cancer Cells ◽

Tumorigenic Potential ◽

Bladder Cancer Cells

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514 Role of macroautophagy in the response of bladder cancer cells to the pan Bcl-2 inhibitor AT-101 (Gossypol)

European Urology Supplements ◽

10.1016/s1569-9056(14)60506-6 ◽

2014 ◽

Vol 13 (1) ◽

pp. e514

Author(s):

J. Mani ◽

S. Vallo ◽

S. Rakel ◽

P. Antonietti ◽

G. Bartsch ◽

...

Keyword(s):

Bladder Cancer ◽

Cancer Cells ◽

Bladder Cancer Cells

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ROLE OF MITOCHONDRIA IN CIPROFLOXACIN INDUCED APOPTOSIS IN BLADDER CANCER CELLS

The Journal of Urology ◽

10.1016/s0022-5347(05)65283-4 ◽

2002 ◽

Vol 167 (3) ◽

pp. 1288-1294 ◽

Cited By ~ 21

Author(s):

OLIVIA ARANHA ◽

LIPING ZHU ◽

SAMIR ALHASAN ◽

DAVID P. WOOD ◽

TUAN H. KUO ◽

...

Keyword(s):

Bladder Cancer ◽

Cancer Cells ◽

Bladder Cancer Cells ◽

Induced Apoptosis

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The role of cyclooxygenase enzymes in the growth of human gall bladder cancer cells

Carcinogenesis ◽

10.1093/carcin/21.5.403 ◽

2000 ◽

Vol 21 (7) ◽

pp. 1403-1409 ◽

Cited By ~ 4

Author(s):

Erik M. Grossman ◽

Walter E. Longo ◽

Ninder Panesar ◽

John E. Mazuski ◽

Donald L. Kaminski

Keyword(s):

Bladder Cancer ◽

Gall Bladder ◽

Cancer Cells ◽

Gall Bladder Cancer ◽

Bladder Cancer Cells

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Pathogenic and Diagnostic Potential of BLCA-1 and BLCA-4 Nuclear Proteins in Urothelial Cell Carcinoma of Human Bladder

Advances in Urology ◽

10.1155/2012/397412 ◽

2012 ◽

Vol 2012 ◽

pp. 1-5 ◽

Cited By ~ 8

Author(s):

Matteo Santoni ◽

Francesco Catanzariti ◽

Daniele Minardi ◽

Luciano Burattini ◽

Massimo Nabissi ◽

...

Keyword(s):

Bladder Cancer ◽

Cell Carcinoma ◽

Matrix Protein ◽

Transitional Cell ◽

High Recurrence Rate ◽

Diagnostic Potential ◽

Bladder Cancer Cells ◽

Previously Treated Patients ◽

Previously Treated

Transitional cell carcinoma (TCC) of the bladder is one of the most common malignancies of genitourinary tract. Patients with bladder cancer need a life-long surveillance, directly due to the relatively high recurrence rate of this tumor. The use of cystoscopy represents the gold standard for the followup of previously treated patients. Nevertheless, several factors, including cost and invasiveness, render cystoscopy not ideal for routine controls. Advances in the identification of specific alterations in the nuclear structure of bladder cancer cells have opened novel diagnostic landscapes. The members of nuclear matrix protein family BLCA-1 and BLCA-4, are currently under evaluation as bladder cancer urinary markers. They are involved in tumour cell proliferation, survival, and angiogenesis. In this paper, we illustrate the role of BLCA-1 and BLCA-4 in bladder carcinogenesis and their potential exploitation as biomarkers in this cancer.

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Exosome-Derived LINC00960 and LINC02470 Promote the Epithelial-Mesenchymal Transition and Aggressiveness of Bladder Cancer Cells

Cells ◽

10.3390/cells9061419 ◽

2020 ◽

Vol 9 (6) ◽

pp. 1419

Author(s):

Cheng-Shuo Huang ◽

Jar-Yi Ho ◽

Jung-Hwa Chiang ◽

Cheng-Ping Yu ◽

Dah-Shyong Yu

Keyword(s):

Bladder Cancer ◽

Notch Signaling ◽

Cancer Cells ◽

Signaling Pathways ◽

Cancer Progression ◽

Epithelial Mesenchymal Transition ◽

Low Grade ◽

High Grade ◽

Mesenchymal Transition ◽

Bladder Cancer Cells

Exosomes are essential for several tumor progression-related processes, including the epithelial–mesenchymal transition (EMT). Long non-coding RNAs (lncRNAs) comprise a major group of exosomal components and regulate the neoplastic development of several cancer types; however, the progressive role of exosomal lncRNAs in bladder cancer have rarely been addressed. In this study, we identified two potential aggressiveness-promoting exosomal lncRNAs, LINC00960 and LINC02470. Exosomes derived from high-grade bladder cancer cells enhanced the viability, migration, invasion and clonogenicity of recipient low-grade bladder cancer cells and activated major EMT-upstream signaling pathways, including β-catenin signaling, Notch signaling, and Smad2/3 signaling pathways. Nevertheless, LINC00960 and LINC02470 were expressed at significantly higher levels in T24 and J82 cells and their secreted exosomes than in TSGH-8301 cells. Moreover, exosomes derived from LINC00960 knockdown or LINC02470 knockdown T24 cells significantly attenuated the ability of exosomes to promote cell aggressiveness and activate EMT-related signaling pathways in recipient TSGH-8301 cells. Our findings indicate that exosome-derived LINC00960 and LINC02470 from high-grade bladder cancer cells promote the malignant behaviors of recipient low-grade bladder cancer cells and induce EMT by upregulating β-catenin signaling, Notch signaling, and Smad2/3 signaling. Both lncRNAs may serve as potential liquid biomarkers for the prognostic surveillance of bladder cancer progression.

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