scholarly journals sFLT01 modulates invasion and metastasis in prostate cancer DU145 cells by inhibition of VEGF/GRP78/MMP2&9 axis

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Sepideh Taghizadeh ◽  
Zahra-Soheila Soheili ◽  
Mehdi Sadeghi ◽  
Shahram Samiei ◽  
Ehsan Ranaei Pirmardan ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Cancer Cell Line ◽  
Host Cells ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Pcr Analysis ◽  
Du145 Cells ◽  
And Migration

Abstract Background About 90% of cancer-related deaths are due to metastasis of cancer cells, and angiogenesis is a critical step in this process. sFLT01 is a novel fusion protein and a dual-targeting agent that neutralizes both VEGF and PlGF proangiogenic activities. GRP78 dual effect in tumor growth and angiogenesis could be activated under VEGF stimulation. The current study was designed to investigate the inhibitory impact of sFLT01 protein on VEGF/GRP78 axis. To this point, sFLT01 construct was synthesized, recombinant plasmid was expressed in eukaryotic host cells, sFLT01-HisTag protein was extracted and analyzed. The functional activity of sFLT01 on VEGF-enhanced tube formation and angiogenesis of HUVEC cells were examined. Eventually, the inhibitory impact of sFLT01 on growth, invasiveness, and migration of human prostate cancer cell line, DU145, was assessed. Real-time PCR evaluated the level of GRP78 and its effect on the downstream factors; matrix metallopeptidase proteins 2&9 (MMP2&9) along with tissue inhibitor of metalloproteinase proteins1&2 (TIMP1&2) under sFLT01 stimulation. Results According to the data, sFLT01 protein showed modulatory impact on proliferation, invasion, and migration of DU145 cells along with the potential of HUVECs angiogenesis. Real-Time PCR analysis depicted a significant downregulation in GRP78, MMP2 and MMP9 transcripts’ levels, and a subsequent elevation of TIMP1 and TIMP2 expression under sFLT01 stimulation was detected. Conclusion Overall, these data indicated that the inhibitory impact of sFLT01 on cancer cells growth and invasiveness could be mediated through the modulation of VEGF/GRP78/MMP2&9 axis and activation of TIMPs.

scholarly journals sFLT01 Modulates Invasion and Metastasis in Prostate Cancer DU145 Cells By Inhibition of VEGF/GRP78/MMP2&9 Axis

2021 ◽  
Author(s):  
Sepideh Taghizadeh ◽  
Zahra-Soheila Soheili ◽  
Mehdi Sadeghi ◽  
shahram Samiei ◽  
Fahimeh Zakeri ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Cancer Cell Line ◽  
Host Cells ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Pcr Analysis ◽  
Du145 Cells ◽  
And Migration

Abstract BackgroundAbout 90% of cancer-related deaths are due to metastasis of cancer cells and angiogenesis is a critical step in this process. sFLT01 is a novel fusion protein and a dual targeting agent that neutralizes both VEGF and PlGF proangiogenic activities. GRP78 dual effect in tumor growth and angiogenesis could be activated under VEGF stimulation. The current study was designed to investigate the inhibitory impact of sFLT01 protein on VEGF/GRP78 axis. To this point, sFLT01 construct was synthesized, recombinant plasmid was expressed in eukaryotic host cells, sFLT01-HisTag protein was extracted and analyzed. The functional activity of sFLT01 on VEGF-enhanced tube formation and angiogenesis of HUVEC cells were examined. Eventually, the inhibitory impact of sFLT01 on growth and invasiveness and migration of human prostate cancer cell line, DU145, was assessed. Real-time PCR evaluated the level of GRP78 and its effect on the downstream factors; matrix metallopeptidase proteins 2&9 (MMP2&9) along with tissue inhibitor of metalloproteinase proteins1&2 (TIMP1&2). Results According to the data, sFLT01 protein showed modulatory impact on proliferation, invasion, and migration of DU145 cells along with the potential of HUVECs angiogenesis. Real-Time PCR analysis depicted a significant downregulation in GRP78, MMP2 and MMP9 transcripts’ levels, and a subsequent elevation of TIMP1 and TIMP2 expression under sFLT01 stimulation was detected. Conclusion Overall, these data indicated that the inhibitory impact of sFLT01 on cancer cells growth and invasiveness could be mediated through the modulation of VEGF/GRP78/MMP2&9 axis and activation of TIMPs.

