Research Paper Volume 11, Issue 22 pp 9982—9999

Circular RNA hsa_circ_0000515 acts as a miR-326 sponge to promote cervical cancer progression through up-regulation of ELK1

Figure 6. ELK1 targeted by miR-326 induces proliferation and invasion, and attenuates apoptosis and autophagy of cervical cancer cells. (A) the functional miR-326 binding sites in ELK1 3′UTR; (B) the relationship miR-326 and ELK1 analyzed by the dual luciferase reporter assay; (C) ELK1 expression in the normal adjacent tissues and the cervical cancer tissues determined by RT-qPCR; (D) Pearson correlation analysis of hsa_circ_0000515 and ELK1 expression; (E) Pearson correlation analysis of miR-326 and ELK1 expression; (F) ELK1 mRNA expression in cervical cancer cells determined by RT-qPCR; (G) ELK1 protein expression in cervical cancer cells determined by Western blot assay; (HI) EdU-stained cells (200 ×) and proliferation ability of Hela and SiHa cells assessed by EdU assay; (JK) cells invaded through the membrane in a Transwell system (200 ×) and the number of invaded Hela and SiHa cells; (LM) apoptosis ability evaluated by flow cytometry; (N) MDC staining showing autophagosome formation (400 ×) and quantitative analysis of the autophagosome number; (O) mRNA expression of PCNA, Caspase3, Caspase9, MMP-9, TIMP-1, Beclin1, P62, LC3-I and LC3-II determined by RT-qPCR; (Q) Western blots and protein expression of PCNA, Caspase3, Caspase9, MMP-9, TIMP-1, Beclin1, P62, LC3-I and LC3-II. *p < 0.05 vs. the NC group; #p < 0.05 vs. the normal adjacent group; &p < 0.05 vs. the OE-NC group; measurement data were expressed as mean ± standard deviation; unpaired t test was used to compare data with normal distribution and equal variance between cervical cancer (n = 63) and normal adjacent tissues (n = 40); one-way ANOVA was applied for comparison of data among multiple groups, followed by Tukey's post hoc test. The cell experiments repeated 3 times.