Figure 3.

Effects of LPS pretreatment on cisplatin-induced apoptosis and NF-κB activation in cancer cells. PCI cell lines were treated with 1 μg/mL LPS. A, LPS pretreatment protects tumor cells from apoptosis induced by cisplatin. Cell lines were first incubated ± LPS for 12 h and then with 10 μmol/L cisplatin for 24 h, stained with 7-AAD, and examined by flow cytometry. Results of a representative experiment of three performed are shown. B, morphologic changes in culture of PCI-13 cells incubated ± cisplatin or + LPS and cisplatin. Magnification, ×200. C, Western blots of PCI-30 cells incubated ± LPS show activation of the PI3K/Akt survival pathway. Increased levels of MyD88 and IRAK-4 in cells stimulated with LPS are accompanied by nuclear localization of the p65 subunit of NF-κB in LPS-treated PCI-30 cells. D, tumor cells plated overnight were stimulated with LPS (0.1 μg/mL) for 12 h and then stained as described in Materials and Methods. 1 to 3, control cells were untreated; 1, 3, 4, and 6, nuclei are stained blue; 2 and 5, the p65 subunit of NF-κB is stained green; 3, an overlay of 1 and 2; 6, an overlay of 4 and 5. At least 200 cells were randomly counted. Bar, 20 μm. Results are representative of three independent experiments.