Effects of hypoxia inducible factor-1alpha siRNA on the invasion of human Hela cells and expression of related proteins

Effects of hypoxia inducible factor-1alpha siRNA on the invasion of human Hela cells and expression of related proteins

Abstract

The effects of hypoxia on the invasion and the related protein expression of Hela cells and the role of hypoxia inducible factor-1α (HIF-1α) were investigated. The Hela cells were divided into three groups, namely, H₀ (non-transfected Hela cells), H₁ (pGenesil-1 empty plasmid-transfected Hela cells), and H₂ (HIF-1α-shRNA plasmid-transfected Hela cells), and were cultured under hypoxia (1% O₂) and normoxia for 48h. The expression of HIF-1α, E-cadherin, β-catenin, and actin was detected using Western blot. The scratch test and the invasion assay were applied to examine the invasion in each group. The expression of HIF-1α, E-cadherin, and β-catenin in tumor grafts was assayed immunohistochemically. Western blot results revealed that the bands of HIF-1α, E-cadherin, β-catenin, and actin proteins were detected in the H₀ and H₁ groups under hypoxia for 48h. The expression of E-cadherin, β-catenin, and actin was detected in the H₂ group under hypoxia for 48h, and normoxia. In the H₀, H₁, and H₂ groups under normoxia, and the H₂ group under hypoxia for 48h, no expression of HIF-1α was detectable. The scratch test showed that the invasive ability in the H₂ group was significantly alleviated. Immunohistochemically, it was found that there was a significant difference in the expression of HIF-1α, E-cadherin, and β-catenin between the H₁ and H₂ groups (P<0.05), but the difference was not significant between the H₀ and H₁ groups. It was concluded that the effects of hypoxia on the invasion of human cervical cancer Hela cells and the expression of related proteins (E-cadherin, β-catenin, and actin) depend on HIF-1α.

Keywords

hypoxia / actin;β-catenin / E-cadherin / invasion