Effects Of Collagen Triple Helix Repeat Containing 1 (CTHRC1) Silencing On Epithelial–Mesenchymal Transition And Cellular Migration In Glioblastoma Cells

Objective: Observe the epithelial–mesenchymal transition(EMT)and cell migration of glioblastoma following CTHRC1 knockdown by the RNA interference technique.These data might provide scientific information for prognosis prediction and targeted therapy for glioblastoma.Methods: 1.Collecte 46 glioblastoma tissue specimens from patients and 16 normal tissue specimens from the normal cortex of patients without glioblastoma.We examine CTHRC1 m RNA and protein expression in both glioblastoma and normal cortex tissue and cells by quantitative reverse transcription polymerase chain reaction(q RTPCR)and western blot analysis.2.We purchase of human malignant glioma cell lines U-118MG、U-373 MG 、 U-87 MG and the normal human astrocyte cell line 1800.We examine CTHRC1 m RNA and protein expression in the four cell lines by quantitative reverse transcription polymerase chain reaction(q RT-PCR)and western blot analysis.3.U-87 MG cells were divided into three groups: the CTHRC1-si RNA group(treated with CTHRC1-si RNA and Lipofectamine? 2000),the vector group(treated with control si RNA and Lipofectamine? 2000),and the control group(treated with Lipofectamine? 2000 only).We examine CTHRC1 m RNA and protein expression in the three groups by quantitative reverse transcription polymerase chain reaction(q RT-PCR)and western blot analysis.4.We determined the effect of CTHRC1 on U-87 MG cell proliferation and transfection via an MTT assay and Transwell experiment.5.We examine the expression of E-cadherin 、N-cadherin,Snail,and Slug in the three groups by quantitative reverse transcription polymerase chain reaction(q RT-PCR)and western blot analysis.6.Examine the expression of β-catenin in the three groups by western blot analysis.Results: 1.The m RNA and protein expression levels of CTHRC1 in glioblastoma tissues were significantly higher than in normal cortex tissues.2.CTHRC1 m RNA and protein expression levels were higher in the glioblastoma cells U-118 MG,U-373 MG and U-87 MG than in the normal human astrocyte cell line 1800.3.The mRNA and protein expression levels of CTHRC1 were significantly decreased in the CTHRC1-si RNA-transfected group.There was no significant difference in the expression level of CTHRC1 m RNA and protein between the vector group and the control groups.4.The proliferation of U-87 MG cells was inhibited by CTHRC1-si RNA in a time-dependent manner,the CTHRC1-si RNA transfected group grew more slowly than the control group and vector group.The U-87 MG cell migration was significantly reduced by CTHRC1-si RNA according to the migration assay.There was no significant difference in U-87 MG cell migration rate between the vector group and the control group.5.Western blot and q RT-PCR results revealed increased protein and m RNA levels of E-cadherin and decreased protein and m RNA levels of N-cadherin,Snail,and Slug in the CTHRC1-si RNAtransfected group.There was no significant difference in the expression level of E-cadherin,N-cadherin,Snail,or Slug m RNA and protein level between the vector group and the control group.6.We observed that the protein expression of β-catenin was strongly downregulated in the CTHRC1-si RNA group.Conclusions: Compared with normal cortical tissue,CTHRC1 is overexpression in glioblastoma tissues.These results indicate that CTHRC1 inhibits glioblastoma cell migration by suppressing epithelial–mesenchymal transition(EMT).CTHRC1 has a potential cancerogenic role in glioblastoma.CTHRC1 might play an important role in the development of glioblastoma.