1435 POSTER Prostate Cancer Prognosis by Real-time PCR Analysis of PITX2 Methylation

2011 ◽  
Vol 47 ◽  
pp. S180
Author(s):  
O. Hasinger ◽  
P. Schatz ◽  
D. Dietrich ◽  
C. Ivascu ◽  
A. Sledziewski ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Real Time ◽  
Real Time Pcr ◽  
Cancer Prognosis ◽  
Pcr Analysis ◽  
Prostate Cancer Prognosis

The effects of L-NAME on DU145 human prostate cancer cell line: A cytotoxicity-based study

2019 ◽  
Vol 39 (2) ◽  
pp. 182-193
Author(s):  
S Kacar ◽  
F Kar ◽  
C Hacioglu ◽  
G Kanbak ◽  
V Sahinturk
Keyword(s):  
Oxidative Stress ◽  
Prostate Cancer ◽  
Antioxidant Capacity ◽  
Cancer Cells ◽  
Cancer Cell Line ◽  
Chemotherapeutic Agents ◽  
Human Prostate ◽  
Human Prostate Cancer ◽  
Bax Protein ◽  
Du145 Cells

Of all cancer types, prostate cancer is the second most common one with an age-standardized incidence rate of 29.3 per 100,000 men worldwide. Nitric oxide (NO) is both a radical and versatile messenger molecule involved in many physiological activities. NO was documented to be highly secreted and utilized by cancer cells. N ω-nitro-L-arginine methyl ester (L-NAME) is utilized for inhibiting NO synthase. Its worst long-term side effect is reported to be hypertension, hence less cytotoxic than chemotherapeutic agents. Herein, we carried out a cytotoxicity study on how different doses of L-NAME affect DU145 human prostate cancer cells. First, toxic doses of L-NAME were determined. Then, while antioxidant capacity was determined by glutathione and total antioxidant status, oxidative stress was evaluated by quantifying malondialdehyde, NO, and total oxidant status levels. Inflammatory effects of L-NAME were investigated by measuring tumor necrosis factor- α and interleukin-6 (IL-6) levels. Apoptotic effects of L-NAME were evaluated by measuring cytochrome C somatic and caspase 3 levels and by staining Bax protein. Finally, morphological analysis was performed. IC50 of L-NAME against DU145 cells was 12.2 mM. In L-NAME-treated DU145 cells, a dose-dependent increase in oxidative stress, inflammatory, and apoptotic marker proteins and decrease in antioxidant capacity were observed. While at the moderate dose of L-NAME, apoptotic changes were commonly observed, at higher doses, vacuolated and swollen cells were also recorded. We believe that the present study will encourage future studies by providing insights about dose and effects of L-NAME.

scholarly journals β2AR-HIF-1α-CXCL12 signaling of osteoblasts activated by isoproterenol promotes migration and invasion of prostate cancer cells

2019 ◽  
Author(s):  
Zhibin Huang ◽  
Guihuan Li ◽  
Zhishuai Zhang ◽  
Ruonan Gu ◽  
Wenyang Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Bone Metastasis ◽  
Cancer Cells ◽  
Chronic Stress ◽  
Adrenergic Receptor ◽  
Migration And Invasion ◽  
Prostate Cancer Cells ◽  
Mesenchymal Transition ◽  
Du145 Cells ◽  
And Migration

