Article Figures & Data
Figures
- Figure 1.
Ionizing radiation (IR) induces activation of the JAK2/STAT3/Bcl2/Bcl-XL survival signaling pathway in various human lung cancer cells. A549, H358, H157, and H292 cells were treated with increasing doses of IR. After 2 hours, levels of pJAK2 (Tyr1007/1008), pSTAT3 (Tyr705), Bcl2, Bcl-XL, and Mcl-1 were analyzed by Western blot analysis.
- Figure 2.
Persistent activation of the JAK2/STAT3/Bcl2/Bcl-XL pathway in human lung cancer cells is associated with acquired radioresistance. A, levels of pJAK2 (Tyr1007/1008), pSTAT3 (Tyr705), Bcl2, Bcl-XL, Mcl-1, and K-Ras were compared in various parental (A549, H358, and H157) and acquired radioresistant (A549-IRR, H358-IRR, and H157-IRR) human lung cancer cells. B, various parental and acquired radioresistant human lung cancer cells were treated with ionizing radiation (IR; 5 Gy). Cell growth was analyzed by colony formation assay as described in Materials and Methods. Reported values are the mean ± SD for three separate experiments.
- Figure 3.
STAT3 is accumulated in the nucleus of acquired radioresistant human lung cancer cells. STAT3 was analyzed by immunofluorescence staining using STAT3 antibody in A549, A549-IRR, H358, H358-IRR, H157, and H157-IRR cells.
- Figure 4.
Niclosamide (Niclo) blocks ionizing radiation (IR)-induced activation of the STAT3/Bcl2/Bcl-XL pathway and reverses acquired radioresistance of human lung cancer cells. A, A549, H358, and H157 cells were treated with IR (2 Gy) in the absence or presence of Niclo (1 μmol/L) for 24 hours. Levels of pJAK2 (Tyr1007/1008), pSTAT3 (Tyr705), Bcl2, Bcl-XL, Mcl-1, p-mTOR, p-P70S6K, p-4EBP1, and active caspase-3 were analyzed by Western blot analysis. B, radioresistant human lung cancer A549-IRR and H157-IRR cells were treated with increasing concentrations of Niclo for 24 hours. Levels of pJAK2 (Tyr1007/1008), pSTAT3 (Tyr705), Bcl2, Bcl-XL, Mcl-1, p-mTOR, p-P70S6K, p-4EBP1, and active caspase-3 were analyzed by Western blot analysis. C, A549 and A549-IRR cells were treated with IR (5 Gy), Niclo (0.1 μmol/L), or in combination. Cell growth was analyzed by colony formation assay. Results represent the mean ± SD for three separate determinations.
- Figure 5.
Specific knockdown of STAT3 reverses radioresistance of human lung cancer cells. A, STAT3 shRNA or control shRNA was transfected into A549 or A549-IRR cells. Expression levels of STAT3, STAT1, STAT5, Bcl-XL, and Bcl2 were analyzed by Western blot analysis. B and C, A549 or A549-IRR cells expressing STAT3 shRNA or control shRNA were treated with 5 Gy of ionizing radiation (IR). Cell growth was analyzed by colony formation assay. Results represent the mean ± SD for three separate determinations. D, A549-IRR cells were transfected with control shRNA or STAT3 shRNA. After 48 hours, cells were then treated with IR (2 Gy) twice, followed by colony formation assay.
- Figure 6.
Niclosamide overcomes acquired radioresistance in lung cancer xenografts. A, mice bearing lung cancer A549 or radioresistant A549-IRR xenografts were treated with vehicle control, ionizing radiation (IR; 2 Gy, twice per week), Niclo (30 mg/kg/d), or in combination for 21 days. Tumor volume was measured every other day. B and C, tumor tissues were removed at end of treatments. Active caspase-3 was analyzed by IHC staining using anti–active caspase-3 antibody (B). Protein expression levels of pSTAT3 (Tyr705), STAT3, Bcl2, Bcl-XL, Mcl-1, PARP, and active caspase-3 in tumor tissues were analyzed by Western blot analysis (C).
- Figure 7.
Toxicity analysis for treatments with ionizing radiation (IR) and niclosamide in mice bearing A549 xenografts. A, body weight of mice with A549 xenografts was measured once every other day during treatment with vehicle control, IR (2 Gy, twice per week), Niclo (30 mg/kg/d), or in combination. B, blood analysis of mice after various treatments for 21 days. C, H&E histology of various organs after various treatments for 21 days.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Figure 1 - PDF file - 218K, Ionizing radiation (IR) induces activation of the JAK2/STAT3/Bcl2/Bcl-XL survival signaling pathway in various human lung cancer cells.
- Supplementary Figure 2 - PDF file - 74K, Niclosamide (Niclo) reduces STAT3 nuclear localization in radioresistant human lung cancer A549-IRR cells.
- Supplementary Figure 3 - PDF file - 190K, Niclosamide increases the proportion of the cell population in the G0/G1 phase in A549 parental and radioresistant A549 (A549-IRR) cells.
- Supplementary Figure 4 - PDF file - 109K, Niclosamide-induced growth inhibition is diminished after specific knockdown of STAT3.
- Supplementary Figure 5 - PDF file - 142K, Niclosamide or in combination with IR inhibits cell proliferation in tumor tissues.
- Supplementary Figure 6 - PDF file - 181K, Toxicity analysis for treatments with IR and niclosamide in mice bearing A549-IRR xenografts.
Volume 13, Issue 3
