Low-Temperature Plasma-Activated Medium Inhibited Proliferation and Progression of Lung Cancer by Targeting the PI3K/Akt and MAPK Pathways

Figure 7

Potential pathways involved in PAM inhibition of lung tumor growth. (a) Volcano plot indicating 4319 significantly upregulated and 4877 significantly downregulated genes (; |log2 fold ) in the PAM-treated group compared with the control group. (b) KEGG analysis representing signaling pathways deferentially represented in control and PAM-treated 3D A549 spheroids. (c) Left: Western blot of phosphorylated PI3K and Akt in control and PAM-treated 3D A549 spheroids. Total protein levels of PI3K and Akt were analyzed. β-Actin was used as a loading control. Right: quantification of phosphorylated PI3K and Akt normalized to the total amount of the same proteins. (d) Left: Western blot of phosphorylated ERK, p38, and JNK in control and PAM-treated 3D A549 cells. Total protein levels of ERK, p38, and JNK were analyzed. β-Actin was used as a loading control. Right: quantification of phosphorylated ERK, p38, and JNK normalized to the total amount of the same proteins. (e) The cell viability of 3D culture A549 spheroids when treated with PAM, SB203580, SB202580 + PAM, PD98059, or PD98059 + PAM was shown as relative expression values. (f)The qRT-PCR results of EMT gene markers in 3D culture A549 spheroids when treated with PAM, SB203580, SB202580 + PAM, PD98059, or PD98059 + PAM were shown as relative expression values. The results represent average plus s.d. from at least three independent experiments. , , and ; ns: not significant. (g) Schematic representation of PAM in the regulation of cell survival and migration via PI3K/Akt and MAPKs pathways in 3D-cultured lung adenocarcinoma cells.

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