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Basic Sciences

Autocrine Growth Factor in Defined Serum-free Medium of Human Salivary Gland Adenocarcinoma Cell Line HSG

Riki Kurokawa, Seiko Kyakumoto and Minoru Ota
Riki Kurokawa
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Seiko Kyakumoto
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Minoru Ota
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DOI:  Published September 1989
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Abstract

Human salivary gland adenocarcinoma cell line HSG secretes an epidermal growth factor (EGF)-like molecule and contains EGF receptors. Growth of HSG cells is inhibited by glucocorticoid. We have identified that the growth inhibition by glucocorticoid is induced by the reduced secretion of the EGF-like molecule and that addition of anti-human EGF antibody to the culture specifically inhibits the growth of HSG cells, suggesting that autocrine secretion is involved in the growth of HSG cells. To prove that autocrine secretion functions in glucocorticoid-regulated growth of the HSG cell line, we purified the EGF-like molecule from serum-free, defined medium conditioned by the HSG cells and examined the growth-stimulatory effect of the purified molecule. The cultivation of HSG cells in serum-free defined medium, which contains insulin (10 µg/ml) and transferrin (10 µg/ml) only as proteinaceous components, resulted in establishment of a new cell line (HSG-SF) which had different morphological features from the parental HSG cell line. HSG-SF cells were found to have basically the same responsiveness to glucocorticoid as parental HSG cells. Parental HSG cells secreted high molecular weight EGF-like molecules (Mr 46,000 and 57,000), which were recognized by specific antibody to low molecular weight human EGF (Mr 6,201). From conditioned, serum-free medium of HSG-SF cells, an EGF-like molecule (Mr 46,000) was purified by using an anti-human EGF antibody-coupled Sepharose CL-4B column. This EGF-like molecule induced a maximal increase (36%) in incorporation of [3H]thymidine into DNA of parental HSG cells as well as low molecular weight human EGF. These observations demonstrate that growth of the HSG cell line is regulated by autocrine secretion.

Footnotes

  • ↵1 This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan, Grants 63771501 (to R. K.), 63771500 (to S. K.), and 61480392 (to M. O.), and a grant from the Science Research Promotion Fund of the Japan Private School Promotion Foundation.

  • ↵2 To whom requests for reprints should be addressed.

  • Received March 31, 1989.
  • Revision received June 1, 1989.
  • Accepted June 16, 1989.
  • ©1989 American Association for Cancer Research.
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September 1989
Volume 49, Issue 18
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Autocrine Growth Factor in Defined Serum-free Medium of Human Salivary Gland Adenocarcinoma Cell Line HSG
Riki Kurokawa, Seiko Kyakumoto and Minoru Ota
Cancer Res September 15 1989 (49) (18) 5136-5142;

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Autocrine Growth Factor in Defined Serum-free Medium of Human Salivary Gland Adenocarcinoma Cell Line HSG
Riki Kurokawa, Seiko Kyakumoto and Minoru Ota
Cancer Res September 15 1989 (49) (18) 5136-5142;
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