Effect of tumor suppressor gene cyclin‑dependent kinase inhibitor 2A wild‑type and A148T mutant on the cell cycle of human ovarian cancer cells

Bian,Zehua; Yu,Yang; Yang,Terigele; Quan,Chao; Sun,Wenjing; Fu,Songbin

doi:10.3892/ol.2014.1867

Effect of tumor suppressor gene cyclin‑dependent kinase inhibitor 2A wild‑type and A148T mutant on the cell cycle of human ovarian cancer cells

Authors:
- Zehua Bian
- Yang Yu
- Terigele Yang
- Chao Quan
- Wenjing Sun
- Songbin Fu
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Affiliations: Laboratory of Medical Genetics, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China
Published online on: February 11, 2014 https://doi.org/10.3892/ol.2014.1867
Pages: 1229-1232

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Abstract

Single‑base substitution may affect the function of genes. This study identified a single‑base substitution of G for A in codon 148 of cyclin‑dependent kinase inhibitor 2A (CDKN2A/p16) by sequencing human ovarian cancer cell line UACC‑1598. As a tumor suppressor gene, the expression of CDKN2A/p16 should be strictly controlled. In order to control CDKN2A/p16 gene expression, an inducible pTUNE vector system was selected. Using recombinant DNA technology, a CDKN2A/p16‑A148T and CDKN2A/p16‑wild‑type gene expression system was successfully constructed to investigate whether this single‑base substitution affects the function of CDKN2A/p16. For the wild‑type and the mutant, expression of CDKN2A/p16‑green fluorescent protein fusion protein increased markedly following isopropyl‑β‑D‑thiogalactoside induction, and was accompanied by significant G1 arrest in the transfected human ovarian cancer SKOV3 cell line. The inducible vectors used in this study, CDKN2A/p16‑wild‑type and CDKN2A/p16‑A148T open reading frame, may be useful for further investigation into whether this somatic mutation could alter the function of CDKN2A/p16 as a tumor suppressor gene. In summary, CDKN2A/p16‑A148T was identified in ovarian cancer cells, and this single‑base substitution did not affect the ability of CDKN2A/p16 to arrest the cell cycle.

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