MicroRNA-216a promotes the metastasis and epithelial–mesenchymal transition of ovarian cancer by suppressing the PTEN/AKT pathway

Figures & data

Figure 1 The expression levels of miR-216a in OC tissues and cells.

Notes: (A) The expression of miR-216a was compared between normal, benign, borderline, and OC tissues. (B) The expression of miR-216a was compared between 5 different OC cell lines (CAOV3, HO-8910, A2780, ES-2, and SKOV-3) and immortalized human fallopian tube epithelial cell line FTE187 cells. *P<0.05, **P<0.01.
Abbreviation: OC, ovarian cancer.

Figure 2 The prognostic value of miR-216a for OC patients.

Note: (A) Overall survival and (B) disease-free survival were compared between patients of high level of miR-216a and those of low level of miR-216a.
Abbreviation: OC, ovarian cancer.

Table 1 The clinical features of all included patients and the correlations between clinical features and miR-216a expression

Characteristics	Number of patients (n=87)	miR-520g expression		P-value
		Low group (n=43)	High group (n=44)
Age				0.500
≤50	29	16	13
>50	58	27	31
Ascites				0.644
≤100	27	12	15
>100	60	31	29
Serum CA-125				0.713
≤35	8	3	5
>35	79	40	39
Tumor size				0.101
≤3 cm	61	34	27
>3 cm	26	9	17
Lymph node metastasis				<0.01
Negative	49	34	15
Positive	38	9	29
FIGO stage				0.014
I–II	12	10	2
III–IV	75	33	42

Abbreviation: FIGO, Federation Internationale of Gynecologie and Obstetrigue.

Figure 3 MiR-216a promotes the migration and invasion of OC cells.

Notes: (A) MiR-216a mimics significantly elevated the expression level of miR-216a in CAOV3 cells. (B) Overexpression of miR-216a significantly increased the migration and invasion of CAOV3 cells. *P<0.05, **P<0.01. (C) MiR-216a inhibitor significantly decreased the expression level of miR-216a in SKOV-3 cells. **P<0.01. (D) Downregulation of miR-216a significantly reduced the migration and invasion of SKOV-3 cells. **P<0.01.
Abbreviations: OC, ovarian cancer; NC, negative control.

Figure 4 MiR-216a promotes the EMT phenotype of OC cells.

Notes: (A) Western blot results of E-cadherin and Vimentin in CAOV3 cells transfected with miR-216a mimics; **P<0.01. (B) Western blot results of E-cadherin and Vimentin in SKOV-3 cells transfected with miR-216a inhibitor; **P<0.01.
Abbreviations: EMT, epithelial–mesenchymal transition; OC, ovarian cancer; NC, negative control.

Figure 5 PTEN is a direct target of miR-216a in OC cells.

Notes: (A) MiR-216a and its complementary sequence in the 3′-UTR of PTEN. (B) Overexpression of miR-216a significantly inhibited the luciferase activity that carried wt 3′-UTR of PTEN but had no obvious influence on mt 3′-UTR of PTEN in CAOV3 cells. **P<0.01. (C) Inhibition of miR-216a obviously increased the luciferase activity that carried wt 3′-UTR of PTEN but had no obvious influence on mt 3′-UTR of PTEN in SKOV-3 cells. **P<0.01. (D) Overexpression of miR-216a significantly decreased PTEN mRNA level in CAOV3 cells. **P<0.01. (E) Inhibition of miR-216a significantly increased PTEN mRNA level in SKOV-3 cells. **P<0.01. (F) Overexpression of miR-216a significantly inhibited the expression of PTEN and increased p-AKT in CAOV3 cells. *P<0.05, **P<0.01. (G) Inhibition of miR-216a significantly increased the expression of PTEN and decreased p-AKT in SKOV-3 cells. *P<0.05, **P<0.01.
Abbreviations: OC, ovarian cancer; PTEN, phosphatase and tensin homolog; wt, wild type; mRNA, messenger RNA; NC, negative control; mt, mutant.

Figure 6 MiR-216a promotes the migration, invasion, and EMT of OC cells by suppressing PTEN/AKT pathway.

Notes: (A) Western blot analysis of the expression of PTEN, E-cadherin, Vimentin, p-AKT in CAOV3-miR-216a cells transfected with PTEN vector and control vector; *P<0.05, **P<0.01. (B) Restoration of PTEN in CAOV3-miR-216a cells abrogated the promoting effect of miR-216a mimics on migration and invasion. **P<0.01. (C) Western blot analysis of the expression of PTEN, E-cadherin, Vimentin, p-AKT in SKOV-3-anti-miR-216a cells transfected with PTEN siRNA and negative control vector; *P<0.05, **P<0.01. (D) Inhibition of PTEN in SKOV-3-anti-miR-216a cells reversed the inhibitory effect of miR-216a inhibitor on cell migration and invasion. **P<0.01.
Abbreviations: EMT, epithelial–mesenchymal transition; OC, ovarian cancer; PTEN, phosphatase and tensin homolog; NC, negative control.