Xiaoling Zhu1, Wei Li2, Qinggang Meng2. 1. Department of Dermatology, The First Hospital of Harbin Harbin, Heilongjiang, China. 2. Department of Orthopaedic Surgery, The First Hospital of Harbin Harbin, Heilongjiang, China.
Abstract
OBJECTIVE: To determine the effects of H19 gene on the proliferation and invasion of A375 human melanoma cells. METHODS: A375 cells were infected with a lentiviral vector overexpressing H19 or transfected with H19 RNA interference constructs. The proliferation and invasion of A375/H19+ cells were measured by MTT cell viability and transwell assays, respectively. The mRNA and protein expression levels of matrix metalloproteinase 2 (MMP2) and MMP9 were measured by quantitative PCR (qPCR) and Western blotting, respectively, and the protein expression of Akt, phosphorylated Akt (p-Akt), Slug and E-cadherin were examined by Western blot analysis. RESULTS: The optical density value of A375/H19+ cells increased after incubation with MTT reagent for 12 h (P < 0.05), and the transwell assay showed that the average penetration rate of A375/H19+ cells significantly increased (P < 0.001). The expression levels of MMP2 and MMP9 were significantly increased in A375/H19+ cells, as determined by qPCR and Western blotting (P < 0.001). Moreover, A375/H19+ cells had upregulated levels of Slug and p-Akt and downregulated E-cadherin (P < 0.001). The weight and volume of A375/H19+ cell xenografts in nude mice were significantly increased, but its inhibition rate was smaller (P < 0.05 and P < 0.001, respectively). CONCLUSIONS: The results of this study showed that H19 overexpression promoted the proliferation, invasion, and growth of A375 cells. In addition, it upregulated the mRNA and protein expression levels of MMP2 and MMP9, which in turn promoted cell invasion. Furthermore, H19 appeared to enhance Akt phosphorylation, directly suppress E-cadherin, and upregulate Slug expression to promote A375 cell invasion. IJCEP
OBJECTIVE: To determine the effects of H19 gene on the proliferation and invasion of A375humanmelanoma cells. METHODS:A375 cells were infected with a lentiviral vector overexpressing H19 or transfected with H19 RNA interference constructs. The proliferation and invasion of A375/H19+ cells were measured by MTT cell viability and transwell assays, respectively. The mRNA and protein expression levels of matrix metalloproteinase 2 (MMP2) and MMP9 were measured by quantitative PCR (qPCR) and Western blotting, respectively, and the protein expression of Akt, phosphorylated Akt (p-Akt), Slug and E-cadherin were examined by Western blot analysis. RESULTS: The optical density value of A375/H19+ cells increased after incubation with MTT reagent for 12 h (P < 0.05), and the transwell assay showed that the average penetration rate of A375/H19+ cells significantly increased (P < 0.001). The expression levels of MMP2 and MMP9 were significantly increased in A375/H19+ cells, as determined by qPCR and Western blotting (P < 0.001). Moreover, A375/H19+ cells had upregulated levels of Slug and p-Akt and downregulated E-cadherin (P < 0.001). The weight and volume of A375/H19+ cell xenografts in nude mice were significantly increased, but its inhibition rate was smaller (P < 0.05 and P < 0.001, respectively). CONCLUSIONS: The results of this study showed that H19 overexpression promoted the proliferation, invasion, and growth of A375 cells. In addition, it upregulated the mRNA and protein expression levels of MMP2 and MMP9, which in turn promoted cell invasion. Furthermore, H19 appeared to enhance Akt phosphorylation, directly suppress E-cadherin, and upregulate Slug expression to promote A375 cell invasion. IJCEP
Authors: L Yan; J Zhou; Y Gao; S Ghazal; L Lu; S Bellone; Y Yang; N Liu; X Zhao; A D Santin; H Taylor; Y Huang Journal: Oncogene Date: 2014-08-04 Impact factor: 9.867
Authors: Jacques Ferlay; Isabelle Soerjomataram; Rajesh Dikshit; Sultan Eser; Colin Mathers; Marise Rebelo; Donald Maxwell Parkin; David Forman; Freddie Bray Journal: Int J Cancer Date: 2014-10-09 Impact factor: 7.396
Authors: Amanda N Kallen; Xiao-Bo Zhou; Jie Xu; Chong Qiao; Jing Ma; Lei Yan; Lingeng Lu; Chaochun Liu; Jae-Sung Yi; Haifeng Zhang; Wang Min; Anton M Bennett; Richard I Gregory; Ye Ding; Yingqun Huang Journal: Mol Cell Date: 2013-09-19 Impact factor: 17.970