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Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings promotes proliferation and survival of non-small cell lung cancer cells.

Li J, Lan T, Zhang C, Zeng C, Hou J, Yang Z, Zhang M, Liu J, Liu B - Oncotarget (2015)

Bottom Line: Inflammatory cytokines and oxidative stress are two critical mediators in inflammation-associated cancer.Interleukin-6 (IL-6) is one of the most critical tumor-promoting cytokines in non-small cell lung cancer (NSCLC).The in vivo results were similar to those obtained in vitro.

View Article: PubMed Central - PubMed

Affiliation: Clinical Pharmacy Department, Guangdong Pharmaceutical University, Guangzhou 510006, China.

ABSTRACT
Inflammatory cytokines and oxidative stress are two critical mediators in inflammation-associated cancer. Interleukin-6 (IL-6) is one of the most critical tumor-promoting cytokines in non-small cell lung cancer (NSCLC). In our recent study, we confirmed that NADPH oxidase 4 (NOX4), an important source of reactive oxygen species (ROS) production in NSCLC cells, promotes malignant progression of NSCLC. However, whether the crosstalk of NOX4 and IL-6 signalings exists in NSCLC remains undentified. In this study, we show that NOX4 expression is positively correlated with IL-6 expression in NSCLC tissues. Exogenous IL-6 treatment significantly enhances NOX4/ROS/Akt signaling in NSCLC cells. NOX4 also enhances IL-6 production and activates IL-6/STAT3 signaling in NSCLC cells. Specifically, NOX4 is confirmed to functionally interplay with IL-6 to promote NSCLC cell proliferation and survival. The in vivo results were similar to those obtained in vitro. These data indicate a novel NOX4-dependent link among IL-6 in the NSCLC microenvironment, oxidative stress in NSCLC cells and autocrined IL-6 in NSCLC cells. NOX4/Akt and IL-6/STAT3 signalings can reciprocally and positively regulate each other, leading to enhanced NSCLC cell proliferation and survival. Therefore, NOX4 may serve as a promising target against NSCLC alone with IL-6 signaling.

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Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings in H460 cells(A-B) The effects of IL-6 administration on NOX4 expression, Akt activity and ROS production in H460 cells at the indicated times, respectively. Bars are mean ± SD from four independent experiments. *Significantly different from control, P < 0.05. (C) H460 cells were stably transfected with control vector, NOX4 plasmid, respectively. Overexpression of NOX4 was confirmed by western blotting. (D-F) The effects of NOX4 overexpression on ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from four independent experiments. *Significantly different from vector control, P < 0.05. (G) H460 cells were stably transfected with scramble shRNA, NOX4 shRNA, respectively. Knockdown of NOX4 was analyzed by western blotting. (H-J) Silencing NOX4 significantly inhibited ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from five independent experiments. *Significantly different from vector control, P < 0.05.
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Figure 6: Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings in H460 cells(A-B) The effects of IL-6 administration on NOX4 expression, Akt activity and ROS production in H460 cells at the indicated times, respectively. Bars are mean ± SD from four independent experiments. *Significantly different from control, P < 0.05. (C) H460 cells were stably transfected with control vector, NOX4 plasmid, respectively. Overexpression of NOX4 was confirmed by western blotting. (D-F) The effects of NOX4 overexpression on ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from four independent experiments. *Significantly different from vector control, P < 0.05. (G) H460 cells were stably transfected with scramble shRNA, NOX4 shRNA, respectively. Knockdown of NOX4 was analyzed by western blotting. (H-J) Silencing NOX4 significantly inhibited ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from five independent experiments. *Significantly different from vector control, P < 0.05.

Mentions: To exclude the possibility that the observed effects are restricted to A549 cells, we sought to confirm the above effects in another NSCLC cell line H460. Like in A549 cells, IL-6 (10 ng/mL) treatment led to time-dependent increase in NOX4 level (Fig. 6A), Akt activity (Fig. 6A) as well as ROS production (Fig. 6B) in H460 cells. We also found that NOX4 overexpression (Fig. 6C) could significantly enhance ROS production (Fig. 6D), IL-6 expression (Fig. 6E) as well as the activity of its downstream JAK1 and STAT3 (Fig. 6F), while depletion of NOX4 (Fig. 6G) resulted in reduced ROS production (Fig. 6H), IL-6 expression (Fig. 6I) as well as JAK1 and STAT3 activity (Fig. 6J) in H460 cells.


Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings promotes proliferation and survival of non-small cell lung cancer cells.

