Relationship of p15 and p16 gene alterations to elevated dihydrofolate reductase in childhood acute lymphoblastic leukaemia.
The downstream effects of p15 and p16 gene deletions and loss of transcripts on dihydrofolate reductase (DHFR) were examined in 63 B-precursor (BP) acute lymphoblastic leukaemia (ALL) samples. p15 and/or p16 gene deletions were seen in 6% and 8%, respectively, of BP-ALL samples; however, losses of p15 and/or p16 transcripts were seen in 26 out of 63 (41%) samples. Loss of p15 transcripts (36.5%) exceeded that for p16 (17.5%). For the 26 BP-ALLs that lacked p15 and/or p16 transcripts, only six (23%) exhibited low levels of DHFR by flow cytometry assay with Pt430, a fluorescent anti-folate. Conversely, 18 out of 37 (49%) BP-ALL samples with intact p15 and/or p16 genes and transcripts showed low levels of DHFR (P = 0.04). In p15- and p16-null K562 cells transfected with a tetracycline-inducible p15 cDNA construct, induction of p15 transcripts and protein was accompanied by decreased growth rates, decreased S-phase fraction, decreased retinoblastoma protein phosphorylation, and markedly reduced levels of DHFR transcripts and protein. Collectively, our results suggest that losses of p15 and/or p16 gene expression result in elevated levels of DHFR in BP-ALL in children. However, additional downstream factors undoubtedly also contribute to elevated levels of this enzyme target.
Shah
S J
SJ
Experimental and Clinical Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Detroit, Michigan, USA.
Taub
J W
JW
Witt
T L
TL
Pollock
B H
BH
Ding
B C
BC
Moore
D S
DS
Amylon
M
M
Pullen
J
J
Ravindranath
Y
Y
Matherly
L H
LH
D000293:Adolescent; D015139:Blotting, Southern; D002051:Burkitt Lymphoma / Q000201:enzymology / Q000235:genetics*; D016022:Case-Control Studies; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002648:Child; D002675:Child, Preschool; D016001:Confidence Intervals; D050762:Cyclin-Dependent Kinase Inhibitor p15; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D017353:Gene Deletion; D015870:Gene Expression / Q000187:drug effects; D019942:Genes, p16; D006801:Humans; D007223:Infant; D020014:K562 Cells; D016015:Logistic Models; D008297:Male; D016017:Odds Ratio; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D025521:Tumor Suppressor Proteins
C493966:CDKN2B protein, human; D018797:Cell Cycle Proteins; D050762:Cyclin-Dependent Kinase Inhibitor p15; D012333:RNA, Messenger; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D013762:Tetrahydrofolate Dehydrogenase; D004318:Doxycycline