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Tyrosine-kinase expression profiles in human gastric cancer cell lines and their modulations with retinoic acids.

Kao HW, Chen HC, Wu CW, Lin WC - Br. J. Cancer (2003)

Bottom Line: Many protein tyrosine kinases are key regulators involved in cellular growth, differentiation, development, apoptosis and signal transduction pathways.Expression of tyrosine kinases in retionic acid-treated cancer cells was investigated by reverse trancriptase-polymerase chain reaction (RT-PCR) and a novel restriction analysis of gene expression (RAGE) display technique.In cells treated with 9-cis-retinoic acid or all-trans-retinoic acid, we found that two PTKs (Eph and Hek5) appeared to be upregulated.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC.

ABSTRACT
Many protein tyrosine kinases are key regulators involved in cellular growth, differentiation, development, apoptosis and signal transduction pathways. Obtaining a comprehensive tyrosine-kinase expression profile in tumour cells is essential to learning more about their oncogenic potentials and responses to various chemotherapeutic reagents - such as retinoic acid, which has been shown to suppress the growth of gastric cancer cells and modulate gene expression. Expression of tyrosine kinases in retionic acid-treated cancer cells was investigated by reverse trancriptase-polymerase chain reaction (RT-PCR) and a novel restriction analysis of gene expression (RAGE) display technique. We first established comprehensive tyrosine-kinase profiles in different human gastric cancer cell lines. In cells treated with 9-cis-retinoic acid or all-trans-retinoic acid, we found that two PTKs (Eph and Hek5) appeared to be upregulated. In the present study, we demonstrate an efficient and simple RAGE approach for examining tyrosine kinases' expression in tumour cells and their alterations following drug treatments.

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Related in: MedlinePlus

RAGE PTK profiling of different human tissues. Poly(A) mRNA obtained from Clontech was amplified by RT–PCR as described in Materials and Methods. The 150–170 bp amplicon was purified and used for MwoI digestion. The completed digested products were separated by a denaturing sequencing gel. Specific PTKs were identified by the digested fragment sizes as indicated.
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fig1: RAGE PTK profiling of different human tissues. Poly(A) mRNA obtained from Clontech was amplified by RT–PCR as described in Materials and Methods. The 150–170 bp amplicon was purified and used for MwoI digestion. The completed digested products were separated by a denaturing sequencing gel. Specific PTKs were identified by the digested fragment sizes as indicated.

Mentions: By using degenerated primers from submotifs VII and IX of the kinase domain as well as RT–PCR, we were able to easily amplify PTK genes expressed in cells. In addition to the previous ‘cloning and sequencing’ method, we utilised an improved RAGE method to provide a more efficient, economical and expeditious tyrosine-kinase profiling approach. We first established the PTK RAGE protocol in our laboratory by using a panel of human tissues. Samples from 12 tissues could be displayed on a single sequencing gel; thus we could effectively screen all known human PTKs in a short period of time. As shown in Figure 1Figure 1


Tyrosine-kinase expression profiles in human gastric cancer cell lines and their modulations with retinoic acids.

Kao HW, Chen HC, Wu CW, Lin WC - Br. J. Cancer (2003)

RAGE PTK profiling of different human tissues. Poly(A) mRNA obtained from Clontech was amplified by RT–PCR as described in Materials and Methods. The 150–170 bp amplicon was purified and used for MwoI digestion. The completed digested products were separated by a denaturing sequencing gel. Specific PTKs were identified by the digested fragment sizes as indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376380&req=5

fig1: RAGE PTK profiling of different human tissues. Poly(A) mRNA obtained from Clontech was amplified by RT–PCR as described in Materials and Methods. The 150–170 bp amplicon was purified and used for MwoI digestion. The completed digested products were separated by a denaturing sequencing gel. Specific PTKs were identified by the digested fragment sizes as indicated.
Mentions: By using degenerated primers from submotifs VII and IX of the kinase domain as well as RT–PCR, we were able to easily amplify PTK genes expressed in cells. In addition to the previous ‘cloning and sequencing’ method, we utilised an improved RAGE method to provide a more efficient, economical and expeditious tyrosine-kinase profiling approach. We first established the PTK RAGE protocol in our laboratory by using a panel of human tissues. Samples from 12 tissues could be displayed on a single sequencing gel; thus we could effectively screen all known human PTKs in a short period of time. As shown in Figure 1Figure 1

Bottom Line: Many protein tyrosine kinases are key regulators involved in cellular growth, differentiation, development, apoptosis and signal transduction pathways.Expression of tyrosine kinases in retionic acid-treated cancer cells was investigated by reverse trancriptase-polymerase chain reaction (RT-PCR) and a novel restriction analysis of gene expression (RAGE) display technique.In cells treated with 9-cis-retinoic acid or all-trans-retinoic acid, we found that two PTKs (Eph and Hek5) appeared to be upregulated.

View Article: PubMed Central - PubMed

Affiliation: Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan, ROC.

ABSTRACT
Many protein tyrosine kinases are key regulators involved in cellular growth, differentiation, development, apoptosis and signal transduction pathways. Obtaining a comprehensive tyrosine-kinase expression profile in tumour cells is essential to learning more about their oncogenic potentials and responses to various chemotherapeutic reagents - such as retinoic acid, which has been shown to suppress the growth of gastric cancer cells and modulate gene expression. Expression of tyrosine kinases in retionic acid-treated cancer cells was investigated by reverse trancriptase-polymerase chain reaction (RT-PCR) and a novel restriction analysis of gene expression (RAGE) display technique. We first established comprehensive tyrosine-kinase profiles in different human gastric cancer cell lines. In cells treated with 9-cis-retinoic acid or all-trans-retinoic acid, we found that two PTKs (Eph and Hek5) appeared to be upregulated. In the present study, we demonstrate an efficient and simple RAGE approach for examining tyrosine kinases' expression in tumour cells and their alterations following drug treatments.

Show MeSH
Related in: MedlinePlus