Fig. 8

Consequences of overexpression of SPDEF on the differentiation of immortalized hSABCi-NS1.1 cells on ALI, as a model of secretory cell hyperplasia in COPD. Based on the knowledge that SPDEF, a transcription factor that induced secretory cell differentiation is up-regulated in the SAE of COPD smokers. hSABCi-NS1.1 cells were genetically modified by lentivirus expressing SPDEF and cultured on ALI to day 14 to test whether the hSABCi-NS1.1 cells can be used as a model of secretory cell hyperplasia. a Secretory cell gene expression in the SAE of 36 COPD smokers compared to 60 healthy nonsmokers in vivo. Five up-regulated and 4 down-regulated secretory cell genes were used as examples. Data shown is relative fold-change of microarray-based gene expression ± standard error. b Staining of a known SPDEF-regulated gene in ALI. MUC5AC was used as an example. Shown are passage 47 cells on ALI-day 21. Red-MUC5AC; blue-nucleus. Top panel, lenti-GFP infected cells, as control; bottom panel, lenti-SPDEF-infected cells. Bar = 50 μm. c TaqMan PCR assessment of lentivirus-SPDEF-induced gene expression changes in hSABCi-NS1.1 cells. The same genes as in panel A were assessed. Data shown is the mean fold-change ± standard error of n = 3 independent experiments using passage 52 to passage 55 cells. d Staining of MSMB as an example of a SPDEF-regulated secretory gene in ALI. Shown are passage 47 cells on ALI-day 21. Red-MSMB; blue-nucleus. Top panel, lenti-GFP infected cells, as control; bottom panel, lenti-SPDEF-infected cells. For a, c *, p < 0.05; **, p < 0.01; ***, p < 0.001; #, consistent changes in all 3 independent experiments, but not significant because of variability