PURPOSE: CD44 is overexpressed in various tumors including hepatocellular carcinoma (HCC). The purpose of this study was to examine the effects of CD44 antisense oligonucleotide (ASO) alone or combination with doxorubicin on HCC cells in vitro. METHODS: Cytotoxicity was measured by use of a cell viability assay in HCC cell line SNU-449. Tumorigenesis and invasion were accessed by colony formation, growth in soft agar and ECMatrix invasion assay. Apoptosis and necrosis were evaluated by using double staining with Hoechst 33342 and propidium iodide. Protein expression and mRNA level were detected by Western blot and RT-PCR. RESULTS: We have designed novel CD44 ASO, which can effectively down-regulate CD44 expression in SNU-449. Colony formation, growth in soft agar and invasion were significantly impaired after CD44 ASO treatment in SNU-499. In company with CD44 down-regulated by CD44 ASO, MDR-1 and Bcl-2 expression were also greatly reduced. CD44 ASO also increased chemosensitivity to doxorubicin significantly, lowered IC(50 )by one order of magnitude. Apoptosis and necrosis were also induced by CD44 ASO alone or in combination treatment with doxorubicin. CONCLUSIONS: Inhibition of CD44 expression by CD44 ASO significantly induced apoptosis, decreased tumorigenesis and invasion, and increased chemosensitivity. Thus, CD44 ASO is potentially a therapy that is worth investigating in the clinical setting.
PURPOSE:CD44 is overexpressed in various tumors including hepatocellular carcinoma (HCC). The purpose of this study was to examine the effects of CD44 antisense oligonucleotide (ASO) alone or combination with doxorubicin on HCC cells in vitro. METHODS:Cytotoxicity was measured by use of a cell viability assay in HCC cell line SNU-449. Tumorigenesis and invasion were accessed by colony formation, growth in soft agar and ECMatrix invasion assay. Apoptosis and necrosis were evaluated by using double staining with Hoechst 33342 and propidium iodide. Protein expression and mRNA level were detected by Western blot and RT-PCR. RESULTS: We have designed novel CD44ASO, which can effectively down-regulate CD44 expression in SNU-449. Colony formation, growth in soft agar and invasion were significantly impaired after CD44ASO treatment in SNU-499. In company with CD44 down-regulated by CD44ASO, MDR-1 and Bcl-2 expression were also greatly reduced. CD44ASO also increased chemosensitivity to doxorubicin significantly, lowered IC(50 )by one order of magnitude. Apoptosis and necrosis were also induced by CD44ASO alone or in combination treatment with doxorubicin. CONCLUSIONS: Inhibition of CD44 expression by CD44ASO significantly induced apoptosis, decreased tumorigenesis and invasion, and increased chemosensitivity. Thus, CD44ASO is potentially a therapy that is worth investigating in the clinical setting.
Authors: Karl E Miletti-González; Kyle Murphy; Muthu N Kumaran; Abhilash K Ravindranath; Roman P Wernyj; Swayamjot Kaur; Gregory D Miles; Elaine Lim; Rigel Chan; Marina Chekmareva; Debra S Heller; David Foran; Wenjin Chen; Michael Reiss; Elisa V Bandera; Kathleen Scotto; Lorna Rodríguez-Rodríguez Journal: J Biol Chem Date: 2012-03-20 Impact factor: 5.157
Authors: Jon C Henry; Jong-Kook Park; Jinmai Jiang; Ji Hye Kim; David M Nagorney; Lewis R Roberts; Soma Banerjee; Thomas D Schmittgen Journal: Biochem Biophys Res Commun Date: 2010-11-03 Impact factor: 3.575