Proliferation Of Bladder Carcinoma Cells Research Articles

Circular RNA circFLNA inhibits the development of bladder carcinoma through microRNA miR-216a-3p/BTG2 axis

ABSTRACT Recent studies have shown that circular RNA circFLNA is abnormally expressed in a variety of malignant tumors, but its role and mechanism in bladder carcinoma (BCa) are still unclear. The present paper aims to contribute to research on the effects and mechanism of circFLNA on the malignant phenotype of BCa. In this study, the expressions of circFLNA, miR-216a-3p and BTG2 in BCa and BCa cells (EJ, T24, 5637, TCC-SUP) were detected by qRT-PCR. EdU staining, colony formation, Transwell assay, wound healing assays, and sphere formation assay were used to measure the cell proliferation, viability, invasion, migration, and cell stemness of BCa cells after circFLNA overexpression. In addition, the correlation existed between miR-216a-3p and circFLNA or BTG2 was confirmed by Dual-Luciferase Reporter assay and RNA pull-down. Western blot was utilized to determine the expression of BTG2, MMP2, epithelial-mesenchymal transition (EMT)-related proteins (vimentin, E-cadherin) and stem cell-specific proteins (CD34, OCT4, SOX2). Our study confirmed that downregulated circFLNA and BTG2 expression and upregulated miR-216a-3p were found in both BCa tissues and cell lines. Meanwhile, upregulated circFLNA inhibited proliferation, invasion and migration, EMT and stemness of BCa cells. MiR-216a-3p was a target gene of circFLNA and could target BTG2. Further analysis finally demonstrated that circFLNA sponged miR-216a-3p and indirectly promoted BTG2 expression, ultimately regulating proliferation, migration, invasion and EMT of BCa cells. In conclusion, circFLNA inhibits the malignant phenotype of BCa cells and their stemness through miR-216a-3p/BTG2, thus suppressing BCa progression.

UBIAD1 suppresses the proliferation of bladder carcinoma cells by regulating H-Ras intracellular trafficking via interaction with the C-terminal domain of H-Ras

UbiA prenyltransferase domain-containing protein 1 (UBIAD1) plays a key role in biosynthesis of vitamin K2 and coenzyme Q10 using geranylgeranyl diphosphate (GGPP). However, the mechanism by which UBIAD1 participates in tumorigenesis remains unknown. This study show that UBIAD1 interacts with H-Ras, retains H-Ras in the Golgi apparatus, prevents H-Ras trafficking from the Golgi apparatus to the plasma membrane, blocks the aberrant activation of Ras/MAPK signaling, and inhibits the proliferation of bladder cancer cells. In addition, GGPP was required to maintain the function of UBIAD1 in regulating the Ras/ERK signaling pathway. A Drosophila model was employed to confirm the function of UBIAD1/HEIX in vivo. The activation of Ras/ERK signaling at the plasma membrane induced melanotic masses in Drosophila larvae. Our study suggests that UBIAD1 serves as a tumor suppressor in cancer and tentatively reveals the underlying mechanism of melanotic mass formation in Drosophila.

Filamin A (FLNA) regulates autophagy of bladder carcinoma cell and affects its proliferation, invasion and metastasis.

This research intended to explore the effect of FLNA on cell proliferation, invasion and migration in bladder carcinoma (BC). Microarray analysis was performed with the TCGA data, and the results were confirmed on 20 paired BC tissues and adjacent tissues using qRT-PCR and immunohistochemistry. Transmission electron microscope (TEM) and cell fluorescence assay were used to observe the quantity of autophagosomes. The expression of autophagy-related protein (LC3-I/II, p62) was detected by western blot. Cell proliferation was detected using CCK-8 and colony formation. The invasion and migration ability of the cell were tested by transwell and wound-healing assay. Tumor xenograft in BALB/c nude mice and HE staining were utilized to probe into the effects of FLNA-induced regulation of volume, weight and metastasis of tumors. We confirmed that FLNA was down-regulated in BC tissues. TEM and fluorescence analysis proved that FLNA overexpression promoted autophagy in BC cells. Colony formation assay and CCK-8 experiment showed that FLNA overexpression suppressed the proliferation of BC cells. In addition, FLNA blocked cell cycle and promoted apoptosis of BC cell. Transwell assay and wound-healing assay further proved that FLNA suppressed invasion and migration ability in BC cell. Meanwhile, in vivo study indicated that FLNA inhibited the tumor growth. Overexpression of FLNA suppressed the proliferation, migration and invasion of the BC cell, suggesting the potential role of FLNA in clinical treatment.

CKAP4/p63 Is a Receptor for the Frizzled-8 Protein-related Antiproliferative Factor from Interstitial Cystitis Patients

Antiproliferative factor (APF) is a low molecular weight sialoglycopeptide that is secreted by bladder cells from interstitial cystitis patients and is a potent inhibitor of both normal bladder epithelial and bladder carcinoma cell proliferation. We hypothesized that APF may produce its antiproliferative effects by binding to a transmembrane receptor. This study demonstrates that cytoskeleton-associated protein 4/p63 (CKAP4/p63), a type II transmembrane receptor, binds with high affinity to APF. The antiproliferative activity of APF is effectively inhibited by preincubation with anti-CKAP4/p63-specific antibodies, as well as by short interfering RNA knockdown of CKAP4/p63. Immunofluorescent confocal microscopy showed co-localization of anti-CKAP4/p63 and rhodamine-labeled synthetic APF binding in both cell membrane and perinuclear areas. APF also inhibits the proliferation of HeLa cervical carcinoma cells that are known to express CKAP4/p63. These data indicate that CKAP4/p63 is an important epithelial cell receptor for APF.

