The role of the retinoblastoma protein‑interacting zinc finger gene 1 tumor suppressor gene in human esophageal squamous cell carcinoma cells

Dong,Shangwen; Zhang,Peng; Liang,Shaojie; Wang,Shuo; Sun,Pei; Wang,Yuanguo

doi:10.3892/ol.2013.1608

The role of the retinoblastoma protein‑interacting zinc finger gene 1 tumor suppressor gene in human esophageal squamous cell carcinoma cells

Authors:
- Shangwen Dong
- Peng Zhang
- Shaojie Liang
- Shuo Wang
- Pei Sun
- Yuanguo Wang
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Affiliations: Department of Cardiothoracic Surgery, Tianjin Medical University General Hospital, Heping, Tianjin 300052, P.R. China, Tianjin Institute of Endocrinology, Tianjin Medical University, Heping, Tianjin 300070, P.R. China
Published online on: October 9, 2013 https://doi.org/10.3892/ol.2013.1608
Pages: 1656-1662

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Abstract

The tumor suppressor protein retinoblastoma protein‑interacting zinc finger gene 1 (RIZ1) is downregulated in several types of cancer, including esophageal squamous cell carcinoma (ESCC). The present study used two in vitro methods to re‑express RIZ1 in the human ESCC TE13 cell line in order to induce apoptosis. RIZ1 was re‑expressed in the TE13 cells by reintroducing the gene through transfection or by removal of transcriptional repression through treatment with a DNA methyltransferase (DNMT) inhibitor. To reintroduce the gene, the open reading frame of the RIZ1 gene was inserted into the eukaryotic expression pcDNA3.1(+) vector and pcDNA3.1(+)/RIZ1 was purified and transfected into the TE13 ESCC cells. Removing transcriptional repression involved treating the TE13 cells with 5‑aza‑2'‑deoxycytidine (5‑aza‑CdR), a DNMT inhibitor. RIZ1 mRNA and protein expression were determined by quantitative polymerase chain reaction (qPCR) and western blotting. The rate of apoptosis of the cells was determined by flow cytometry. A recombinant eukaryotic human RIZ1 expression plasmid, pcDNA3.1(+)/RIZ1, was constructed and confirmed by sequencing. RIZ1 mRNA and protein expression increased in pcDNA3.1(+)/RIZ1 stably transfected cells. Treatment with 5‑aza‑CdR for 48 and 72 h resulted in increased RIZ1 protein expression and increased the rate of apoptosis in the TE13 cells (P<0.01). In conclusion, transfection of the TE13 cells with the eukaryotic pcDNA3.1(+)/RIZ1 expression vector and reversal of transcriptional repression of RIZ1 using 5‑aza‑CdR demonstrate that it is possible to re‑express RIZ1 in TE13 cells. Furthermore, the re‑expression of RIZ1 led to an increased rate of apoptosis and this method may provide new therapeutic possibilities.

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