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. 2020 Sep 29;11(11):3117–3125. doi: 10.1111/1759-7714.13619

Figure 2.

Figure 2

Aspirin relieve the enhancement resistance, stemness and invasiveness of H460R cells in vitro. H460S and H460R with 0–5 μg/mL (0–17 μmol/L) or 0–50 μg/mL(0–170 μmol/L) of cisplatin, respectively, for 72 hours. Cell viability was evaluated by MTT assay. (a) H460R and H460S were assayed for cisplatin sensitivity (Inline graphic) H460S, (Inline graphic) H460R. (b) H460S was assayed for cisplatin sensitivity. Average and standard deviations (SD) of three independent experiments are presented. (c) Aspirin sensitivity of H460R after a 72 hours treatment. Cell viability was evaluated by MTT assay, 50% ± 3% survival at 16 μM. (d) H460S cells were stained with Annexin V‐APC and propidium iodide following treatment with or without 16 μM aspirin or 0.3 μg/mL cisplatin for 72 hours. Apoptosis was determined by flow cytometry. Cont, control. Regulation of proteins affecting (e) apoptosis, (f) drug resistance, (g) stemness; and (h) invasion and metastasis. Protein levels were detected by western blotting. β‐actin was used as the loading control. Numbers 1 and 4 were H460S cells treated without aspirin or cisplatin, and number 2, 3, 5, 6 were H460R cells. Number 2 was untreated. Numbers 3, 5, 6 were treated with aspirin, cisplatin, and aspirin with cisplatin, respectively. In the same strip, the density of 1 and 4 might not appear exactly the same because it was impossible for all parts of the strips to be applied to the same concentration of developer at the same time. We calculated the average value and SD of repeating the experiments for statistically significant times, so the density of 1 and 4 on the strip graph is slightly different, which does not affect our multiple experimental results. Values are expressed as means ± SD, n = 5, *P < 0.05.