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. 1999 Jun;44(6):834–838. doi: 10.1136/gut.44.6.834

Expression and activity of mitogen activated protein kinases in human colorectal carcinoma

S Eggstein 1, M Franke 1, I Kutschka 1, G Manthey 1, B U von Specht 1, G Ruf 1, E Farthmann 1
PMCID: PMC1727542  PMID: 10323886

Abstract

BACKGROUND—Mitogen activated protein kinases (MAPKs) play a central role in the regulation of both cell growth and differentiation. They are involved in signal transduction of oncogenes and growth factors. The role of MAPK in colonic carcinoma is unknown.
AIMS—To establish whether the expression and activity of p42/44 MAPKs are altered in colorectal tumours as compared with normal mucosa.
METHODS—The expression and activity of p42/p44 MAPK were investigated in 22 colorectal carcinomas, four adenomas, and the corresponding normal colorectal mucosa by the use of western blotting, immunoprecipitation, and in vitro kinase assays.
RESULTS—After immunoprecipitation with an antibody specific for p42 MAPK, we found significant inactivation of p42 MAPK in colonic carcinomas as well as in adenomas, whereas most sample pairs showed only minor differences in p42 MAPK expression. Investigation of MAPK with an antibody capable of detecting both p42 and p44 MAPK showed a slight but significant decrease in p44 MAPK content in malignant tissues. With this antibody, only minor alterations in MAPK activity and no correlation with p42 MAPK activity were found.
CONCLUSIONS—Inactivation of p42 MAPK could be associated with colonic carcinogenesis.


Keywords: mitogen activated protein kinase (MAPK); Raf-1; colorectal cancer

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Figure 1  .

Figure 1  

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Activity of p42 mitogen activated protein kinase (MAPK) in colorectal cancer. The activities (seastar units (SSU)/mg protein) of corresponding colonic cancer/mucosa samples (n = 22) are connected by lines. Asterisks with error bars indicate the mean values and standard errors. Activities were examined by in vitro kinase assay after immunoprecipitation with sc-154.

Figure 2  .

Figure 2  

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Anti-mitogen activated protein kinase (MAPK) western blots of five representative paired samples of colon cancer (T) and mucosa (M). For the immunoblots shown in the upper row the sc-94 antibody, which detects p42 MAPK and p44 MAPK, was used. The blots shown below were obtained with p42 MAPK-specific sc-154.

Figure 3  .

Figure 3  

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Relative activity and expression of p42 mitogen activated protein kinase (MAPK) and p44/42 MAPK. The cancer/mucosa ratio of every sample pair is indicated. p42 MAPK was assayed by the use of sc-154 and is indicated by circles. p44/42 MAPK was determined using sc-94 and is displayed by diamonds.

Figure 4  .

Figure 4  

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Activity of p42 mitogen activated protein kinase (MAPK) in colonic adenomas. p42 MAPK activity was assayed in four colonic adenomas, two of them presenting together with carcinomas. (A) Autoradiographs of phosphorylated myelin basic protein; (B) bar graph of the activity (seastar units (SSU)/mg protein) quantified with a phosphoimager.

Figure 5  .

Figure 5  

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Relative expression of p44 mitogen activated protein kinase (MAPK) in colonic cancer. Immunoblots using sc-94 were quantified densitometrically. Points indicate the proportion of p44 MAPK ((p44 × 100)/(p44+p42)). The values (percentage) of corresponding colonic cancer/mucosa samples (n = 22) are connected by lines. Asterisks with error bars indicate the mean values and standard errors.

Figure 6  .

Figure 6  

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Raf-1 mobility shifts of three paired samples of colon cancer (T) and mucosa (M).

Selected References

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