Cell-cycle regulation of DNA damage-induced expression of the suppressor gene PML.
The promyelocytic leukemia (PML) gene, which encodes a growth- and transformation-suppressor, has been identified at the non-random chromosomal translocation break point t(15; 17)(q22; q12) of acute promyelocytic leukemia. To elucidate if PML may play a role in cellular response to DNA damage, PML expression was analyzed by immunofluorescence staining in HeLa cells treated with ionizing radiation (IR) and cisplatin. Our studies demonstrated IR at 20Gy, and cisplatin at 6 micrograms/ml caused more than 5-10 fold increases in PML protein expression in the PML Oncogenic Domain (POD) by immunofluorescent staining. Northern blotting showed that there was no gross increase in mRNA levels indicating that the induction is a post-transcriptional event. Flow cytometry showed that HeLa cells treated with IR were progressively arrested in G1, which correlates with the optimal expression of PML in the cell cycle. To determine if PML expression was under the control of the tumor suppressor p53, which is known to arrest cells in G1, HeLa cells were transfected with the wild-type p53 gene. PML expression in p53 transduced cells were 5-10 fold higher than the control, indicating that the enhanced expression of PML is apparently dependent on the p53 pathway. These data also indicate that PML may play an important role in cellular response to DNA damage such as DNA repair or apoptosis during G1 arrest.
Chan
J Y
JY
Department of Clinical Oncology, Chinese University of Hong Kong, Sir Y. K. Pao Centre for Cancer, Prince of Wales Hospital, Shatin. chanyhjohn@cujk.edu.hk
Li
L
L
Fan
Y H
YH
Mu
Z M
ZM
Zhang
W W
WW
Chang
K S
KS
D015152:Blotting, Northern; D015153:Blotting, Western; D002453:Cell Cycle; D002945:Cisplatin / Q000494:pharmacology; D004249:DNA Damage / Q000235:genetics*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D016193:G1 Phase / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D016158:Genes, p53 / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D011003:Ploidies; D000071156:Promyelocytic Leukemia Protein; D012333:RNA, Messenger / Q000032:analysis; D011839:Radiation, Ionizing; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000235:genetics; D025521:Tumor Suppressor Proteins
D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D012333:RNA, Messenger; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D002945:Cisplatin