Analysis of tumor necrosis factor α-induced and nuclear factor κB-silenced LNCaP prostate cancer cells by RT-qPCR

Gonen-Korkmaz, Ceren; Sevin, Gülnur; Gökçe, Göksel; Arun, Mehmet Zuhuri; Yildirim, Gökçe; Reel, Buket; Kaymak, Ayşegül Öztürk; Ogut, Deniz

doi:10.3892/etm.2014.2032

Cited by 8 publications

(15 citation statements)

References 25 publications

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“…STAMP2 mRNA expression was significantly induced by all three cytokines in a dose-dependent manner (Figure 2a-c). Contrary to a previous report of negative results [35], TNF treatment of LNCaP cells led to a modest, but significant induction of STAMP2, although weaker if compared to the TNF-induced canonical NF-κB target gene IKBA (inhibitor of kappa B alpha) (Figure 2a). At the same concentration, TNF did not induce STAMP2 expression in C4-2B cells (Figure S1a).…”

Section: Temporal Regulation Of Stamp2 Expression By Cytokinescontrasting

confidence: 99%

STAMP2 Expression Mediated by Cytokines Attenuates Their Growth-Limiting Effects in Prostate Cancer Cells

Pihlstrøm

Jin

Nenseth

et al. 2021

Cancers

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Inflammatory events and dysregulated cytokine expression are implicated in prostate cancer (PCa), but the underlying molecular mechanisms are poorly understood at present. We have previously identified six transmembrane protein of the prostate 2 (STAMP2, also known as STEAP4) as an androgen-regulated gene, as well as a key regulator of PCa growth and survival. STAMP2 is also regulated by, and participates in, inflammatory signaling in other tissues and pathologies. Here, we show that the proinflammatory cytokines interleukin 6 (IL-6) and Interleukin 1 beta (IL-1β) significantly increase and strongly synergize in promoting STAMP2 expression in PCa cells. The two cytokines increase androgen-induced STAMP2 expression, but not expression of other known androgen target genes, suggesting a unique interplay of androgens and cytokines in regulating STAMP2 expression. Interestingly, STAMP2 knockdown significantly increased the ability of IL-6 and IL-1β to inhibit PCa cell growth in vitro. These results suggest that STAMP2 may represent a unique node through which inflammatory events mediate their effects on PCa growth and survival.

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Section: Temporal Regulation Of Stamp2 Expression By Cytokinescontrasting

confidence: 99%

STAMP2 Expression Mediated by Cytokines Attenuates Their Growth-Limiting Effects in Prostate Cancer Cells

Pihlstrøm

Jin

Nenseth

et al. 2021

Cancers

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“…This expression has then been found to increase when the prostate is in a diseased state (Korkmaz et al, 2002;Porkka et al, 2002;Wang et al, 2010). Previous studies have found STEAP2 to be most highly expressed in LNCaP cells, with lower but still increased levels recorded in PC3 cells, which correlate with the results of this study (Figure 3.1; Gonen- Korkmaz et al, 2014;Porkka et al, 2002;Whiteland et al, 2014). Of the four prostate cancer cell lines assessed here, DU145 cells were the only cell line to exhibit lower levels of STEAP2 than the normal cell line PNT2, consistent with previous studies (Figure 3.1; Korkmaz et al, 2002;Porkka et al, 2002).…”

Section: Challitasupporting

confidence: 90%

“…In previous studies, STEAP2 overexpression has been induced either ectopically or by transfection in DU145 cells to monitor its effects in androgenindependent cells, further suggesting its expression in this cell line is relatively low (Gonen- Korkmaz et al, 2014;Wang et al, 2010). The highest significant increases in STEAP2 expression were found in androgen-sensitive prostate cancer cell lines; LNCaP and C4-2B, suggesting that AR signalling may play a role in its expression, hence the inclusion of the androgen-sensitive, LNCaPderived C4-2B cell line in this study.…”

