Altered expression of PTCH and HHIP in gastric cancer through their gene promoter methylation: Novel targets for gastric cancer

Song,Yu; Tian,Ye; Zuo,Yun; Tu,Jian-Cheng; Feng,Yu-Fang; Qu,Chen-Jiang

doi:10.3892/mmr.2013.1333

Altered expression of PTCH and HHIP in gastric cancer through their gene promoter methylation: Novel targets for gastric cancer

Authors:
- Yu Song
- Ye Tian
- Yun Zuo
- Jian-Cheng Tu
- Yu-Fang Feng
- Chen-Jiang Qu
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Affiliations: Department of Radiotherapy and Oncology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, P.R. China, Department of Radiotherapy and Oncology, The First People's Hospital of Zhangjiagang, Zhangjiagang, Jiangsu, P.R. China
Published online on: February 21, 2013 https://doi.org/10.3892/mmr.2013.1333
Pages: 1159-1168

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Abstract

Human hedgehog-interacting protein (HHIP) and protein patched homolog (PTCH) are two negative regulators of the hedgehog signal, however, the mechanism of action in gastric cancer is unknown. Methylation of TSG promoters has been considered as a causative mechanism of tumorigenesis. In the present study, we first determined the expression of PTCH and HHIP mRNA and protein in gastric cancer tissues and adjacent normal tissues, and then detected methylation of the two genes to associate their expression and gene promoter methylation in gastric cancer. Expression in gastric cancer tissues and the cancer cells (AGS) were evaluated by reverse transcription-PCR (RT-PCR), qRT-PCR and IHC, while the methylation expression was valued by methylation-specific PCR (MSP) and bisulfite sequencing PCR (BSP). Cell viability and apoptosis were analyzed by MTT assay and flow cytometry following treatment with 5-aza-dc. Results showed that PTCH and HHIP expression was reduced in gastric cancer tissues that were not associated with clinical features. Moreover, methylation of the promoters was reversely correlated with the expression. Following treatment with 5-aza-dc, AGS reduced cell viability and induced apoptosis, which is associated with upregulation of HHIP expression. The data demonstrated that loss of expression of HHIP and PTCH is associated with the methylation of gene promoters. In addition, 5-aza-dc-induced apoptosis correlated with the upregulation of HHIP expression in AGS. The findings demonstrated that the PTCH and HHIP genes may be novel targets for the control of gastric cancer.

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