DNA topoisomerase 억제제인 β-lapachone에 의한 인체 간암 및 방광암세포 증식억제에 관한 연구

남미지역에서 자생하는 Tabebuia avellanedae라는 나무의 수피에서 동정된 quinone계 물질이며, DNA topoisomeras억제제로 알려진 <TEX>$\beta-lapachone$</TEX>의 항암작용에 관한 부가적인 자료를 얻기 위하여 인체 간암(HepG2) 및 방광암(T24)세포를 대상으로 조사한 결과 다음과 같은 결과를 얻게 되었다. MTT assay 및 flow cytometry 분석 등의 결과에서, <TEX>$\beta-lapachone$</TEX>의 처리에 따라 조사된 두 가지 암세포에서 <TEX>$\beta-lapachone$</TEX>처리 농도의존적으로 암세포의 심한 형태적 변형이 동반되면서 암세포의 증식이 억제되었으며, 생존율이 저하되었고 이는 apoptosis유발과 상관성이 있음을 알 수 있었다. <TEX>$\beta-lapachone$</TEX>처리에 의한 두 암세포의 증식억제는 종양억제 유전자 p53 및 Cdk inhibitor p21의 발현과는 큰 연관성이 없음을 RT-PCR 및 Western blot analysis를 통하여 확인하였다. 그러나 전사조절인자 Sp-1 및 세포증식 주요조절인자인 PCNA의 단백질 발현은 <TEX>$\beta-lapachone$</TEX>처리에 따라 매우 감소되었으며, telomere조절에 중요한 인자들의 선택적 발현 저하 현상도 관찰되었다. 이상의 결과들은 인체 암세포에서 <TEX>$\beta-lapachone$</TEX>의 항암작용을 이해하는 중요한 자료가 될 것이며, <TEX>$\beta-lapachone$</TEX>과 유사한 화학적 구조 및 성질을 가지는 항암제 후보물질들의 항암기전 비교 및 항암제 개발을 위한 기초 자료로서 응용될 것이다. The objective of the present study was to investigate the effect of <TEX>$\beta-lapachone$</TEX>, a quinone obtained from the bark of the lapacho tree (Tabebuia avellanedae) in South America, on the cell growth of human hepatoma (HepG2) and bladder (T24) carcinoma cells. Exposure of cancer cells to <TEX>$\beta-lapachone$</TEX> resulted in growth inhibition, morphological changes and apoptosis in a concentration-dependent manner, which could be proved by MTT assay and flow cytometry analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses revealed that <TEX>$\beta-lapachone$</TEX> did not affect the levels of tumor suppressor p53 and cyclin-dependent kinase (Cdk) inhibitor p21 (WAFl/CIPl) expression. However, the transcriptional factor Sp-l and proliferating cell nuclear antigen (PCNA) protein levels were significantly down-regulated by <TEX>$\beta-lapachone$</TEX> in both cell lines. Moreover, <TEX>$\beta-lapachone$</TEX> treatment caused a dose-dependent inhibition of the expression of telomere regulatory gene products such as human telomere reverse transcriptase (hTERT) and telomerase-associated protein-l (TEP-l). Taken together, these findings suggest that <TEX>$\beta-lapachone$</TEX>-induced inhibition of human hepatoma and bladder carcinoma cell proliferation is associated with the induction of apoptotic cell death via modulation of several major growth regulatory gene products, and provide important new insights into the additional mechanisms of the anti-cancer activity of <TEX>$\beta-lapachone$</TEX>.

Methoxypsoralen phototherapy oftransitional cell carcinoma

Objectives. The goal of this research was to assess whether methoxypsoralencompounds in combination with ultraviolet light were effective in preventing cellular proliferation in an in vitro model of human transitional cell carcinoma. Methods. Three methoxypsoralen compounds, 5-methoxypsoralen (5-MOP), 8-methoxypsoralen (8-MOP), and 4′-aminomethyl 4,5′-8′-trimethylpsoralen (AMT), were added in vitro to T-24 transitional cell carcinoma cells. Psoralens directly bind to DNA, cross-linking the strands when exposed to ultraviolet light and thereby prevent cellular division. Results. In vitro activity was demonstrated utilizing AMT and ultraviolet radiation at 320 to 340 nm, preventing cellular proliferation in T-24 transitional cell carcinoma. Conclusions. Methoxypsoralen compounds in combination with ultraviolet light are effective in preventing proliferation of bladder carcinoma cells in vitro. This therapy may prove to be effective in clinical early stage transitional cell carcinoma and warrants further assessment.

The inhibitory effect of ciprofloxacin on proliferation of a murine bladder carcinoma cell line.

Ciprofloxacin was tested for its effect on proliferation of a bladder carcinoma cell line, Jurkat T-cell leukaemia cell line and a normal human foreskin derived from a fibroblast cell line. Cell proliferation was measured by MTT tetrazolium salt colorimetric assay. Proliferation of bladder carcinoma cells was inhibited by ciprofloxacin in a dose and time dependent fashion. Initial inhibition was observed at a concentration of 25 mg/L (approximately 35% inhibition; P greater than 0.01) while 250 mg/L of ciprofloxacin, a concentration achievable in urine following conventional oral treatment, exerted a lytic effect on these cells. A maximum suppressive effect on cell proliferation was reached within the first 24 h of ciprofloxacin addition to cell cultures. The rate of reversal of the inhibition of proliferation was dependent on the initial drug concentration. Exposure of bladder carcinoma cell cultures to 250 mg/L ciprofloxacin resulted in irreversible inhibition of proliferation. Similar results were obtained for the fibroblast cell line and Jurkat T-cell line. The latter being slightly more sensitive towards ciprofloxacin treatment in vitro.