Effect of nucleolar P120 expression level on the proliferation capacity of breast cancer cells.
Steady-state level of nucleolar P120 protein and P120 mRNA was compared to the doubling time and S-phase fraction in human breast cancer cell lines growing exponentially and in similar cells treated with a single dose of P120 antisense oligodeoxynucleotides. The study included six breast cancer cell lines and one nontransformed breast cell line with doubling times from 1.1 to 5.5 days and with S-phase fractions from 35 to 9%. P120 expression level was determined by densitometric computerized evaluation of protein and mRNA blots and with a quantitative 32P-reverse transcriptase-polymerase chain reaction method developed for small-scale samples. In the slowest growing normal cell line, P120 expression level was only about 10% of the level found in the most rapidly growing cancer cell line. The amount of P120 mRNA was highly correlated with the amount of P120 protein (P = 0.0001), indicating that P120 accumulation is regulated in these cells primarily at a transcriptional level. There was also a significant positive correlation between the level of P120 protein/mRNA and doubling time of cell lines (P = 0.0008) or percentage of S-phase cells (P = 0.210). P120 antisense oligomer treatment decreased the growth rate of cells in a dose-dependent manner, and the inhibition reached 70% at 100 microM concentration. Both P120 mRNA and P120 protein levels were also decreased by approximately 70% in cells treated with 100 microM P120 antisense oligomer. Slowly growing cells exhibited 50% inhibition by treatment at a proportionally lower concentration of P120 antisense oligomer than fast growing cells. This study shows that the expression of P120, measured either at the protein or the mRNA level, correlates with proliferation rate, identifying P120 as a cell proliferation marker.
Fonagy
A
A
Department of Surgery, Lucille Parkey Markey Cancer Center, Lexington, Kentucky 40536.
Swiderski
C
C
Ostrovsky
A M
AM
Bolton
W E
WE
Freeman
J W
JW
D000951:Antigens, Neoplasm / Q000096:biosynthesis; D001483:Base Sequence; D001943:Breast Neoplasms; D002455:Cell Division; D002460:Cell Line; D005260:Female; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D014407:Tumor Cells, Cultured; D012359:tRNA Methyltransferases
D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C065028:NOP2 protein, human; D012359:tRNA Methyltransferases