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NF-κB-mediated miR-124 suppresses metastasis of non-small-cell lung cancer by targeting MYO10.

Sun Y, Ai X, Shen S, Lu S - Oncotarget (2015)

Bottom Line: Over-expression of miR-124 robustly attenuated migration and metastatic ability of the aggressive cells.Knockdown of MYO10 inhibited cell migration, whereas forced MYO10 expression markedly rescued miR-124-mediated suppression of cell metastasis.Additionally, we found an activated NF-κB-centered inflammatory loop in the highly aggressive cells leading to down-regulation of miR-124.

View Article: PubMed Central - PubMed

Affiliation: Lung Tumor Clinical Medical Center, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, China.

ABSTRACT
Recently, dysregulation of microRNAs plays a critical role in cancer metastasis. Here, an in vivo selection approach was used to generate highly aggressive NSCLC sub-cell lines followed by comparing the microRNAs expression using microarrays. miR-124 was notably deregulated in both highly invasive sub-cell lines and node-positive NSCLC specimens. Over-expression of miR-124 robustly attenuated migration and metastatic ability of the aggressive cells. MYO10 was subsequently identified as a novel functional downstream target of miR-124, and was up-regulated in node-positive NSCLC tissues. Knockdown of MYO10 inhibited cell migration, whereas forced MYO10 expression markedly rescued miR-124-mediated suppression of cell metastasis. Additionally, we found an activated NF-κB-centered inflammatory loop in the highly aggressive cells leading to down-regulation of miR-124. These results suggest that NF-κB-regulated miR-124 targets MYO10, inhibits cell invasion and metastasis, and is down-regulated in node-positive NSCLC.

No MeSH data available.


Related in: MedlinePlus

An IL6-NF-κB inflammatory loop is activated in aggressive NSCLC cells(A) Cytokine array of conditioned media of parental H522 cells and aggressive H522M3 cells. (B) The intensity of each blot compared to control was determined by Kodak image analyzer. (C) The secretion of IL-5, IL-6, IL-8 and MCP-1 were examined by ELISA. (D) ELISA detecting the secretion of IL-6 and IL-8 in H522 cells after treated with NF-κB inhibitor (10 μM Bay11-7082), or combined inhibition of AKT (10 μM LY294002) and STAT3 (5 μM Stattic), recombinant IL-6 (rIL-6, 50 ng/ml) and recombinant IL-8 (rIL-8, 50 ng/ml). *p < 0.05 and ***p < 0.001. (E) Western blot analysis of the expression of pNF-κB, pAKT, pSTAT3 and pERK1/2 in H522 cells after stimulation with rIL-6 (50 ng/ml). (F) An IL6-NF-κB feedback loop was schematically illustrated.
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Figure 6: An IL6-NF-κB inflammatory loop is activated in aggressive NSCLC cells(A) Cytokine array of conditioned media of parental H522 cells and aggressive H522M3 cells. (B) The intensity of each blot compared to control was determined by Kodak image analyzer. (C) The secretion of IL-5, IL-6, IL-8 and MCP-1 were examined by ELISA. (D) ELISA detecting the secretion of IL-6 and IL-8 in H522 cells after treated with NF-κB inhibitor (10 μM Bay11-7082), or combined inhibition of AKT (10 μM LY294002) and STAT3 (5 μM Stattic), recombinant IL-6 (rIL-6, 50 ng/ml) and recombinant IL-8 (rIL-8, 50 ng/ml). *p < 0.05 and ***p < 0.001. (E) Western blot analysis of the expression of pNF-κB, pAKT, pSTAT3 and pERK1/2 in H522 cells after stimulation with rIL-6 (50 ng/ml). (F) An IL6-NF-κB feedback loop was schematically illustrated.

Mentions: Recent studies have demonstrated that a number of cytokines, including IL-6, IL-8 and MCP-1/CCL2, play a role in invasion and metastasis. Utilizing an antibody cytokine array, we examined the levels of cytokine expression in parental and aggressive H522M3 cells. An increase in IL-6, IL-8 and MCP-1 secretion was observed in aggressive H522M3 cells (Figure 6A), as assessed by densitometry of the cytokine blots (Figure 6B). Utilizing an ELISA, we further confirmed that secretion of these cytokines was increased by 2- to 5-fold in aggressive cells, compared to parental cells (Figure 6C). The significant elevated production of IL-6 and IL-8 was consistently observed in the two highly aggressive cells compared to parental cells with a prominent upregulation of MCP-1 in the H522M3 cells.


