Abstract
Osteosarcoma (OS) is a type of primary malignant bone tumor with a high propensity for local recurrence and distant metastasis. A previous study showed Snail-1 is highly expressed in OS cells. The present study aimed to investigate the association between the transcription factor Snai1 and E-cadherin in OS. SaOS2 OS cells were transfected either with a plasmid expressing short hairpin RNA (shRNA) specific for the Snai1-1 gene (SaOS2-shRNA) or a negative control plasmid (SaOS2-Mock). The expression levels of E-cadherin and Snai1-1 in the transfected and control cells were determined by quantitative polymerase chain reaction and western blot analysis. In addition, the study was extended to evaluate the migratory and invasive properties of the cells through a Transwell experiment. The results show that E-cadherin was expressed at a high level in the SaOS2-shRNA cells, which were much less migratory and invasive than the control cells. Overexpression of Snai1-1 in OS is associated with tumor progression, possibly through the suppression of E-cadherin expression and induction of the process of epithelial-mesenchymal transition, which contributes to the proceeding invasion and metastasis of OS cells.
Highlights
Materials and methodsOsteosarcoma (OS) is the most common type of primary malignancy of bone
To determine the association between Snail‐1 and E‐cadherin, short hairpin RNA (shRNA) targeting Snail‐1 was successfully transfected into SaOS2 cells and the expression levels of Snail‐1 and E‐cadherin in the cells were detected by western blot analysis
The expression levels of E‐cadherin in the SaOS2 cells transfected with the Snail‐1‐shRNA vector were significantly higher than those in the SaOS2 cells with no treatment or infected with the negative control shRNA (Fig. 1)
Summary
Osteosarcoma (OS) is the most common type of primary malignancy of bone. Despite intensive chemotherapy and adequate surgical resection, ~30‐50% of patients succumb to OS, mainly due to distant metastasis to the lung [1,2]. YANG et al: SNAI1-1 INDUCES OSTEOSARCOMA INVASION AND METASTASIS BY INHIBITING E-CADHERIN was stably suppressed and mock‐transfected cells, the SaOS2 cells were transfected with a plasmid (pcDNA3.1/shSnail‐1 or pcDNA3.1/GAPDH, respectively; Introgen Therapeutics Inc., Austin, TX, USA) for two days. The stable clones were selected by maintaining the cells in medium containing the antibiotic G418 (Nanjing KeyGen Biotech Co., Ltd., Nanjing, China). The cells were suspended at a density of 1x105 cells/ml in 500 ml McCoy's 5A medium supplemented with 0.5% FBS, and 1,25(OH)2‐D3 with a concentration of 10‐6 M was added to the 8‐mm porous BD BioCoat Matrigel chamber inserts (BD Biosciences, San Jose, CA, USA). Statistical comparisons were performed with the software package SPSS, version 13.0 (SPSS, Inc., Chicago, IL, USA) using Student's t‐test for paired observations or one‐way analysis of variance with Student‐Newman‐Keuls, least significant difference and Dunnett's methods. The mean values and SD were calculated for the experiments conducted in triplicate
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