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. 2015 Jul 6;3(3):519–543. doi: 10.3390/vaccines3030519

Figure 1.

Figure 1

Effects of co-targeting HER-1 and HER-2 in EC or HER-1 and IGF-1R in TNBC. MTT proliferation assay shows significant inhibition of OE19 cells following single and combination treatment with HER-1-418 and HER-2-266 or HER-2-597 peptide vaccine antibodies (A) or peptide mimics (B) and significant inhibition of TNBC cells following single and combination treatment with HER-1-418 and IGF-1R peptide vaccine antibodies (C) or peptide mimcs (D). Cells were treated with peptide mimics or peptide vaccine antibodies for 1 h prior to ligand stimulation with EGF/HRG (50 ng/mL). After 72 h of incubation in the presence of the peptide mimics, MTT was used to measure cell proliferation. Percent inhibition was calculated by taking absorbance (abs) readings at 570 nm and using the following equation: (abs. untreated-abs. treated)/abs. untreated × 100). An irrelevant (IRR) peptide or normal rabbit IgG was used as a negative control; trastuzumab and cetuximab were positive controls. Values represent the mean of at least two independent experiments performed in triplicate (n = 3); error bars indicate SD of the mean. Single treatment with either peptide vaccine antibodies or peptide mimics in EC significantly inhibited proliferation over negative control (* p < 0.05). The anti-HER-2-597 and anti-HER-1-418 combination (A) also showed significantly higher inhibition compared to single treatment alone (* p < 0.05) and combination treatment with both HER-2-266 and HER-1-418 and HER-2-597 and HER-1-418 peptide mimics (B) inhibited proliferation more than single treatment alone (* p < 0.05). Combination treatment in TNBC cells with anti-HER-1-418 and anti-IGF-1R-56 (C) showed greater inhibition of cell proliferation over single treatment alone (# p < 0.005) as compared to the anti-HER-1-418 with anti-IGF-1R-233 and anti-IGF-1R-56 with anti-IGF-1R-233 combinations, which significantly inhibited cell proliferation over negative control (**p < 0.001) but did not show a significant advantage over single treatment alone. Similarly, the HER-1-418 and IGF-1R-56 peptide mimics (D) showed that the combination treatment inhibited proliferation of TNBC cells significantly more that single treatment alone (* p < 0.005).