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J BUON. 2019 Jan-Feb;24(1):116-122.

Effect of AMPK/Akt/mTOR pathway on cytokine-induced killer cells immunotherapy on colorectal cancer cells.

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1
Department of General Surgery, Tongde Hospital of Zhejiang Province, Hangzhou, China.

Abstract

PURPOSE:

To investigate the effect of cytokine-induced killer cells (CIKs) immunotherapy on colorectal cancer cells and the possible mechanisms using co-culture model of CIKs and human colorectal cancer cells.

METHODS:

CIKs were induced by INF-γ, CD3 and recombinant human IL-2 culture system in vitro. A co-culture model of CIKs and SW480 colorectal cancer cell line was established. The proliferation, invasion and apoptosis of SW480 cells were detected by CCK-8 (cell counting kit-8), transwell assay and Hoechst staining, respectively. The contents of IL-2, IL-10, insulin-like growth factor (IGF)-β and vascular endothelial growth factor (VEGF) in the culture medium was detected by ELISA (enzyme-linked immunosorbent assay). The expression levels of AMPK/Akt/mTOR signaling-related proteins were detected by Western blot.

RESULTS:

CCK-8 assay showed that the proliferative ability of co-cultured CIK+SW480 cells was remarkably lower than that of SW480 cells (p<0.05). Transwell assay showed that the number of invasive cells in co-cultured CIK+SW480 cells was less than that of SW480 cells (p<0.05). Hoechst staining showed that the apoptosis rate of co-cultured CIK+SW480 cells was higher compared with that of SW480 cells (p<0.05). Increased levels of IL-2 and IL-10 and decreased levels of IGF-β and VEGF were found in the co-culture medium by ELISA (p<0.05). Western blot results indicated that p-AMPK and p-Akt were upregulated, whereas FoxM1 and p-mTOR were downregulated in co-cultured CIK+SW480 cells compared with SW480 cells (p<0.05).

CONCLUSIONS:

CIKs inhibit the proliferation and invasion of colorectal cancer cells through downregulating FoxM1 and mTOR via AMPK/Akt/mTOR pathway.

PMID:
30941959

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