Oyaksungisan, a Traditional Herbal Formula, Inhibits Cell Proliferation by Induction of Autophagy via JNK Activation in Human Colon Cancer Cells
Figure 3
Induction of autophagy by OY in HCT116 cells. (a) Cells were treated with indicated doses of OY for 24 h or treated with 500 μg/mL OY for indicated times before lysis. Western blot analyses were performed for checking the protein levels of LC3-I and -II. The band intensities were quantified by Image J program and are represented in the bar graph. β-Actin was used as a loading control for normalization. (b) To confirm the autophagy induction by OY, cells were treated with 1000 μg/mL OY for 24 h with or without pretreatment with 10 mM 3-MA for 1 h. After 24 h of OY exposure and 3-MA, cell morphology was observed under a phase contrast microscope. (c) Cells were treated with 500 μg/mL OY with or without preexisting 10 mM 3-MA for 3 h, and the upregulation of LC3-II level by OY was determined as a Western blot analysis. The band intensities were quantified by Image J program, and values indicate relative levels determined after normalization to untreated cells. (d) To confirm the autophagic effect of OY against cell proliferation on HCT116 cells, cells were treated with 1000 μg/mL OY with or without pre-existing 10 mM 3-MA for 24 h. After 24 h of OY exposure and 3-MA, cell viability was determined using MTT assay. Values indicate the mean ± SD. ** versus untreated control cells. * versus treated cells with OY only.
