Limits...
Receptor protein tyrosine kinase EphB4 is up-regulated in colon cancer.

Stephenson SA, Slomka S, Douglas EL, Hewett PJ, Hardingham JE - BMC Mol. Biol. (2001)

Bottom Line: We have used commercially available cDNA arrays to identify EphB4 as a gene that is up-regulated in colon cancer tissue when compared with matched normal tissue from the same patient.Quantitative RT-PCR analysis of the expression of the EphB4 gene has shown that its expression is increased in 82% of tumour samples when compared with the matched normal tissue from the same patient.The results presented here supports the emerging idea that Eph receptors play a role in tumour formation and suggests that further elucidation of this signalling pathway may identify useful targets for cancer treatment therapies.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Haematology, The Queen Elizabeth Hospital, Woodville Rd, Woodville, South Australia, Australia. sally.stephenson@eudoramail.com

ABSTRACT

Background: We have used commercially available cDNA arrays to identify EphB4 as a gene that is up-regulated in colon cancer tissue when compared with matched normal tissue from the same patient.

Results: Quantitative RT-PCR analysis of the expression of the EphB4 gene has shown that its expression is increased in 82% of tumour samples when compared with the matched normal tissue from the same patient. Using immunohistochemistry and Western analysis techniques with an EphB4-specific antibody, we also show that this receptor is expressed in the epithelial cells of the tumour tissue and either not at all, or in only low levels, in the normal tissue.

Conclusion: The results presented here supports the emerging idea that Eph receptors play a role in tumour formation and suggests that further elucidation of this signalling pathway may identify useful targets for cancer treatment therapies.

No MeSH data available.


Related in: MedlinePlus

(A) Coomassie stained duplicate gel for loading comparison. (B) Western analysis of EphB4 protein expression in colon tumour (T) and matched normal mucosa (N) from five different patients. The size markers in kDa are shown on the left. The arrow indicates the normal EphB4 protein.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC64642&req=5

Figure 4: (A) Coomassie stained duplicate gel for loading comparison. (B) Western analysis of EphB4 protein expression in colon tumour (T) and matched normal mucosa (N) from five different patients. The size markers in kDa are shown on the left. The arrow indicates the normal EphB4 protein.

Mentions: To determine if the increased expression of the EphB4 gene corresponded to an increase in the EphB4 protein we performed western analysis of colon and matched normal tissues using an EphB4-specific polyclonal antibody. The predicted size of wildtype EphB4 protein has been estimated by Bennet et al., (1994) to be 120 kD and in our experiment a band of this size was present in protein extracts from the tumour samples but low or absent in extracts from the normal mucosa (Figure 4) [28]. This confirms that the increased expression in the tumour tissues does correspond to an increase amount of the protein itself. Additional signals corresponding to proteins of lower molecular weight were also visible specifically in the tumour samples. It is unknown at this time whether these are EphB4-specific cleavage products or not and this will be the subject of future investigation. Further samples will be need to be tested to determine if there is any association between the stage of tumour development and amount of protein present.


Receptor protein tyrosine kinase EphB4 is up-regulated in colon cancer.

Stephenson SA, Slomka S, Douglas EL, Hewett PJ, Hardingham JE - BMC Mol. Biol. (2001)

(A) Coomassie stained duplicate gel for loading comparison. (B) Western analysis of EphB4 protein expression in colon tumour (T) and matched normal mucosa (N) from five different patients. The size markers in kDa are shown on the left. The arrow indicates the normal EphB4 protein.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC64642&req=5

Figure 4: (A) Coomassie stained duplicate gel for loading comparison. (B) Western analysis of EphB4 protein expression in colon tumour (T) and matched normal mucosa (N) from five different patients. The size markers in kDa are shown on the left. The arrow indicates the normal EphB4 protein.
Mentions: To determine if the increased expression of the EphB4 gene corresponded to an increase in the EphB4 protein we performed western analysis of colon and matched normal tissues using an EphB4-specific polyclonal antibody. The predicted size of wildtype EphB4 protein has been estimated by Bennet et al., (1994) to be 120 kD and in our experiment a band of this size was present in protein extracts from the tumour samples but low or absent in extracts from the normal mucosa (Figure 4) [28]. This confirms that the increased expression in the tumour tissues does correspond to an increase amount of the protein itself. Additional signals corresponding to proteins of lower molecular weight were also visible specifically in the tumour samples. It is unknown at this time whether these are EphB4-specific cleavage products or not and this will be the subject of future investigation. Further samples will be need to be tested to determine if there is any association between the stage of tumour development and amount of protein present.

Bottom Line: We have used commercially available cDNA arrays to identify EphB4 as a gene that is up-regulated in colon cancer tissue when compared with matched normal tissue from the same patient.Quantitative RT-PCR analysis of the expression of the EphB4 gene has shown that its expression is increased in 82% of tumour samples when compared with the matched normal tissue from the same patient.The results presented here supports the emerging idea that Eph receptors play a role in tumour formation and suggests that further elucidation of this signalling pathway may identify useful targets for cancer treatment therapies.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Haematology, The Queen Elizabeth Hospital, Woodville Rd, Woodville, South Australia, Australia. sally.stephenson@eudoramail.com

ABSTRACT

Background: We have used commercially available cDNA arrays to identify EphB4 as a gene that is up-regulated in colon cancer tissue when compared with matched normal tissue from the same patient.

Results: Quantitative RT-PCR analysis of the expression of the EphB4 gene has shown that its expression is increased in 82% of tumour samples when compared with the matched normal tissue from the same patient. Using immunohistochemistry and Western analysis techniques with an EphB4-specific antibody, we also show that this receptor is expressed in the epithelial cells of the tumour tissue and either not at all, or in only low levels, in the normal tissue.

Conclusion: The results presented here supports the emerging idea that Eph receptors play a role in tumour formation and suggests that further elucidation of this signalling pathway may identify useful targets for cancer treatment therapies.

No MeSH data available.


Related in: MedlinePlus