Secretion of polypeptides related to epidermal growth factor and insulinlike growth factor I by a human teratocarcinoma cell line |
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Authors: | Naihe Jing Robert Shiurba Hiroshi Kitani Hisaaki Kawakatsu Yasuhiro Tomooka Teruyo Sakakura |
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Institution: | (1) Laboratory of Cell Biology, Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), 3-1-1 Koyadai, 305 Tsukuba, Ibaraki, Japan;(2) Shanghai Institute of Biochemistry, 320 Yue-yang Road, 200031 Shanghai, China;(3) Present address: National Institute of Animal Health, 3-1-1 Kannondai, 305 Tsukuba, Ibaraki, Japan;(4) Present address: Nippon Shinyaku Co., Ltd., Nishiohji Hachijo, Minamiku, 601 Kyoto, Japan |
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Abstract: | Summary To identify polypeptide growth factors for human teratocarcinoma cells, we studied the malignant ovarian teratoma-derived
cell line, PA-1, that grew autonomously in serum-free medium. Medium conditioned by undifferentiated PA-1 cells strongly stimulated
proliferation of the mouse mammary tumor cell line, GR 2H6, which is responsive to epidermal growth factor (EGF) and insulinlike
growth factor-I (IGF-I). After ammonium sulfate precipitation, PA-1 conditioned medium was analyzed by anion exchange chromatography
and bioassay of elution fractions on GR 2H6 cells that were grown in medium deficient in either EGF or insulin. The results
demonstrated that PA-1 CM contained factors that can substitute for EGF and IGF-I in stimulating growth of GR 2H6 cells. Western
blots of peak mitogenic fractions revealed low molecular weight polypeptides that were immunoreactive with either anti-EGF
or anti-IGF-I antibodies. Indirect immunofluorescence staining of PA-1 cells with monoclonal antibodies localized receptors
for each growth factor, and binding of human EGF and IGF-I to these cells was quantified by radioreceptor assays. Secretion
of factors closely related to EGF and IGF-I by PA-1 cells under serum-free conditions may provide a novel model system to
study molecular mechanisms of autocrine growth stimulation in teratocarcinomas. |
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Keywords: | human teratocarcinoma EGF IGF-I |
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