Section: Challitamentioning

confidence: 99%

The role of STEAP2 in aggressive prostate cancer traits and androgen response

Leia¹

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The prognosis of localised prostate cancer is generally promising, as many tumours remain dormant and therefore do not require immediate intervention. In contrast, once metastasised, the prognosis for aggressive prostate cancer is often poor, highlighting the need for novel, effective treatment approaches. The expression of the six transmembrane epithelial antigen of the prostate2 (STEAP2) cell surface protein is increased in aggressive prostate cancer compared to normal prostate tissue. In vitro studies have shown STEAP2 to aid in prostate cancer progression, and as such this molecule shows promise as a potential novel therapeutic target in the treatment of advanced disease. The aim of this thesis was to develop a comprehensive understanding of the mechanistic role of STEAP2 in promoting aggressive prostate cancer traits and evaluate if its knock-out has the capacity to reduce the invasive potential of prostate cancer cells in vitro. As prostate cancer is a largely androgen dependent disease, this thesis also aimed to evaluate the effects of STEAP2 inhibition on the expression of the androgen receptor and androgen-regulated genes. This study developed and optimised a protocol for generating a set of 3D prostate cancer spheroids to provide more representative models of the in vivo prostate cancer environment. In this thesis, one commercial anti-STEAP2 polyclonal antibody and a panel of anti-STEAP2 monoclonal antibodies were selected for proof-of-concept studies where their effects on reducing prostate cancer cell viability were assessed. Receptor internalisation of STEAP2 was evaluated upon anti-STEAP2 monoclonal antibody binding to determine its suitability for use with antibody-drug conjugate technology. STEAP2 expression was knocked out using CRISPR/Cas9 genome engineering technology in two prostate cancer cell lines to evaluate its impact on cell proliferation, migration and invasion. Furthermore, gene expression profiling was conducted to explore interactions between STEAP2, the androgen receptor and a panel of androgen-regulated genes (PSA, FKBP5, GPRC6A and TMPRSS2) following: 1) anti-STEAP2 antibody treatment, 2) STEAP2-knockout and 3) the growth of prostate cancer cells in androgen-depleted conditions. The data presented in this thesis demonstrate that inhibition of STEAP2 by both the polyclonal anti-STEAP2 antibody and lead anti-STEAP2 monoclonal antibody significantly reduced prostate cancer cell viability. STEAP2 receptor internalisation was triggered following treatment of prostate cancer cells with the anti-STEAP2 monoclonal antibody, demonstrating its potential utility with antibody-drug conjugate technology in the future. STEAP2 knockout prostate cancer cells exhibited decreased cell proliferation, migration and invasion in comparison to wild-type cells. These promising findings highlight the therapeutic value of STEAP2-knockout in inhibiting invasive tumour cell traits. Gene expression data from both STEAP2-knockout cells and androgen-depleted cells suggest that STEAP2 may be involved in crosstalk between the androgen receptor and androgen-regulated genes. STEAP2 could therefore provide a novel target in conjunction with current conventional androgen deprivation therapy. In conclusion, the in vitro findings presented herein suggest STEAP2 as a viable target for the development of more tailored and personalised therapeutic agents to improve the clinical management of men with aggressive prostate cancer.

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“…Western blot assays were performed according to previously described methods ( 10 , 11 , 13 ). Briefly, cMSCs were lysed with RIPA buffer containing phenylmethylsulfonyl fluoride (Sangon Biotech Co., Ltd.), then 50 µg total protein was quantitated using a BCA protein assay kit (P0009; Beyotime Institute of Biotechnology) and subjected to 6–10% SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

Electric pulse current stimulation increases electrophysiological properties of If current reconstructed in mHCN4-transfected canine mesenchymal stem cells

Yang

Luo

Pan

et al. 2016

Experimental and Therapeutic Medicine

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The ‘funny’ current, also known as the If current, play a crucial role in the spontaneous diastolic depolarization of sinoatrial node cells. The If current is primarily induced by the protein encoded by the hyperpolarization-activated cyclic nucleotide-gated channel 4 (HCN4) gene. The functional If channel can be reconstructed in canine mesenchymal stem cells (cMSCs) transfected with mouse HCN4 (mHCN4). Biomimetic studies have shown that electric pulse current stimulation (EPCS) can promote cardiogenesis in cMSCs. However, whether EPCS is able to influence the properties of the If current reconstructed in mHCN4-transfected cMSCs remains unclear. The present study aimed to investigate the effects of EPCS on the If current reconstructed in mHCN4-transfected cMSCs. The cMSCs were transfected with the lentiviral vector pLentis-mHCN4-GFP. Following transfection, these cells were divided into two groups: mHCN4-transfected cMSCs (group A), and mHCN4-transfected cMSCs induced by EPCS (group B). Using a whole cell patch-clamp technique, the If current was recorded, and group A cMSCs showed significant time and voltage dependencies and sensitivity to extracellular Cs+. The half-maximal activation (V1/2) value was −101.2±4.6 mV and the time constant of activation was 324±41 msec under −160 mV. In the group B cells the If current increased obviously and activation curve moved to right. The absolute value of V1/2 increased significantly to −92.4±4.8 mV (P<0.05), and the time constant of activation diminished under the same command voltage (251±44 vs. 324±41, P<0.05). In addition, the mRNA and protein expression levels of HCN4, connexin 43 (Cx43) and Cx45 were upregulated in group B compared with group A, as determined by reverse transcription-quantitative polymerase chain reaction and western blot analyses. Transmission electron micrographs also confirmed the increased gap junctions in group B. Collectively, these results indicated that reconstructed If channels may have a positive regulatory role in EPCS induction. The cMSCs transfected with mHCN4 induced by EPCS may be a source of effective biological pacemaker cells.

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Analysis of tumor necrosis factor α-induced and nuclear factor κB-silenced LNCaP prostate cancer cells by RT-qPCR

Cited by 8 publications

References 25 publications

STAMP2 Expression Mediated by Cytokines Attenuates Their Growth-Limiting Effects in Prostate Cancer Cells

STAMP2 Expression Mediated by Cytokines Attenuates Their Growth-Limiting Effects in Prostate Cancer Cells

The role of STEAP2 in aggressive prostate cancer traits and androgen response

Electric pulse current stimulation increases electrophysiological properties of If current reconstructed in mHCN4-transfected canine mesenchymal stem cells

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