Lung Krüppel-like factor (LKLF) is a transcriptional activator of the cytosolic phospholipase A2 alpha promoter.
Increased expression of cPLA2 (cytosolic phospholipase A2) has been shown to be the cause of tumorigenesis of NSCLC (non-small-cell lung cancer). Our laboratory has previously demonstrated that oncogenic forms of Ras increase transcription of cPLA2 in normal lung epithelial cells and NSCLC lines through activation of the ERK (extracellular-signal-regulated kinase) and JNK (c-Jun N-terminal kinase) MAPK (mitogen-activated protein kinase) family. We have also defined a minimal region of the cPLA2 promoter that is critical for this induction. To identify potential transcription factors that bind to this region and regulate expression, a yeast one-hybrid screen was performed with a rat lung cDNA library. Multiple members of the Krüppel family were identified, with LKLF (lung Krüppel-like factor) being isolated a number of times. Overexpression of LKLF in lung epithelial cells or Drosophila SL-2 cells increased cPLA2 promoter activity. Conversely, expression of a dominant negative form of LKLF inhibited induction of cPLA2 promoter activity by oncogenic Ras in normal lung epithelial cells and NSCLC. By electrophoretic mobility-shift assay analysis, it was found that LKLF bound to a GC-rich region of the cPLA2 promoter located between -37 and -30 upstream from the transcription start site. Expression of siRNA (small interfering RNA) directed against LKLF inhibited basal expression of cPLA2 in lung epithelial cells and blocked induction by H-Ras. In NSCLC, siRNA against LKLF co-operated with siRNA against Sp1 (stimulatory protein 1) to inhibit cPLA2 promoter activity. Finally, recombinant LKLF was a substrate for ERKs. These results indicate that LKLF is an important regulator of cPLA2 expression and participates in the induction of this protein, which is critical for increased eicosanoid production associated with lung tumorigenesis.
Wick
Marilee J
MJ
Department of Medicine, University of Colorado Health Sciences Center, 4200 E. 9th Ave., Denver, CO 80262, USA.
Blaine
Stacy
S
Van Putten
Vicki
V
Saavedra
Milene
M
Nemenoff
Raphael A
RA
D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D003600:Cytosol / Q000201:enzymology*; D004330:Drosophila / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004790:Enzyme Induction / Q000502:physiology; D004847:Epithelial Cells / Q000737:chemistry / Q000378:metabolism; D054513:Group IV Phospholipases A2; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D008168:Lung / Q000166:cytology; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D015534:Trans-Activators / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D014162:Transfection / Q000379:methods; D020798:Two-Hybrid System Techniques
C096161:KLF2 protein, human; D051741:Kruppel-Like Transcription Factors; D034741:RNA, Small Interfering; D015534:Trans-Activators; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2