Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer

Potchanapond Graidist, Raphatphon Nawakhanitworakul, Jiraporn Saekoo, Chavaboon Dechsukhum, Kenichi Fujise

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: WT1 was originally identified in Wilms tumor, a childhood kidney cancer. This gene was expressed in wide variety of solid cancers. Alternative splicing of WT1 transcript generates four major protein isoforms and thirty-six minor protein isoforms, each having different functional properties. WT1 gene has been considered as a tumor suppressor gene and anti-apoptotic protein. However, the mechanism of WT1 in breast cancer remains unclear. Objective: Evaluate the role of truncated WT1 isoforms (T-KTS+ and T-KTS-) and two major WT1 isoforms (+/+ and +/-) in apoptosis in breast cancer cell line, MCF-7. Materials and methods: RNA interference (RNAi) was employed in an attempt to define the role of WT1 in a breast cancer cell line (MCF-7). Furthermore, MCF-7 overe-xpressing cells that stably expressed two truncated WT1 isoforms (T-KTS+ and T-KTS-) or two major WT1 isoforms (+/+ and +/-) were generated and exposed to Doxorubicin. The mortality of cells was determined as a percentage of trypan blue-stained cells in total cells. The apoptotic molecules in apoptosis pathway were detected using RT-PCR, caspase-7 activity assay and Western blot analysis techniques. Results: Transfection of siRNAWT1 into MCF-7 cells resulted in decreasing of WT1 protein and related to the increasing in number of cell death and caspase-7 activity. Over-expression of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms protected cells from cell death induced by apoptosis-inducing agent, doxorubicin. Moreover, the expression of apoptotic p53, Bak and caspase-7 were decreased by the expression of all four WT1 isoforms, especially T-KTS- and T-KTS+ isoforms. Conclusion: T-KTS+ and T-KTS- isoforms as well as WT1+/+ and WT1+/- isoforms could function as an antiapoptotic protein in breast cancer cell line, MCF-7.

Original languageEnglish (US)
Pages (from-to)711-720
Number of pages10
JournalAsian Biomedicine
Volume4
Issue number5
StatePublished - Oct 2010

Fingerprint

Protein Isoforms
Breast Neoplasms
Caspase 7
Genes
Cells
Cell death
Apoptosis
Cell Line
Doxorubicin
Tumors
Cell Death
Apoptosis Regulatory Proteins
Trypan Blue
Wilms Tumor
Kidney Neoplasms
MCF-7 Cells
Alternative Splicing
RNA Interference
Tumor Suppressor Genes
Transfection

Keywords

  • Anti-apoptosis
  • MCF-7
  • Truncated WT1
  • WT1

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Graidist, P., Nawakhanitworakul, R., Saekoo, J., Dechsukhum, C., & Fujise, K. (2010). Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer. Asian Biomedicine, 4(5), 711-720.

Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer. / Graidist, Potchanapond; Nawakhanitworakul, Raphatphon; Saekoo, Jiraporn; Dechsukhum, Chavaboon; Fujise, Kenichi.

In: Asian Biomedicine, Vol. 4, No. 5, 10.2010, p. 711-720.

Research output: Contribution to journalArticle

Graidist, P, Nawakhanitworakul, R, Saekoo, J, Dechsukhum, C & Fujise, K 2010, 'Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer', Asian Biomedicine, vol. 4, no. 5, pp. 711-720.
Graidist P, Nawakhanitworakul R, Saekoo J, Dechsukhum C, Fujise K. Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer. Asian Biomedicine. 2010 Oct;4(5):711-720.
Graidist, Potchanapond ; Nawakhanitworakul, Raphatphon ; Saekoo, Jiraporn ; Dechsukhum, Chavaboon ; Fujise, Kenichi. / Anti-apoptotic function of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms in breast cancer. In: Asian Biomedicine. 2010 ; Vol. 4, No. 5. pp. 711-720.
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AB - Background: WT1 was originally identified in Wilms tumor, a childhood kidney cancer. This gene was expressed in wide variety of solid cancers. Alternative splicing of WT1 transcript generates four major protein isoforms and thirty-six minor protein isoforms, each having different functional properties. WT1 gene has been considered as a tumor suppressor gene and anti-apoptotic protein. However, the mechanism of WT1 in breast cancer remains unclear. Objective: Evaluate the role of truncated WT1 isoforms (T-KTS+ and T-KTS-) and two major WT1 isoforms (+/+ and +/-) in apoptosis in breast cancer cell line, MCF-7. Materials and methods: RNA interference (RNAi) was employed in an attempt to define the role of WT1 in a breast cancer cell line (MCF-7). Furthermore, MCF-7 overe-xpressing cells that stably expressed two truncated WT1 isoforms (T-KTS+ and T-KTS-) or two major WT1 isoforms (+/+ and +/-) were generated and exposed to Doxorubicin. The mortality of cells was determined as a percentage of trypan blue-stained cells in total cells. The apoptotic molecules in apoptosis pathway were detected using RT-PCR, caspase-7 activity assay and Western blot analysis techniques. Results: Transfection of siRNAWT1 into MCF-7 cells resulted in decreasing of WT1 protein and related to the increasing in number of cell death and caspase-7 activity. Over-expression of T-KTS+, T-KTS-, WT1+/+ and WT1+/- isoforms protected cells from cell death induced by apoptosis-inducing agent, doxorubicin. Moreover, the expression of apoptotic p53, Bak and caspase-7 were decreased by the expression of all four WT1 isoforms, especially T-KTS- and T-KTS+ isoforms. Conclusion: T-KTS+ and T-KTS- isoforms as well as WT1+/+ and WT1+/- isoforms could function as an antiapoptotic protein in breast cancer cell line, MCF-7.

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