Fig 1: SRCIN1 is a direct target gene of miR-665 in ovarian cancer cells. (A) Transfection of miR-665 mimic increased miR-665 expression in SKOV3 and ES2 cells. (B) miR-665 mimic reduced the relative luciferase activity of SKOV3 cells transfected with SRCIN1 3'UTR-WT compared with SRCIN1 3'UTR-Mut. (C) miR-665 mimic reduced relative luciferase activity in ES2 cells transfected with SRCIN1 3'UTR-WT compared with SRCIN1 3'UTR-Mut. **P<0.01; ***P<0.001. miR-665, microRNA-665; miR-NC, microRNA negative control; SRCIN1, Src kinase signaling inhibitor 1; UTR, untranslated region; WT, wild-type; Mut, mutant.
Fig 2: miR-665 regulates cell proliferation mainly through SRCIN1 in ovarian cancer cells. (A) In SKOV3 cells, transfection of SRCIN1 siRNA decreased SRCIN1 protein expression. (B) Quantitative analysis of SRCIN1 expression following transfection with SRCIN1 siRNA. ***P<0.001 vs. control siRNA. (C) In SKOV3 cells, transfection of miR-665 inhibitor increased the protein level of SRCIN1, which was downregulated after transfection with SRCIN1 siRNA. (D) Quantitative analysis of SRCIN1 expression following multiple transfections. (E) In SKOV3 cells, transfection of miR-665 inhibitor inhibited cell proliferation, which was reversed after transfection SRCIN1 siRNA. (F) In SKOV3 cells, transfection of miR-665 inhibitor inhibited colony formation, which was reversed after transfection with SRCIN1 siRNA. **P<0.01 vs. miR-NC inhibitor + control siRNA; ***P<0.001 vs. miR-NC inhibitor + control siRNA, control siRNA; #P<0.05 vs. miR-665 inhibitor + SRCIN1 siRNA; ###P<0.001 vs. miR-665 inhibitor + SRCIN1 siRNA. miR-665, microRNA-665; miR-NC, microRNA negative control; siRNA, small interfering RNA; SRCIN1, Src kinase signaling inhibitor 1.
Fig 3: SRCIN1 expression is negatively correlated with miR-665 levels in tumor tissues from patients with ovarian cancer. (A) Reverse transcription-quantitative PCR showed that the expression level of SCRIN1 was downregulated in tumor tissues compared with normal tissues from 40 patients with ovarian cancer. (B) Pearson's correlation analysis showed that SRCIN1 mRNA levels were negatively correlated with miR-665 levels in tumor tissues from 40 patients with ovarian cancer. ***P<0.001 vs. normal tissues. miR-665, microRNA-665; SRCIN1, Src kinase signaling inhibitor 1.
Fig 4: Downregulation of miR-665 increases SRCIN1 expression in ovarian cancer cells. (A) Binding sequence between SRCIN1 and miR-665 is presented. (B) Downregulation of miR-665 increased the protein level of SRCIN1 in SKOV3 and ES2 cells. (C) Quantitative analysis of SRCIN1 expression. (D) Reverse transcription-quantitative PCR showed that the miR-665 inhibitor increased the mRNA expression level of SRCIN1 in SKOV3 and ES2 cells. ***P<0.001. miR-665, microRNA-665; miR-NC, microRNA negative control; SRCIN1, Src kinase signaling inhibitor 1; UTR, untranslated region; Mut, mutant.
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