Title: Interaction between hypoxia-inducible factor-1α and Wnt/β-catenin signaling pathway in gastric cancer cells under hypoxic condition
Abstract: Objective To investigate the interaction between hypoxia-inducible factor-1α(HIF-1α) and Wnt/betacatenin signaling pathway in gastric cancer SGC-7901 cells under hypoxic condition.Methods SGC-7901 cells were transfected with plasmid β-catenin-micRNA,from which stably transfected cell strain SGC-7901-β-catenin-micRNA was screened.SGC-7901 cells were divided into three groups.The SGC-7901 cells in normal control group were cultured under normal condition for 48 h,while those in liposome group were cultured under normal condition for 24 h then added with 10 μl liposome and further cultured for 24 h,and those in negative control group were cultured under normal condition for 24 h then transfected with negative control plasmid and further cultured for 24 h.However,SGC-7901-β-cateninmicRNA cells were served as interference group and cultured under normal condition for 48 h.The doubling time,colony number on plate and cell cycle of cells in various groups were determined by doubling time test,plate colony formation test and flow cytometry respectively.In addition,SGC-7901 and SGC-7901-β-catenin-micRNA cells were divided into control,hypoxia and double hypoxia groups,separately.The cells in control groups were cultured under normal condition for 48 h,while those in hypoxia groups were cultured under normal condition for 32 h then under physically hypoxic condition for 16 h,and those in double hypoxia groups under normal condtion for 24 h,chemically hypoxic condition for 8 h, and physically hypoxic condition for 16 h.The expressions of HIF-1α and β-catenin at mRNA and protein levels in cells of various groups were determined by RT-PCR and Western blot respectively.Results Compared with those of SGC-7901 cells in normal control,liposome and negative control groups,the doubling time of SGC-7901-β-catenin-micRNA cells increased,and the mean colony number on plate decreased,while the percentage at G1 phase increased and that at S phase decreased(each P 0.05).Both the expression levels of HIF-1α and β-catenin mRNAs and proteins were significantly higher in SGC-7901 cells of hypoxia group than in those of control group,in SGC-7901 cells of double hypoxia group than in those of hypoxia group,in SGC-7901-β-catenin-micRNA cells of hypoxia group than in those of control group,and in SGC-7901-β-catenin-micRNA cells of double hypoxia group than in those of hypoxia group(each P 0.05).However, both the expression levels of HIF-1α and β-catenin mRNAs and proteins in SGC-7901-β-catenin-micRNA cells of hypoxia and double hypoxia groups were significantly lower than those in SGC-7901 cells(each P 0.05).Conclusion Interaction between HIF-1α and Wnt/β-catenine was observed.
Publication Year: 2013
Publication Date: 2013-01-01
Language: en
Type: article
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