Fig. 8
From: Nestin regulates cellular redox homeostasis in lung cancer through the Keap1–Nrf2 feedback loop
Nestin/Nrf2 overexpression in NSCLC correlates with 17-AAG resistance. a IHC was performed to examine the relationship between Nestin expression and Nrf2 expression in 200 primary NSCLC specimens. The results obtained from two representative cases are shown (left). Percentages of specimens showing low or high Nestin expression relative to the levels of Nrf2 staining (right). Scale bar: 500 μm. b Nestin expression positively correlated with the expression levels of Nrf2-target genes (HO-1, NQO1, GCLM, and GCLC) in NSCLC specimens. The mRNA levels of Nestin and Nrf2-target genes were detected by qRT-PCR, and the correlations between the mRNA levels of Nestin and those of various antioxidant enzymes were evaluated using Pearson’s correlation coefficients (n = 12). c The expression levels of Nestin and Nrf2 were assessed in NSCLC tumors of grades I–IV. Scale bar: 100 μm. d The percentage of tumors showing simultaneous upregulation of Nestin and Nrf2 gradually increased from grade I to grade IV. e A549 cells were transfected with siNestin RNA or ovNestin plasmids. After 5 days, the cells (5 × 106 cells) were subcutaneously injected into nude mice (n = 4 per group), which were thereafter treated with 17-AAG three times a week. Tumor growth was examined by bioluminescent imaging on day 21. f Bioluminescent density was quantified. g Tumor weight on day 21 after mice were xenografted with NSCLC cells transfected with Control, shNestin, or ovNestin plasmids, with or without 17-AAG treatment. h Proposed model of the relationship between Nestin and Nrf2 in NSCLC cells. Data are presented as the means ± SD of at least three independent experiments. *P < 0.05, **P < 0.01, and ***P < 0.001, Student’s t test. Source data are available as a Source Data file
