GEN-27, a Newly Synthetic Isoflavonoid, Inhibits the Proliferation of Colon Cancer Cells in Inflammation Microenvironment by Suppressing NF-κB Pathway

Figure 4

Effects of GEN-27 on LPS-induced production of proinflammatory cytokines in THP-1 cells and the proliferation of HCT116 cells in response to the stimulation by THP-1-conditioned medium. (a) Human THP-1 cells were incubated with 10 μg/mL LPS for 24 h in the absence or presence of 1, 5, and 10 μM GEN-27 and 100 μM genistein. The expression of IL-6 and IL-1β mRNA in THP-1 cells was assessed by real-time PCR. GAPDH was used as an endogenous housekeeping gene. (b) The secretion levels of IL-6 and IL-1β in THP-1 cells after treatments of indicated doses of GEN-27 and genistein were assessed by ELISA. (c and d) HCT116 cells were either left untreated or treated with 10 μg/mL LPS, or THP-1-conditioned medium and indicated dose of GEN-27 together, or combination of 3 μg/mL IL-1RA, THP-1-conditioned medium, and indicated dose of GEN-27 for 24 h. Cell viability was assessed using an MTT assay and the results are expressed as the percentage of surviving cells over control cells. (e and f) The expressions of total proteins PCNA, bcl-2, and cyclin D1 were assessed by Western blot. The relative expressions of total proteins PCNA, bcl-2, and cyclin D1 were normalized to β-actin. Each value indicates the means ± SDs and is representative of the results obtained from three independent experiments. and compared with control; and versus LPS alone.

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