NF-κB-mediated miR-124 suppresses metastasis of non-small-cell lung cancer by targeting MYO10.

Sun Y, Ai X, Shen S, Lu S - Oncotarget (2015)

An IL6-NF-κB inflammatory loop is activated in aggressive NSCLC cells(A) Cytokine array of conditioned media of parental H522 cells and aggressive H522M3 cells. (B) The intensity of each blot compared to control was determined by Kodak image analyzer. (C) The secretion of IL-5, IL-6, IL-8 and MCP-1 were examined by ELISA. (D) ELISA detecting the secretion of IL-6 and IL-8 in H522 cells after treated with NF-κB inhibitor (10 μM Bay11-7082), or combined inhibition of AKT (10 μM LY294002) and STAT3 (5 μM Stattic), recombinant IL-6 (rIL-6, 50 ng/ml) and recombinant IL-8 (rIL-8, 50 ng/ml). *p < 0.05 and ***p < 0.001. (E) Western blot analysis of the expression of pNF-κB, pAKT, pSTAT3 and pERK1/2 in H522 cells after stimulation with rIL-6 (50 ng/ml). (F) An IL6-NF-κB feedback loop was schematically illustrated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480748&req=5

Figure 6: An IL6-NF-κB inflammatory loop is activated in aggressive NSCLC cells(A) Cytokine array of conditioned media of parental H522 cells and aggressive H522M3 cells. (B) The intensity of each blot compared to control was determined by Kodak image analyzer. (C) The secretion of IL-5, IL-6, IL-8 and MCP-1 were examined by ELISA. (D) ELISA detecting the secretion of IL-6 and IL-8 in H522 cells after treated with NF-κB inhibitor (10 μM Bay11-7082), or combined inhibition of AKT (10 μM LY294002) and STAT3 (5 μM Stattic), recombinant IL-6 (rIL-6, 50 ng/ml) and recombinant IL-8 (rIL-8, 50 ng/ml). *p < 0.05 and ***p < 0.001. (E) Western blot analysis of the expression of pNF-κB, pAKT, pSTAT3 and pERK1/2 in H522 cells after stimulation with rIL-6 (50 ng/ml). (F) An IL6-NF-κB feedback loop was schematically illustrated.
Mentions: Recent studies have demonstrated that a number of cytokines, including IL-6, IL-8 and MCP-1/CCL2, play a role in invasion and metastasis. Utilizing an antibody cytokine array, we examined the levels of cytokine expression in parental and aggressive H522M3 cells. An increase in IL-6, IL-8 and MCP-1 secretion was observed in aggressive H522M3 cells (Figure 6A), as assessed by densitometry of the cytokine blots (Figure 6B). Utilizing an ELISA, we further confirmed that secretion of these cytokines was increased by 2- to 5-fold in aggressive cells, compared to parental cells (Figure 6C). The significant elevated production of IL-6 and IL-8 was consistently observed in the two highly aggressive cells compared to parental cells with a prominent upregulation of MCP-1 in the H522M3 cells.

Bottom Line: Over-expression of miR-124 robustly attenuated migration and metastatic ability of the aggressive cells.Knockdown of MYO10 inhibited cell migration, whereas forced MYO10 expression markedly rescued miR-124-mediated suppression of cell metastasis.Additionally, we found an activated NF-κB-centered inflammatory loop in the highly aggressive cells leading to down-regulation of miR-124.

View Article: PubMed Central - PubMed

Affiliation: Lung Tumor Clinical Medical Center, Shanghai Chest Hospital, Shanghai Jiao Tong University, Shanghai 200030, China.

ABSTRACT
Recently, dysregulation of microRNAs plays a critical role in cancer metastasis. Here, an in vivo selection approach was used to generate highly aggressive NSCLC sub-cell lines followed by comparing the microRNAs expression using microarrays. miR-124 was notably deregulated in both highly invasive sub-cell lines and node-positive NSCLC specimens. Over-expression of miR-124 robustly attenuated migration and metastatic ability of the aggressive cells. MYO10 was subsequently identified as a novel functional downstream target of miR-124, and was up-regulated in node-positive NSCLC tissues. Knockdown of MYO10 inhibited cell migration, whereas forced MYO10 expression markedly rescued miR-124-mediated suppression of cell metastasis. Additionally, we found an activated NF-κB-centered inflammatory loop in the highly aggressive cells leading to down-regulation of miR-124. These results suggest that NF-κB-regulated miR-124 targets MYO10, inhibits cell invasion and metastasis, and is down-regulated in node-positive NSCLC.

No MeSH data available.


Related in: MedlinePlus