Study of metabotropic Glutamate receptor 5 silence on biological behaviors of ovarian cancer SKOV3 cells and possible mechanism

CHU Guangmin; ZHANG Jianbo; SUN Miaomiao

doi:10.3969/j.issn.1009-0460.2017.08.003

Abstract：Objective To explore the effect of small interfering RNA (siRNA) targeting metabotropic Glutamate receptor 5 (mGluR5)on the biological behaviors of ovarian cancer SKOV3 cells as well as possible mechanism.Methods siRNA designed for specific targeting mGluR5 were transfected into SKOV3 cells as transfection group.Meanwhile,antisense sequence was used as negative control (NC).48h after transfection,QPCR was used to confirm the interfering efficacy to ensure that siRNA can be used for following experiments.CCK-8 assay,EdU cell immunofluorescence staining assay,Hoechst 33342 staining assay,wound healing assay were used to detect the cytoactive,proliferation,apoptosis and migration of SKOV3 cells after transfection.The related protein of PTEN/Akt signaling pathway was evaluated by Westem blotting.Results The level of mGluR5 mRNA in transfection group was (18.3 ± 2.3) ％,significantly decreased compared with NC (P＜0.05).72h after transfection,the absorbance of transfection group was 2.4±0.3,lower than 4.1±0.2 of NC (P＜0.05).48h after transfection,the proliferative rate of SKOV3 cells was (9.2± 1.2)％ in transfection group,lower than (22.4±2.3) ％ of NC (P＜0.05);the cell apoptotic rate of transfection group was (28.2±2.2) ％,higher than (6.2± 1.3) ％ of NC (P＜0.05).In wound healing assay,relative wound closure rates of NC and transfection group were (100.0±2.1)％ and (48.6±4.3)％ (P＜0.05).Western blotting showed that p-Akt expression decreased,and PTEN expression increased after transfecion (P＜0.05),but the expression of Akt did not change obviously (P＞0.05).Conclusion Silencing the expression of mGluR5 can inhibit the proliferation,migration and enhance apoptosis of SKOV3 cells.PTEN/Akt signaling pathway may be involved in this process.