Promoter methylation of the DLC‑1 gene and its inhibitory effect on human colon cancer

Dai,Zhensheng; Jin,Yueling

doi:10.3892/or.2013.2551

Promoter methylation of the DLC‑1 gene and its inhibitory effect on human colon cancer

Authors:
- Zhensheng Dai
- Yueling Jin
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Affiliations: Department of Hematology‑Oncology, Shanghai Pudong Hospital Affiliated to Fudan University, Shanghai 201399, P.R. China, Department of Pathology, Shanghai Health Vocational and Technical College, Shanghai 200237, P.R. China
Published online on: June 19, 2013 https://doi.org/10.3892/or.2013.2551
Pages: 1511-1517

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Abstract

Deleted in liver cancer‑1 (DLC‑1), a candidate tumor suppressor gene which is inactive in liver carcinogenesis, is located at 8p21.3, where deletions are frequently found in several types of human cancer. Promoter hypermethylation is an epigenetic mechanism leading to silencing of the gene expression, which may be the primary cause for the absence of DLC‑1. We investigated the expression of the DLC‑1 gene and the methylation of the DLC‑1 gene in colon cancer cell lines (Caco‑2, LoVo and HT‑29). The data showed that reduced or undetectable levels of DLC‑1 mRNA were found in HT‑29 by reverse transcription-polymerase chain reaction (RT‑PCR). By contrast, the DLC‑1 gene was significantly expressed in Caco‑2 and LoVo cells. These findings were in agreement with the data obtained from western blot analysis. To further determine whether aberrant methylation is a contributing factor to transcriptional inactivation of DLC‑1 in HT‑29, the methylation of promoter was examined using methylation‑specific PCR and sodium bisulfite genomic sequencing in LoVo and HT‑29 cells, which suggests that promoter hypermethylation accounts for silencing of the DLC‑1 gene in HT‑29 cells. Since DLC‑1 is a candidate tumor suppressor gene, we sought to determine whether DLC‑1 expression is associated with cell proliferation in colon cancer cell lines. RNA interference techniques were adopted to inhibit DLC‑1 expression in the LoVo cell line and resulted in inhibition of cell growth and reduced colony formation. Collectively, our observations suggest that hypermethylation is responsible for abrogating the function of the DLC‑1 gene in colon cancer and indicate a role of DLC‑1 in colon carcinogenesis.

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