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Constitutive and Inducible Expression of Invasion-related Factors in PC-3 Prostate Cancer Cells.

Hwang YS, Lindholm PF - J Cancer Prev (2015)

Bottom Line: Thrombospondin-1, interleukin-8, kallikrein 6, matrix metalloproteinase-1, and tissue factor were overexpressed in the highly invasive PC-3 variant cells and further upregulated by LPA stimulation.The results suggest that the target molecules are involved in invasiveness of prostate cancer.These molecules may have clinical value for anti-invasion therapy by serving as biomarkers for the prediction of aggressive cancers and the detection of pharmacological inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Department of Dental Hygiene, College of Health Science, Eulji University, Seongnam, Korea.

ABSTRACT

Background: Tumor growth and invasion are interconnected with the tumor microenvironment. Overexpression of genes that regulate cancer cell invasion by growth factors, cytokines, and lipid factors can affect cancer aggressiveness. A comparative gene expression analysis between highly invasive and low invasive cells revealed that various genes are differentially expressed in association with invasive potential. In this study, we selected variant PC-3 prostate cancer cell sublines and discovered critical molecules that contributed to their invasive potential.

Methods: The high invasive and low invasive variant PC-3 cell sublines were obtained by serial selection following Matrigel-coated Transwell invasion and were characterized by Transwell invasion, luciferase reporter assay, and Rhotekin pull-down assay. Lysophosphatidic acid (LPA) was added to the cultures to observe the response to this extracellular stimulus. The essential molecules related with cancer invasiveness were detected with Northern blotting, quantitative reverse transcription-polymerase chain reaction, and cDNA microarray.

Results: Highly invasive PC-3 cells showed higher nuclear factor kappa B (NF-κB), activator protein 1 (AP-1) and RhoA activities than of low invasive PC-3 cells. LPA promoted cancer invasion through NF-κB, AP-1, and RhoA activities. Thrombospondin-1, interleukin-8, kallikrein 6, matrix metalloproteinase-1, and tissue factor were overexpressed in the highly invasive PC-3 variant cells and further upregulated by LPA stimulation.

Conclusions: The results suggest that the target molecules are involved in invasiveness of prostate cancer. These molecules may have clinical value for anti-invasion therapy by serving as biomarkers for the prediction of aggressive cancers and the detection of pharmacological inhibitors.

No MeSH data available.


Related in: MedlinePlus

Lysophosphatidic acid (LPA)-stimulated expression of selected genes. Northern blot analysis was performed to determine the effect of LPA on the mRNA expression levels of several genes in the low invasive PC-3 prostate cancer cells. LPA was treated for 8 hours under serum-free condition. 18S and 28S are loading controls. MMP-1, matrix metalloproteinase-1.
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f3-jcp-20-121: Lysophosphatidic acid (LPA)-stimulated expression of selected genes. Northern blot analysis was performed to determine the effect of LPA on the mRNA expression levels of several genes in the low invasive PC-3 prostate cancer cells. LPA was treated for 8 hours under serum-free condition. 18S and 28S are loading controls. MMP-1, matrix metalloproteinase-1.

Mentions: Total RNA from the variant PC-3 cell sublines was examined by microarray analysis to verify changes in gene expression. As shown Table 2, the level of TSP-1, IL-8, KLK6, MMP-1, and TF were significantly different between the high and low invasive PC-3 cells. The PC-3 cells were treated with LPA and a Northern blot was performed to demonstrate whether expression of these genes could be induced by this stimulus. LPA induced the expression of these genes under serum-free conditions, although there were differences in the degree of response (Fig. 3). The same changes in gene expression were observed by the RT-PCR analysis with specific probes (Table 3), although the fold-change in expression was not always the same using these two different analytical methods. Expression of the differentially regulated genes in the microarray results with the variant PC-3 cell sublines were in direct agreement with the LPA-induced results.


Constitutive and Inducible Expression of Invasion-related Factors in PC-3 Prostate Cancer Cells.

Hwang YS, Lindholm PF - J Cancer Prev (2015)

Lysophosphatidic acid (LPA)-stimulated expression of selected genes. Northern blot analysis was performed to determine the effect of LPA on the mRNA expression levels of several genes in the low invasive PC-3 prostate cancer cells. LPA was treated for 8 hours under serum-free condition. 18S and 28S are loading controls. MMP-1, matrix metalloproteinase-1.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492356&req=5

f3-jcp-20-121: Lysophosphatidic acid (LPA)-stimulated expression of selected genes. Northern blot analysis was performed to determine the effect of LPA on the mRNA expression levels of several genes in the low invasive PC-3 prostate cancer cells. LPA was treated for 8 hours under serum-free condition. 18S and 28S are loading controls. MMP-1, matrix metalloproteinase-1.
Mentions: Total RNA from the variant PC-3 cell sublines was examined by microarray analysis to verify changes in gene expression. As shown Table 2, the level of TSP-1, IL-8, KLK6, MMP-1, and TF were significantly different between the high and low invasive PC-3 cells. The PC-3 cells were treated with LPA and a Northern blot was performed to demonstrate whether expression of these genes could be induced by this stimulus. LPA induced the expression of these genes under serum-free conditions, although there were differences in the degree of response (Fig. 3). The same changes in gene expression were observed by the RT-PCR analysis with specific probes (Table 3), although the fold-change in expression was not always the same using these two different analytical methods. Expression of the differentially regulated genes in the microarray results with the variant PC-3 cell sublines were in direct agreement with the LPA-induced results.

Bottom Line: Thrombospondin-1, interleukin-8, kallikrein 6, matrix metalloproteinase-1, and tissue factor were overexpressed in the highly invasive PC-3 variant cells and further upregulated by LPA stimulation.The results suggest that the target molecules are involved in invasiveness of prostate cancer.These molecules may have clinical value for anti-invasion therapy by serving as biomarkers for the prediction of aggressive cancers and the detection of pharmacological inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Department of Dental Hygiene, College of Health Science, Eulji University, Seongnam, Korea.

ABSTRACT

Background: Tumor growth and invasion are interconnected with the tumor microenvironment. Overexpression of genes that regulate cancer cell invasion by growth factors, cytokines, and lipid factors can affect cancer aggressiveness. A comparative gene expression analysis between highly invasive and low invasive cells revealed that various genes are differentially expressed in association with invasive potential. In this study, we selected variant PC-3 prostate cancer cell sublines and discovered critical molecules that contributed to their invasive potential.

Methods: The high invasive and low invasive variant PC-3 cell sublines were obtained by serial selection following Matrigel-coated Transwell invasion and were characterized by Transwell invasion, luciferase reporter assay, and Rhotekin pull-down assay. Lysophosphatidic acid (LPA) was added to the cultures to observe the response to this extracellular stimulus. The essential molecules related with cancer invasiveness were detected with Northern blotting, quantitative reverse transcription-polymerase chain reaction, and cDNA microarray.

Results: Highly invasive PC-3 cells showed higher nuclear factor kappa B (NF-κB), activator protein 1 (AP-1) and RhoA activities than of low invasive PC-3 cells. LPA promoted cancer invasion through NF-κB, AP-1, and RhoA activities. Thrombospondin-1, interleukin-8, kallikrein 6, matrix metalloproteinase-1, and tissue factor were overexpressed in the highly invasive PC-3 variant cells and further upregulated by LPA stimulation.

Conclusions: The results suggest that the target molecules are involved in invasiveness of prostate cancer. These molecules may have clinical value for anti-invasion therapy by serving as biomarkers for the prediction of aggressive cancers and the detection of pharmacological inhibitors.

No MeSH data available.


Related in: MedlinePlus