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Effects of EGF and TGF?1 onc-myc gene expression and DNA synthesis in embryonic hamster palate mesenchymal cells

✍ Scribed by Izadnegahdar, Mehrnaz F. ;Rathanaswami, Palaniswami ;Shah, Ravindra M.


Publisher
John Wiley and Sons
Year
1999
Tongue
English
Weight
237 KB
Volume
254
Category
Article
ISSN
0003-276X

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✦ Synopsis


Previous work has shown that cell proliferation is a major contributor to the early palate morphogenesis in mammals. The present study was undertaken to examine the effect of EGF, TGF␤ 1 and their combination on proliferation (measured by DNA synthesis) and on the expression of a growth related proto-oncogene, c-myc, in embryonic hamster palate mesenchymal cells (HPMC).

Vertically developing hamster palatal shelves were dissected on day 11 of gestation, and trypsinized, and primary cultures were grown in DMEM ϩ 10% serum at 37°C and 5% CO 2 . Following appropriate growth factor treatment of HPMC, DNA synthesis was measured by scintillation counting and extracted RNA was subjected to Northern blot analysis.

In serum-starved, pre-confuent cultures treated with EGF (20 ng/ml), DNA synthesis was stimulated in the presence of 2.5% serum. In contrast, treatment of HPMC with TGF␤ 1 (10 ng/ml) in the presence or absence of EGF/serum for 24 hr, or HPMC pre-treatment with TGF␤ 1 (30 min) followed by EGF/serum (24 hr), resulted in an arrest of DNA synthesis. Northern blot analysis of RNA extracted from HPMC showed that as serum-starved, growth-arrested cells progressed through G0 to G1 phase of the cell cycle, following EGF treatment, c-myc was expressed by 1 hr and declined thereafter. In contrast, TGF␤ 1 did not support expression of c-myc. Following pre-or co-treatment with TGF␤ 1 , the EGF Ϯ serum-induced expression of c-myc was seen between 1 and 6 hr.

It appears that EGF-induced expression of c-myc may be involved in advancing the HPMC in G1, and thus may contribute to the onset of DNA synthesis in HPMC. Since co-or pre-treatment with TGF␤ 1 did not inhibit EGF/serum induced expression of c-myc, it is possible that growth arresting effect of TGF␤ 1 may not be exerted directly through inhibition or blockage of c-myc expression.