Abstract Background: Chronic stress is well known to promote tumor progression, however, little is known on whether neurotransmitters released after chronic stress induced sympathetic activation regulate the function of osteoblasts to affect migration and invasion of metastatic cancer cells. Methods: First, the changes in migration and invasion ability of prostate cancer cell lines PC-3 and DU145 were assessed by transwell migration assay. PC-3 and DU145 cells proliferation ability were detected by CCK-8, and HIF-1α expression of osteoblasts and the momentous proteins of epithelial-mesenchymal transition (EMT) of PC-3 and DU145 cells were examined by Western blot. Then, an analysis of the main cytokines associated with bone metastasis in osteoblasts and EMT-related biomarkers in PC-3 and DU145 cells was performed by qRT-PCR. Finally, rescue experiment was performed by using HIF-1α siRNA and inhibitor, HIF-1α overexpression plasmid, β2-adrenergic receptor (β2AR) inhibitor, CXCR4 siRNA and inhibitor. Results: In this study, isoproterenol (ISO), a non-selective β-adrenergic receptor (βAR) agonist, used as a pharmacological surrogate of sympathetic nerve activation induced by chronic stress, exhibited no direct effect on migration and invasion of PC-3 and DU145 prostate cancer cells. Whereas, osteoblasts pretreated with ISO promoted EMT and migration as well as invasion of PC-3 and DU145 cells, which was independent of promoting cell proliferation and could be inhibited by β2AR inhibitor. Mechanically, ISO increased the secretion of CXCL12 via the β2AR-HIF-1α signaling in osteoblasts. Moreover, overexpression of HIF-1α osteoblasts promoted migration and invasion of PC-3 and DU145 cells, which was inhibited by addition of recombinant knockdown of CXCR4 in PC-3 and DU145 cells, and inhibiting CXCL12-CXCR4 signaling with LY2510924 blunted the effects of osteoblasts in response to ISO on EMT and migration as well as invasion of PC-3 and DU145 cells. Conclusions: These findings indicate that β2AR-HIF-1α-CXCL12 signaling in osteoblasts facilitates migration and invasion as well as EMT of prostate cancer cells, and may play a potential role in affecting bone metastasis of prostate cancer.

scholarly journals β2AR-HIF-1α-CXCL12 signaling of osteoblasts activated by isoproterenol promotes migration and invasion of prostate cancer cells

2019 ◽  
Author(s):  
Zhibin Huang ◽  
Guihuan Li ◽  
Zhishuai Zhang ◽  
Ruonan Gu ◽  
Wenyang Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Bone Metastasis ◽  
Cancer Cells ◽  
Chronic Stress ◽  
Adrenergic Receptor ◽  
Migration And Invasion ◽  
Prostate Cancer Cells ◽  
Mesenchymal Transition ◽  
Du145 Cells ◽  
And Migration

Abstract Background: Chronic stress is well known to promote tumor progression, however, little is known on whether neurotransmitters released after chronic stress induced sympathetic activation regulate the function of osteoblasts to affect migration and invasion of metastatic cancer cells. Methods: First, the changes in migration and invasion ability of prostate cancer cell lines PC-3 and DU145 were assessed by transwell migration assay. PC-3 and DU145 cells proliferation ability were detected by CCK-8, and HIF-1α expression of osteoblasts and the momentous proteins of epithelial-mesenchymal transition (EMT) of PC-3 and DU145 cells were examined by Western blot. Then, an analysis of the main cytokines associated with bone metastasis in osteoblasts and EMT-related biomarkers in PC-3 and DU145 cells was performed by qRT-PCR. Finally, rescue experiment was performed by using HIF-1α siRNA and inhibitor, HIF-1α overexpression plasmid, β2-adrenergic receptor (β2AR) inhibitor, CXCR4 siRNA and inhibitor. Results: In this study, isoproterenol (ISO), a non-selective β-adrenergic receptor (βAR) agonist, used as a pharmacological surrogate of sympathetic nerve activation induced by chronic stress, exhibited no direct effect on migration and invasion of PC-3 and DU145 prostate cancer cells. Whereas, osteoblasts pretreated with ISO promoted EMT and migration as well as invasion of PC-3 and DU145 cells, which was independent of promoting cell proliferation and could be inhibited by β2AR inhibitor. Mechanistically, ISO increased the secretion of CXCL12 via the β2AR-HIF-1α signaling in osteoblasts. Moreover, overexpression of HIF-1α osteoblasts promoted migration and invasion of PC-3 and DU145 cells, which was inhibited by addition of recombinant knockdown of CXCR4 in PC-3 and DU145 cells, and inhibiting CXCL12-CXCR4 signaling with LY2510924 blunted the effects of osteoblasts in response to ISO on EMT and migration as well as invasion of PC-3 and DU145 cells. Conclusions: These findings indicate that β2AR-HIF-1α-CXCL12 signaling in osteoblasts facilitates migration and invasion as well as EMT of prostate cancer cells, and may play a potential role in affecting bone metastasis of prostate cancer.

scholarly journals β2AR-HIF-1α-CXCL12 signaling of osteoblasts activated by isoproterenol promotes migration and invasion of prostate cancer cells

