Effects of LncRNA SNHG14 on the biological behaviors of prostate cancer cells by targeting miR-449a/CCNE2
Objective To investigate the effects of long non coding RNA(LncRNA)small nucleolar RNA host gene 14(SNHG14)on the biological behaviors of prostate cancer(PCa)cells by targeting miR-449a/cyclin E2(CCNE2).Methods The expressions of LncRNA SNHG14,miR-449a,and CCNE2 mRNA in prostate cancer tissues and cancer cells were detected by qRT-PCR;The targeting relationship between LncRNA SNHG14 and miR-449a,and between CCNE2 and miR-449a were analyzed by dual luciferase assay;DU145 cells were divided into 5 groups including the control group,si-NC group,si-SNHG14 group,si-SNHG14+anti-NC group,si-SNHG14+anti miR-449a group.Except for the cells in the control group,the cells in all other groups were transfected with corresponding plasmids;Proliferation of DU 145 cells was measured by EdU assay;Apoptosis of DU145 cells was analyzed by flow cytometry;Migration of DU145 cells was evaluated by wound healing assay;Invasion of DU145 cells was examined by Transwell invasion assay;The expressions of Ki67,Cyclin D1,Bax,caspase-3,and CCNE2 proteins were detected by western blot;Subcutaneous tumor nude mouse model was used to assess the effects of LncRNA SNHG14 on the growth of tumors and miR-449a/CCNE2.Results LncRNA SNHG14 and CCNE2 were highly expressed in PCa tissues and cells,while miR-449a was lowly expressed(P＜0.05);Dual luciferase analyses showed there were targeted binding sites between LncRNA SNHG14 and miR-449a,and between CCNE2 and miR-449a;Compared with the si-NC group,the expression levels of LncRNA SNHG14,CCNE2,Cyclin D1 and Ki67,the positive rate of Edu,wound healing rate,and number of invaded cells in the si-SNHG14 group were all significantly decreased,whereas the expressions of miR-449a,Bax and caspase-3,cell apoptosis rate were all increased(P＜0.05);Compared with the si-SNHG14+anti-NC group,the expression levels of miR-449a,Bax and caspase-3,and cell apoptosis rate in the si-SNHG14+anti miR-449a group were all decreased,whereas the expression levels of CCNE2,Cyclin D1 and Ki67,positive rate of Edu,scratch healing rate,and number of invaded cells were increased.Analysis of subcutaneous tumors showed that the volume and weight of subcutaneous tumors in the si-SNHG14 group were smaller than those in the si-NC group,and they had lower expression in LncRNA SNHG14 and CCNE2,but a higher expression in miR-449a(P＜0.05).Conclusion Downregulation of LncRNA SNHG14 in PCa cells can inhibit proliferation,migration,invasion of cancer cells,and promote cancer cell apoptosis by regulating the miR-449a/CCNE2 signaling pathway.
NETL
NSTL
万方数据
