Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2003 Feb;9(2):271-5.
doi: 10.3748/wjg.v9.i2.271.

Study on the mechanism of epidermal growth factor-induced proliferation of hepatoma cells

Bin-Wen Wu  1 , Yuan Wu, Jia-Long Wang, Ju-Sheng Lin, Shu-Yu Yuan, Ai Li, Wu-Ren Cui
Affiliations

Study on the mechanism of epidermal growth factor-induced proliferation of hepatoma cells

Bin-Wen Wu et al. World J Gastroenterol. 2003 Feb.

Abstract

Aim: Many growth factors, such as epidermal growth factor (EGF), are associated with the carcinogenesis. EGF plays its role in the proliferation of hepatoma cells through binding with EGF receptor (EGFR) and a series of signal transduction. But the postreceptor pathway is still not clear. In the present experiment, we studied the effect of tyrosine kinase, protein kinase C, Na(+)/H(+) exchange, calmodulin and voltage-dependent Ca(2+) channel on EGF-induced hepatoma cell proliferation.

Methods: Hepatoma cell line SMMC7721 was cultured in RPMI1640 serum-free medium. In order to study the effect of thyrosine kinase, protein kinase C, Na(+)/H(+) exchange, calmodulin and voltage-dependent Ca(2+) channel on human heptoma cell proliferation induced by epidermal growth factor (EGF), DNA synthesis rate of hepatoma cells was measured by the method of (3)H-TdR incorporation.

Results: EGF (10(-9) M) stimulated the proliferation of heptoma cells significantly ((3)H-TdR incorporation was 1 880+/-281 cpm/well, P<0.05), and this effect was significantly inhibited by tyrosine kinase inhibitor genistein ((3)H-TdR incorporation was 808+/-209 cpm/well, P<0.001). Calmodulin inhibitor W-7, protein kinase C inhibitor H-7 and Na(+)/H(+) exchange inhibitor amiloride individually had significant inhibiting effect on EGF-induced proliferation of hepatoma cells ((3)H-TdR incorporation was 978+/-87.3 cpm/well, 1 241+/-147 cpm/well, 1 380+/-189 cpm/well, respectively, P<0.001, P<0.01, P<0.05), but they all had no effect on the basal level proliferation of cultured hepatoma cells ((3)H-TdR incorporation was 1 284+/-260 cpm/well, 1 179+/-150 cpm/well, 1 392+/-152 cpm/well, respectively, (3)H-TdR incorporation of the control was 1 353+/-175 cpm/well, P>0.05). Voltage-dependent Ca(2+) channel inhibitor verapamil had no inhibition on EGF-induced proliferation of hepatoma cells ((3)H-TdR incorporation was 1 637+/-133 cpm/well, P>0.05), it also had no effect on the basal level proliferation of cultured hepatoma cells ((3)H-TdR incorporation was 1 196+/-112 cpm/well, P>0.05).

Conclusion: Our data suggest that tyrosine kinase, Ca(2+)-calmodulin-dependent pathway, protein kinase C and Na(+)/H(+) exchange play a critical role in EGF-induced proliferation of hepatoma cells and that the effect of EGF is independent of voltage-dependent Ca(2+) channel.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Effect of genistein on EGF-induced 3H-TdR incorporation into hepatoma cells. n = 6, -x ± s, aP < 0.05, versus control; cP < 0.001, versus EGF 10-9M.
Figure 2
Figure 2
Effect of W-7 on EGF-induced 3H-TdR incorporation into hepatoma cells. n = 6, -x ± s, aP < 0.001 vs W-7 group; cP > 0.05 versus control.
Figure 3
Figure 3
Effect of verapamil on EGF-induced 3H-TdR incorporation into hepatoma cells. n = 6, -x ± s, aP > 0.05 vs EGF 10-9M group; cP > 0.05 versus control.
Figure 4
Figure 4
Effect of H-7 on EGF-induced 3H-TdR incorporation into hepatoma cells. n = 6, -x ± s, aP > 0.05 compared with control; bP < 0.01 vs EGF 10-9M group; cP < 0.05 versus control.
Figure 5
Figure 5
Effect of amiloride on EGF-induced 3H-TdR incorporation into hepatoma cells. n = 6, -x ± s, aP > 0.05 versus control; cP < 0.05 vs EGF (10-9M) group.
See this image and copyright information in PMC

References

    1. Diehl AM, Rai RM. Liver regeneration 3: Regulation of signal transduction during liver regeneration. FASEB J. 1996;10:215–227. - PubMed
    1. Fausto N, Laird AD, Webber EM. Liver regeneration. 2. Role of growth factors and cytokines in hepatic regeneration. FASEB J. 1995;9:1527–1536. - PubMed
    1. Michalopoulos GK, DeFrances MC. Liver regeneration. Science. 1997;276:60–66. - PubMed
    1. Yan JP, Jia JB, Ma XH, Wu XR, Zhao YC, Han DW. Immunohistochemical study on expression of epidermal growth factor receptor at hepatocyte nuclei in experimental rat liver cirrhosis. Huaren Xiaohua Zazhi. 1998;6:15–16.
    1. Kong YG, Zhen J, Lu GJ, Xu SB, Chen YF. The effects of EGF on the growth of human hepatoma cells and the regulatory action of somatostatin on EGF receptors. Zhonghua Xiaohua Zazhi. 1995;15:129–132.

MeSH terms