Electron microscopic identification of the intracellular secretion pathway of human G-CSF in a human tumor cell line: a comparative study with a Chinese hamster ovary cell line (IA1-7) transfected with human G-CSF cDNA.
Using monoclonal antibodies specific for human granulocyte colony-stimulating factor (G-CSF), intracellular localization of G-CSF in a G-CSF-producing human tumor cell line (CHU-2) and its ultrastructural characters were described and compared with those of a Chinese hamster ovary cell line (IA1-7) transfected with human G-CSF cDNA. The CHU-2 line, which was derived from a poorly differentiated squamous cell carcinoma of the oral cavity, preserved the character of a poorly differentiated squamous cell carcinoma. In the CHU-2 cell line, there were few cells immunohistochemically positive for G-CSF under light microscopic analysis despite the high transcription level of G-CSF cDNA and secretion of G-CSF that were comparable with cDNA-transfected IA1-7 cells. Using electron microscopy, the reaction products were localized mainly in the perinuclear space (PNS) and rough endoplasmic reticula (RER) without dilation of the cisternae, but they were very rarely found in the Golgi complex and not at all in other intracellular organelles. In contrast, most cells were positive for G-CSF in the IA1-7 cell line. Reaction products in this cell line were also demonstrated in the PNS and RER without dilation of the cisternae. These immunohistochemical findings, in conjunction with the results of Western and Northern blot analysis, suggested that G-CSF was secreted via the PNS and RER without intracellular retention.
Akatsuka
A
A
Division of Electron Microscopy, Tokai University School of Medicine, Kanagawa, Japan.
Shimamura
K
K
Katoh
Y
Y
Takekoshi
S
S
Nakamura
M
M
Nomura
H
H
Hasegawa
M
M
Ueyama
Y
Y
Tamaoki
N
N
D000818:Animals; D000911:Antibodies, Monoclonal; D015152:Blotting, Northern; D015153:Blotting, Western; D002451:Cell Compartmentation; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism; D015870:Gene Expression; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008854:Microscopy, Electron; D009368:Neoplasm Transplantation; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D016179:Granulocyte Colony-Stimulating Factor