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Finding ATF4/p75NTR/IL-8 signal pathway in endothelial-mesenchymal transition by safrole oxide.

Ge D, Jing Q, Zhao W, Yue H, Su L, Zhang S, Zhao J - PLoS ONE (2014)

Bottom Line: Safrole oxide (SFO), 50 µg/ml, could most effectively induce EndoMT within 12 h.Furthermore, knockdown of p75NTR inhibited the SFO-increased IL-8 expression and secretion, and knockdown of ATF4 inhibited SFO-increased p75NTR level significantly.The ATF4/p75NTR/IL-8 signal pathway may have an important role in EndoMT induced by SFO.

View Article: PubMed Central - PubMed

Affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Jinan, China.

ABSTRACT
Targeting the endothelial-to-mesenchymal transition (EndoMT) may be a novel therapeutic strategy for cancer and various diseases induced by fibrosis. We aimed to identify a small chemical molecule as an inducer of EndoMT and find a new signal pathway by using the inducer. Safrole oxide (SFO), 50 µg/ml, could most effectively induce EndoMT within 12 h. To understand the underlying molecular mechanism, we performed microarray, quantitative real-time PCR and western blot analysis to find key factors involved in SFO-induced EndoMT and demonstrated the involvement of the factors by RNAi. The expression of activating transcription factor 4 (ATF4), p75 neurotrophin receptor (p75NTR), and interleukin 8 (IL-8) was greatly increased in SFO-induced EndoMT. Knockdown of ATF4 inhibited the SFO-induced EndoMT completely, and knockdown of p75NTR or IL-8 partially inhibited the EndoMT, which suggests that all three factors were involved in the process. Furthermore, knockdown of p75NTR inhibited the SFO-increased IL-8 expression and secretion, and knockdown of ATF4 inhibited SFO-increased p75NTR level significantly. The ATF4/p75NTR/IL-8 signal pathway may have an important role in EndoMT induced by SFO. Our findings support potential novel targets for the therapeutics of cancer and fibrosis disease.

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Phase-contrast photomicrographs (A) and morphometric analyses (B) of human umbilical vascular endothelial cells (HUVECs).HUVECs were cultured in normal medium without or with 50 µg/ml safrole oxide (SFO) for 3, 6 and 12 h. (A) Cells in the control group retained cobblestone features, whereas cells cultured with SFO had elongated features and were arranged into “streamlined” structures. (B) Cell circularity, where a circle equals 0 and increasing elongation approaches 1, was calculated after SFO treatment (**p<0.01, n = 5).
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pone-0099378-g001: Phase-contrast photomicrographs (A) and morphometric analyses (B) of human umbilical vascular endothelial cells (HUVECs).HUVECs were cultured in normal medium without or with 50 µg/ml safrole oxide (SFO) for 3, 6 and 12 h. (A) Cells in the control group retained cobblestone features, whereas cells cultured with SFO had elongated features and were arranged into “streamlined” structures. (B) Cell circularity, where a circle equals 0 and increasing elongation approaches 1, was calculated after SFO treatment (**p<0.01, n = 5).

Mentions: Control HUVECs showed a cobblestone morphology. When HUVECs were exposed to a high concentration of SFO (50–70 µg/ml) for 3, 6 or 12 h, cells increasingly showed an elongated appearance. Furthermore, the cells were aligned, to create a “streamlined” effect with extended treatment [21] (Fig. 1A). Circularity analysis showed that HUVECs stimulated by 50 µg/ml SFO were more elongated, with circularity measurement ranging from 0.2 to 0.7 (p<0.01) (Fig. 1B). The phenomenon was sustained for 12 h. After then, smooth muscle-like cells undergo apoptosis [8].


Finding ATF4/p75NTR/IL-8 signal pathway in endothelial-mesenchymal transition by safrole oxide.

Ge D, Jing Q, Zhao W, Yue H, Su L, Zhang S, Zhao J - PLoS ONE (2014)

Phase-contrast photomicrographs (A) and morphometric analyses (B) of human umbilical vascular endothelial cells (HUVECs).HUVECs were cultured in normal medium without or with 50 µg/ml safrole oxide (SFO) for 3, 6 and 12 h. (A) Cells in the control group retained cobblestone features, whereas cells cultured with SFO had elongated features and were arranged into “streamlined” structures. (B) Cell circularity, where a circle equals 0 and increasing elongation approaches 1, was calculated after SFO treatment (**p<0.01, n = 5).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048316&req=5

pone-0099378-g001: Phase-contrast photomicrographs (A) and morphometric analyses (B) of human umbilical vascular endothelial cells (HUVECs).HUVECs were cultured in normal medium without or with 50 µg/ml safrole oxide (SFO) for 3, 6 and 12 h. (A) Cells in the control group retained cobblestone features, whereas cells cultured with SFO had elongated features and were arranged into “streamlined” structures. (B) Cell circularity, where a circle equals 0 and increasing elongation approaches 1, was calculated after SFO treatment (**p<0.01, n = 5).
Mentions: Control HUVECs showed a cobblestone morphology. When HUVECs were exposed to a high concentration of SFO (50–70 µg/ml) for 3, 6 or 12 h, cells increasingly showed an elongated appearance. Furthermore, the cells were aligned, to create a “streamlined” effect with extended treatment [21] (Fig. 1A). Circularity analysis showed that HUVECs stimulated by 50 µg/ml SFO were more elongated, with circularity measurement ranging from 0.2 to 0.7 (p<0.01) (Fig. 1B). The phenomenon was sustained for 12 h. After then, smooth muscle-like cells undergo apoptosis [8].

Bottom Line: Safrole oxide (SFO), 50 µg/ml, could most effectively induce EndoMT within 12 h.Furthermore, knockdown of p75NTR inhibited the SFO-increased IL-8 expression and secretion, and knockdown of ATF4 inhibited SFO-increased p75NTR level significantly.The ATF4/p75NTR/IL-8 signal pathway may have an important role in EndoMT induced by SFO.

View Article: PubMed Central - PubMed

Affiliation: Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Jinan, China.

ABSTRACT
Targeting the endothelial-to-mesenchymal transition (EndoMT) may be a novel therapeutic strategy for cancer and various diseases induced by fibrosis. We aimed to identify a small chemical molecule as an inducer of EndoMT and find a new signal pathway by using the inducer. Safrole oxide (SFO), 50 µg/ml, could most effectively induce EndoMT within 12 h. To understand the underlying molecular mechanism, we performed microarray, quantitative real-time PCR and western blot analysis to find key factors involved in SFO-induced EndoMT and demonstrated the involvement of the factors by RNAi. The expression of activating transcription factor 4 (ATF4), p75 neurotrophin receptor (p75NTR), and interleukin 8 (IL-8) was greatly increased in SFO-induced EndoMT. Knockdown of ATF4 inhibited the SFO-induced EndoMT completely, and knockdown of p75NTR or IL-8 partially inhibited the EndoMT, which suggests that all three factors were involved in the process. Furthermore, knockdown of p75NTR inhibited the SFO-increased IL-8 expression and secretion, and knockdown of ATF4 inhibited SFO-increased p75NTR level significantly. The ATF4/p75NTR/IL-8 signal pathway may have an important role in EndoMT induced by SFO. Our findings support potential novel targets for the therapeutics of cancer and fibrosis disease.

Show MeSH
Related in: MedlinePlus