Li J, Lan T, Zhang C, Zeng C, Hou J, Yang Z, Zhang M, Liu J, Liu B - Oncotarget (2015)

Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings in H460 cells(A-B) The effects of IL-6 administration on NOX4 expression, Akt activity and ROS production in H460 cells at the indicated times, respectively. Bars are mean ± SD from four independent experiments. *Significantly different from control, P < 0.05. (C) H460 cells were stably transfected with control vector, NOX4 plasmid, respectively. Overexpression of NOX4 was confirmed by western blotting. (D-F) The effects of NOX4 overexpression on ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from four independent experiments. *Significantly different from vector control, P < 0.05. (G) H460 cells were stably transfected with scramble shRNA, NOX4 shRNA, respectively. Knockdown of NOX4 was analyzed by western blotting. (H-J) Silencing NOX4 significantly inhibited ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from five independent experiments. *Significantly different from vector control, P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4359215&req=5

Figure 6: Reciprocal activation between IL-6/STAT3 and NOX4/Akt signalings in H460 cells(A-B) The effects of IL-6 administration on NOX4 expression, Akt activity and ROS production in H460 cells at the indicated times, respectively. Bars are mean ± SD from four independent experiments. *Significantly different from control, P < 0.05. (C) H460 cells were stably transfected with control vector, NOX4 plasmid, respectively. Overexpression of NOX4 was confirmed by western blotting. (D-F) The effects of NOX4 overexpression on ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from four independent experiments. *Significantly different from vector control, P < 0.05. (G) H460 cells were stably transfected with scramble shRNA, NOX4 shRNA, respectively. Knockdown of NOX4 was analyzed by western blotting. (H-J) Silencing NOX4 significantly inhibited ROS production, IL-6 production and JAK1/STAT3 activity after 24 hours. Bars are mean ± SD from five independent experiments. *Significantly different from vector control, P < 0.05.
Mentions: To exclude the possibility that the observed effects are restricted to A549 cells, we sought to confirm the above effects in another NSCLC cell line H460. Like in A549 cells, IL-6 (10 ng/mL) treatment led to time-dependent increase in NOX4 level (Fig. 6A), Akt activity (Fig. 6A) as well as ROS production (Fig. 6B) in H460 cells. We also found that NOX4 overexpression (Fig. 6C) could significantly enhance ROS production (Fig. 6D), IL-6 expression (Fig. 6E) as well as the activity of its downstream JAK1 and STAT3 (Fig. 6F), while depletion of NOX4 (Fig. 6G) resulted in reduced ROS production (Fig. 6H), IL-6 expression (Fig. 6I) as well as JAK1 and STAT3 activity (Fig. 6J) in H460 cells.

Bottom Line: Inflammatory cytokines and oxidative stress are two critical mediators in inflammation-associated cancer.Interleukin-6 (IL-6) is one of the most critical tumor-promoting cytokines in non-small cell lung cancer (NSCLC).The in vivo results were similar to those obtained in vitro.

View Article: PubMed Central - PubMed

Affiliation: Clinical Pharmacy Department, Guangdong Pharmaceutical University, Guangzhou 510006, China.

ABSTRACT
Inflammatory cytokines and oxidative stress are two critical mediators in inflammation-associated cancer. Interleukin-6 (IL-6) is one of the most critical tumor-promoting cytokines in non-small cell lung cancer (NSCLC). In our recent study, we confirmed that NADPH oxidase 4 (NOX4), an important source of reactive oxygen species (ROS) production in NSCLC cells, promotes malignant progression of NSCLC. However, whether the crosstalk of NOX4 and IL-6 signalings exists in NSCLC remains undentified. In this study, we show that NOX4 expression is positively correlated with IL-6 expression in NSCLC tissues. Exogenous IL-6 treatment significantly enhances NOX4/ROS/Akt signaling in NSCLC cells. NOX4 also enhances IL-6 production and activates IL-6/STAT3 signaling in NSCLC cells. Specifically, NOX4 is confirmed to functionally interplay with IL-6 to promote NSCLC cell proliferation and survival. The in vivo results were similar to those obtained in vitro. These data indicate a novel NOX4-dependent link among IL-6 in the NSCLC microenvironment, oxidative stress in NSCLC cells and autocrined IL-6 in NSCLC cells. NOX4/Akt and IL-6/STAT3 signalings can reciprocally and positively regulate each other, leading to enhanced NSCLC cell proliferation and survival. Therefore, NOX4 may serve as a promising target against NSCLC alone with IL-6 signaling.

Show MeSH
Related in: MedlinePlus