Skip to main content
. 2021 Feb 5;22(4):1626. doi: 10.3390/ijms22041626

Figure 1.

Figure 1

Treatment of human lung cancer cell lines (A549, H292, H1299, and H358) with Hiltonol+anti-IL6+stattic+AG490 (Hiltonol+++) significantly suppressed cancer cell survival. (A) Dose optimization of Hiltonol and polyI:C (polyinosinic-polycytidilic acid) was performed and cell survival was assayed using MTT {3-(4,5-dimethylathiazol-2-yl)-2,5-diphenyl tetrazolium bromide} and CTB (CellTiter Blue). Both assays consistently showed that Hiltonol significantly suppressed the cell viability. The optimal killing dose of Hiltonol is 20 μg/mL (red line), compared to polyI:C (blue line) (**, p < 0.01). Differences between averages were analyzed by two-way ANOVA (two-way analysis of variance) with interaction. Doses and cell lines were considered as two factors in the two-way ANOVA. In MTT assays, LSD (Least Significant Difference) value in A549, H292, H1299, and H358 are 3.902, 3.334, 2.172, and 2.046, respectively. In CTB assays, LSD value in A549, H292, H1299, and H358 are 3.063, 2.939, 2.28, and 1.739, respectively. (B) Combinatorial treatments with Hiltonol+++ (red line) or with polyI:C+++ (blue line) further reduced cancer cell survival, with up to 1.7-fold greater killing by Hiltonol+++ compared to polyI:C+++ (***, p < 0.001). Data are representative of means ± SD (n = 3). For each time point, cell counts were normalized to corresponding DMSO (or PBS (phosphate buffered saline))-treated controls. In MTT assays, the LSD value in A549, H292, H1299, and H358 are 3.745, 2.341, 3.004, and 2.489, respectively. In CTB assays, the LSD value in A549, H292, H1299, and H358 are 3.222, 2.969, 3.423, and 3.396, respectively.