Altered expression of nuclear matrix proteins in etoposide induced apoptosis in HL-60 cells.
The events of cell death and the expression of nuclear matrix protein (NMP) have been investigated in a promyelocytic leukemic cell line HL-60 induced with etoposide. By means of TUNEL assay, the nuclei displayed a characteristic morphology change, and the amount of apoptotic cells increased early and reached maximun about 39% after treatment with etoposide for 2 h. Nucleosomal DNA fragmentation was observed after treatment for 4 h. The morphological change of HL-60 cells, thus, occurred earlier than the appearance of DNA ladder. Total nuclear matrix proteins were analyzed by 2-dimensional gel electrophoresis. Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells. Western blotting was then performed on three nuclear matrix acssociated proteins, PML, HSC70 and NuMA. The expression of the suppressor PML protein and heat shock protein HSC70 were significantly upregulated after etoposide treatment, while NuMA, a nuclear mitotic apparatus protein, was down regulated. These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process.
Jin
M L
ML
College of Animal Science and Veterinary Medicine, Huazhong Agricultural University, Wuhan, China. jml8328@public.wh.hb.cn
Zhang
P
P
Ding
M X
MX
Yun
J P
JP
Chen
P F
PF
Chen
Y H
YH
Chew
Y Q
YQ
D034961:Antigens, Nuclear; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D018797:Cell Cycle Proteins; D004247:DNA / Q000032:analysis; D053938:DNA Fragmentation / Q000187:drug effects / Q000502:physiology; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005047:Etoposide / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000502:physiology*; D018922:HL-60 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D009363:Neoplasm Proteins / Q000187:drug effects / Q000378:metabolism; D015530:Nuclear Matrix / Q000187:drug effects / Q000378:metabolism*; D034521:Nuclear Matrix-Associated Proteins; D009687:Nuclear Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors / Q000187:drug effects / Q000378:metabolism; D025521:Tumor Suppressor Proteins
D034961:Antigens, Nuclear; D000972:Antineoplastic Agents, Phytogenic; D018797:Cell Cycle Proteins; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; C494371:HSPA8 protein, human; C050045:NUMA1 protein, human; D009363:Neoplasm Proteins; D034521:Nuclear Matrix-Associated Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; D005047:Etoposide; D004247:DNA