2019 ◽  
Author(s):  
Zhibin Huang ◽  
Guihuan Li ◽  
Zhishuai Zhang ◽  
Ruonan Gu ◽  
Wenyang Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Bone Metastasis ◽  
Cancer Cells ◽  
Chronic Stress ◽  
Adrenergic Receptor ◽  
Migration And Invasion ◽  
Prostate Cancer Cells ◽  
Mesenchymal Transition ◽  
Du145 Cells ◽  
And Migration

Abstract Background Chronic stress is well known to promote tumor progression, however, the mechanisms that underlie the association of chronic stress with cancer metastasis remain elusive. Methods First, the changes in migration and invasion ability of prostate cancer cell lines PC-3 and DU145 were assessed by transwell migration assay. And HIF-1α expression of osteoblasts and the momentous proteins of epithelial-mesenchymal transition (EMT) of PC-3 and DU145 cells were examined by western blot. Then, an analysis of the main cytokines associated with bone metastasis was performed in osteoblasts by qRT-PCR. Finally, HIF-1α siRNA and inhibitor YC-1 were used to assess the reverse of isoproterenol (ISO)-induced changes of HIF-1α in osteoblasts, and β2-adrenergic receptor (β2AR) inhibitor ICI118,551 and CXCR4 inhibitor LY2510924 were used to antagonizes migration and invasion of PC-3 and DU145 cells induced by osteoblasts triggered by ISO. Results In this study, ISO, a non-selective β-adrenergic receptor (βAR) agonist, used as a pharmacological surrogate of sympathetic nerve activation induced by chronic stress, exhibits no direct effect on migration and invasion of PC-3 and DU145 prostate cancer cells. Whereas, osteoblasts pretreated with ISO promote EMT and migration as well as invasion of PC-3 and DU145 cells, which can be inhibited by β2AR inhibitor. Mechanically, ISO increases the secretion of CXCL12 via the β2AR-HIF-1α signaling in osteoblasts. Moreover, inhibiting CXCL12-CXCR4 signaling with LY2510924 blunts the effects of osteoblasts in response to ISO on EMT and migration as well as invasion of PC-3 and DU145 cells. Conclusions These findings indicate that β2AR-HIF-1α-CXCL12 signaling in osteoblasts facilitates migration and invasion as well as EMT of prostate cancer cells, and may play a potential role in affecting bone metastasis of prostate cancer.

scholarly journals Cytostatic Action of Novel Histone Deacetylase Inhibitors in Androgen Receptor-Null Prostate Cancer Cells

2021 ◽  
Vol 14 (2) ◽  
pp. 103
Author(s):  
Zohaib Rana ◽  
Joel D. A. Tyndall ◽  
Muhammad Hanif ◽  
Christian G. Hartinger ◽  
Rhonda J. Rosengren
Keyword(s):  
Prostate Cancer ◽  
Androgen Receptor ◽  
Cancer Cells ◽  
Hdac Inhibitors ◽  
G1 Arrest ◽  
Prostate Cancer Cells ◽  
Prostate Tumors ◽  
Normal Prostate ◽  
Pc3 Cells ◽  
Du145 Cells

Androgen receptor (AR)-null prostate tumors have been observed in 11–24% of patients. Histone deacetylases (HDACs) are overexpressed in prostate tumors. Therefore, HDAC inhibitors (Jazz90 and Jazz167) were examined in AR-null prostate cancer cell lines (PC3 and DU145). Both Jazz90 and Jazz167 inhibited the growth of PC3 and DU145 cells. Jazz90 and Jazz167 were more active in PC3 cells and DU145 cells in comparison to normal prostate cells (PNT1A) and showed a 2.45- and 1.30-fold selectivity and higher cytotoxicity toward DU145 cells, respectively. Jazz90 and Jazz167 reduced HDAC activity by ~60% at 50 nM in PC3 lysates. At 4 μM, Jazz90 and Jazz167 increased acetylation in PC3 cells by 6- to 8-fold. Flow cytometry studies on the cell phase distribution demonstrated that Jazz90 causes a G0/G1 arrest in AR-null cells, whereas Jazz167 leads to a G0/G1 arrest in DU145 cells. However, apoptosis only occurred at a maximum of 7% of the total cell population following compound treatments in PC3 and DU145 cells. There was a reduction in cyclin D1 and no significant changes in bcl-2 in DU145 and PC3 cells. Overall, the results showed that Jazz90 and Jazz167 function as cytostatic HDAC inhibitors in AR-null prostate cancer cells.

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