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18 April 2016 : Laboratory Research

Effects of siRNA-Mediated Knockdown of HDAC1 on the Biological Behavior of Esophageal Carcinoma Cell Lines

Xing WangBC, Haisheng GuoCD, Weixin LiuDE, Chunmei YangFG, Lei YangABC, Dongguan WangDG, Xunguo WangAF

DOI: 10.12659/MSM.895853

Med Sci Monit 2016; 22:1291-1296

Abstract

BACKGROUND: HDAC1 has been shown to be closely associated with the occurrence of tumors. We aimed to investigate the effects of siRNA-mediated HDAC1 knockdown on the biological behavior of esophageal carcinoma cell lines.

MATERIAL AND METHODS: HDAC1 expression in esophageal cancer cell lines TE-1, Eca109, and EC9706 was compared by Western blot analysis. These cells were transfected with siRNA-HDAC1 and cell proliferation was evaluated by MTT assay to select the optimum cell line for subsequent experiments. The effects of siRNA-HDAC1 on the migration and invasion of the selected cell line were assessed by transwell assay. The expression of cell cycle-related proteins cyclinD1, p21 and p27, and epithelial-mesenchymal transition (EMT)-related protein zonula occludens-1 (ZO-1), E-cadherin and vimentin was determined by Western blot analysis.

RESULTS: HDAC1 expression in TE-1, Eca109 and EC9706 cells was significantly higher compared with normal esophageal cell line HEEC (P<0.01). MTT assay, Western blot and RT-PCR analyses demonstrated that the inhibitory effects of siRNA on HDAC1 expression and cell viability in TE-1 cells were the highest among all cell lines, which was therefore used in subsequent experiments. After TE-1 cells were transfected with siRNA-HDAC1, their migration and invasion were significantly lower compared with the controls (P<0.01). CyclinD1 and vimentin expression was significantly lower compared with the controls (P<0.01), whereas the expression of p21, p27, ZO-1 and E-cadherin was significantly higher (P<0.01).

CONCLUSIONS: The siRNA-mediated HDAC1 knockdown significantly inhibited the proliferation, migration and invasion of TE-1 cells probably by regulating the expression of cell cycle- and EMT-related proteins.

Keywords: Blotting, Western, Apoptosis - physiology, Carcinoma, Squamous Cell - pathology, Cell Cycle - physiology, Cell Movement - physiology, Cell Proliferation - physiology, Esophageal Neoplasms - pathology, Gene Knockdown Techniques, Histone Deacetylase 1 - metabolism, RNA, Messenger - metabolism, RNA, Small Interfering - genetics

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750