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Comparative proteomics analysis of oral cancer cell lines: identification of cancer associated proteins.

Karsani SA, Saihen NA, Zain RB, Cheong SC, Abdul Rahman M - Proteome Sci (2014)

Bottom Line: Twenty four protein spots were found to have changed in abundance.Identified proteins were classified into seven functional categories - structural proteins, enzymes, regulatory proteins, chaperones and others.The mRNA expressions of two proteins - 14-3-3 protein sigma and Stress-induced-phosphoprotein 1 - were found to correlate with the corresponding proteins' abundance.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. saiful72@um.edu.my.

ABSTRACT

Background: A limiting factor in performing proteomics analysis on cancerous cells is the difficulty in obtaining sufficient amounts of starting material. Cell lines can be used as a simplified model system for studying changes that accompany tumorigenesis. This study used two-dimensional gel electrophoresis (2DE) to compare the whole cell proteome of oral cancer cell lines vs normal cells in an attempt to identify cancer associated proteins.

Results: Three primary cell cultures of normal cells with a limited lifespan without hTERT immortalization have been successfully established. 2DE was used to compare the whole cell proteome of these cells with that of three oral cancer cell lines. Twenty four protein spots were found to have changed in abundance. MALDI TOF/TOF was then used to determine the identity of these proteins. Identified proteins were classified into seven functional categories - structural proteins, enzymes, regulatory proteins, chaperones and others. IPA core analysis predicted that 18 proteins were related to cancer with involvements in hyperplasia, metastasis, invasion, growth and tumorigenesis. The mRNA expressions of two proteins - 14-3-3 protein sigma and Stress-induced-phosphoprotein 1 - were found to correlate with the corresponding proteins' abundance.

Conclusions: The outcome of this analysis demonstrated that a comparative study of whole cell proteome of cancer versus normal cell lines can be used to identify cancer associated proteins.

No MeSH data available.


Related in: MedlinePlus

Results of quantitative RT-PCR showing relative expressions of STMN1, 1433S, STIP1 and GSTP1. Total mRNA was extracted from normal and oral cancer cell lines. Quantitative RT-PCR was then performed as described. A: Relative expression of STMN1, B: Relative expression of 1433S, C: Relative expression of STIP1, D: Relative expression of GSTP1. Relative intensities of all genes of interest were determined by using β-actin as an internal standard. Results represent the mean ± SD for three experiments. *p < 0.05.
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Figure 3: Results of quantitative RT-PCR showing relative expressions of STMN1, 1433S, STIP1 and GSTP1. Total mRNA was extracted from normal and oral cancer cell lines. Quantitative RT-PCR was then performed as described. A: Relative expression of STMN1, B: Relative expression of 1433S, C: Relative expression of STIP1, D: Relative expression of GSTP1. Relative intensities of all genes of interest were determined by using β-actin as an internal standard. Results represent the mean ± SD for three experiments. *p < 0.05.

Mentions: Quantitative real-time PCR was performed to assess the mRNA expression of four proteins - STMN1, 1433S, STIP1 and GSTP1. These proteins were selected due the marked difference in their abundance and/or possible involvement in cancer. They also represented proteins from various functional categories. The mRNA expression of all four genes was found to correlate with its corresponding protein abundance. However, only 1433S and STIP1 was found to be statistically significant (Figure 3). This suggested that these proteins may be regulated at the mRNA level.


Comparative proteomics analysis of oral cancer cell lines: identification of cancer associated proteins.

Karsani SA, Saihen NA, Zain RB, Cheong SC, Abdul Rahman M - Proteome Sci (2014)

Results of quantitative RT-PCR showing relative expressions of STMN1, 1433S, STIP1 and GSTP1. Total mRNA was extracted from normal and oral cancer cell lines. Quantitative RT-PCR was then performed as described. A: Relative expression of STMN1, B: Relative expression of 1433S, C: Relative expression of STIP1, D: Relative expression of GSTP1. Relative intensities of all genes of interest were determined by using β-actin as an internal standard. Results represent the mean ± SD for three experiments. *p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3974152&req=5

Figure 3: Results of quantitative RT-PCR showing relative expressions of STMN1, 1433S, STIP1 and GSTP1. Total mRNA was extracted from normal and oral cancer cell lines. Quantitative RT-PCR was then performed as described. A: Relative expression of STMN1, B: Relative expression of 1433S, C: Relative expression of STIP1, D: Relative expression of GSTP1. Relative intensities of all genes of interest were determined by using β-actin as an internal standard. Results represent the mean ± SD for three experiments. *p < 0.05.
Mentions: Quantitative real-time PCR was performed to assess the mRNA expression of four proteins - STMN1, 1433S, STIP1 and GSTP1. These proteins were selected due the marked difference in their abundance and/or possible involvement in cancer. They also represented proteins from various functional categories. The mRNA expression of all four genes was found to correlate with its corresponding protein abundance. However, only 1433S and STIP1 was found to be statistically significant (Figure 3). This suggested that these proteins may be regulated at the mRNA level.

Bottom Line: Twenty four protein spots were found to have changed in abundance.Identified proteins were classified into seven functional categories - structural proteins, enzymes, regulatory proteins, chaperones and others.The mRNA expressions of two proteins - 14-3-3 protein sigma and Stress-induced-phosphoprotein 1 - were found to correlate with the corresponding proteins' abundance.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. saiful72@um.edu.my.

ABSTRACT

Background: A limiting factor in performing proteomics analysis on cancerous cells is the difficulty in obtaining sufficient amounts of starting material. Cell lines can be used as a simplified model system for studying changes that accompany tumorigenesis. This study used two-dimensional gel electrophoresis (2DE) to compare the whole cell proteome of oral cancer cell lines vs normal cells in an attempt to identify cancer associated proteins.

Results: Three primary cell cultures of normal cells with a limited lifespan without hTERT immortalization have been successfully established. 2DE was used to compare the whole cell proteome of these cells with that of three oral cancer cell lines. Twenty four protein spots were found to have changed in abundance. MALDI TOF/TOF was then used to determine the identity of these proteins. Identified proteins were classified into seven functional categories - structural proteins, enzymes, regulatory proteins, chaperones and others. IPA core analysis predicted that 18 proteins were related to cancer with involvements in hyperplasia, metastasis, invasion, growth and tumorigenesis. The mRNA expressions of two proteins - 14-3-3 protein sigma and Stress-induced-phosphoprotein 1 - were found to correlate with the corresponding proteins' abundance.

Conclusions: The outcome of this analysis demonstrated that a comparative study of whole cell proteome of cancer versus normal cell lines can be used to identify cancer associated proteins.

No MeSH data available.


Related in: MedlinePlus