Article Figures & Data
Figures
- 1
PVT1 and miR-214 expression in ovarian cancer cell lines and tissues. (A) The expression of PVT1 was significantly higher in tumor tissues compared to normal ovarian tissues. (B) The expression of miR-214 was obviously lower in tumor tissues compared to normal ovarian tissues. (C) The expression of PVT1 was up-regulated in ovarian cancer cell lines (SKOV3, HO8910, ES2, A2780, and SW626) compared to human primary ovarian cells and PVT1 expression was the highest in SKOV3 cells. (D) The expression of miR-214 was down-regulated in ovarian cancer cell lines (SKOV3, HO8910, ES2, A2780, and SW626) compared to human primary ovarian cells and miR-214 expression was the lowest in SKOV3 cells (*P<0.05).
- 2
Kaplan–Meier analysis for PFS and OS of EOC. (A) High PVT1 expression demonstrated the shorter median of PFS compared to low PVT1 expression (28 vs. 34 months, P=0.016). (B) High PVT1 expression demonstrated the shorter median of OS compared to low PVT1 expression (40 vs. 51 months, P=0.020). (C) Low miR-214 expression demonstrated the shorter median of PFS compared to high miR-214 expression (28 vs. 31 months, P=0.021). (D) Low miR-214 expression demonstrated the shorter median of OS compared to high miR-214 expression (41 vs. 44 months, P=0.034).
- 3
Knockdown of PVT1 inhibits ovarian cancer cell proliferation, migration, and invasion. (A) Satisfactory repressing efficiency was achieved at 48 h after transfection. (B) CCK-8 assay showed that knockdown of PVT1 inhibited SKOV3 cell proliferation. (C) Invasion assay showed that knockdown of PVT1 dramatically decreased SKOV3 cell invasion. (D) Wound healing assay showed that knockdown of PVT1 inhibited SKOV3 cell migration. (E) Mesenchymal marker proteins (Vimentin and β-catenin) and EMT-mediating transcription factor (Snail and Slug) were down-regulated in SKOV3 cells with PVT1-knockdown, while epithelial marker E-cadherin was up-regulated (*P<0.05).
- 4
PVT1 inhibits the miR-214 expression by interacting with EZH2 in ovarian cancer cells. (A) PVT1 was enriched in the anti-EZH2 RIP fraction compared to the anti-IgG fraction in SKOV3 cells. (B) After PVT1 was knocked down, miR-214 was up-regulated in SKOV3 cells. (C) MiR-214 was significantly up-regulated after si-EZH2 was transfected into SKOV3 cells. (D) Chromatin immunoprecipitation (ChIP) method proved that PVT1 silencing decreased the binding ability between EZH2 and miR-214 promoter compared to IgG group. (E) The expression of miR-214 was decreased in PVT1 repressing SKOV3 cells using miR-214 inhibitor. (F) CCK-8 assay showed that si-PVT1 inhibited the proliferation of SKOV3 cells and miR-214 inhibitor could counteract the si-PVT1 inhibiting effect of SKOV3 cells. (G) Wound healing assay showed that si-PVT1 inhibited the migration of SKOV3 cells. However, miR-214 inhibitor could counteract the si-PVT1 inhibiting effect in SKOV3 cells. (H) Invasion assay showed that si-PVT1 inhibited the invasion of SKOV3 cells. However, miR-214 inhibitor could counteract the si-PVT1 inhibiting effect in SKOV3 cells (*P<0.05).
Tables
Characteristics Case (n, %) Age at surgery (years) Media: 51; range: 28–76 ≤51 136 (58.9) >51 95 (41.1) Body mass index (kg/m2) 24.18±3.78 Menopausal status Yes 154 (66.7) No 77 (33.3) Pathologic type Serous 116 (50.2) Mucous and others* 115 (49.8) Histologic grade G1–2 141 (61.0) G3 90 (39.0) FIGO stage** I–II 81 (35.1) III–IV 150 (64.9) Lymph nodes metastasis No 186 (80.5) Yes 45 (19.5) Residual tumor (cm) <1 187 (81.0) ≥ 1 44 (19.0) Ascites volume (mL) Media: 1000; range: 200–7000 ≤1000 157 (68.0) >1000 74 (32.0) Serum CA125 (U/mL) Media: 675; range: 23–5400 ≤675 140 (60.6) >675 91 (39.4) *Others: others include the endometrioid, clear cell, and undifferentiated ovarian cancers.
**FIGO: Federation International of Gynecology and Obstetrics.
- 2
Correlation of PVT1 and miR-214 expression with clinicopathologic characteristics of EOC patients
Characteristics n PVT1 expression, n P MiR-214 expression, n P Low High Low High Age (years) 0.679 0.885 ≤51 136 61 75 76 60 >51 95 40 55 54 41 Menopause 0.302 0.453 Yes 154 71 83 84 70 No 77 30 47 46 31 Pathologic type 0.940 0.129 Serous 116 51 65 71 45 Mucous and others* 115 50 65 59 56 Histologic grade 0.924 <0.001 G1–2 141 62 79 63 78 G3 90 39 51 67 23 FIGO stage <0.001 <0.001 I–II 81 53 28 29 52 III–IV 150 48 102 101 49 LN metastasis 0.436 0.266 No 186 79 107 108 78 Yes 45 22 23 22 23 Residual tumor (cm) 0.552 0.204 <1 187 80 107 109 78 ≥ 1 44 21 23 21 23 Ascites volume (mL) 0.216 0.232 ≤1000 157 73 84 83 72 >1000 74 28 46 47 29 Serum CA125 (U/mL) 0.548 0.831 ≤675 140 59 81 78 62 >675 91 42 49 52 39 *Others: others include the endometrioid, clear cell, and undifferentiated ovarian cancers.
PVT1 expression, n (%) Total cases ρ P Low High MiR-214 expression 0.137 0.036 Low 49 (48.5) 81 (62.3) 130 High 52 (51.5) 49 (37.7) 101 Total cases 101 130 231 - 4
Univariate and multivariate survival analyses of the prognostic factors for overall and disease-free survival in EOC patients
Variable n Progression-free survival Overall survival Univariate analysis Multivariate analysis Univariate analysis Multivariate analysis Media of PFS Pa HR 95% CI For HR Pb Media of OS Pa HR 95% CI For HR Pb Age (years) 0.621 NA NA NA 0.676 NA NA NA ≤51 136 30 42 >51 95 31 44 Menopause 0.373 NA NA NA 0.488 NA NA NA Yes 154 30 43 No 77 32 44 Pathologic type 0.978 NA NA NA 0.849 NA NA NA Serous 116 30 41 Mucous and others* 115 31 44 Histologic grade 0.025 1.083 0.802–
1.4620.604 0.114 NA NA NA G1–2 141 42 44 G3 90 34 40 FIGO stage <0.001 2.265 1.632–
3.144<0.001 <0.001 1.994 1.423–
2.792<0.001 I–II 81 44 58 III –IV 150 24 39 LN metastasis 0.019 1.366 0.958–
1.9470.085 0.035 1.274 0.887–
1.8290.190 No 186 31 43 Yes 45 24 41 Residual tumor (cm) 0.277 NA NA NA 0.806 NA NA NA <1 187 31 43 ≥ 1 44 30 41 Ascites volume (mL) 0.237 0.403 NA NA NA ≤1000 157 40 NA NA NA 55 >1000 74 36 58 Serum CA125 (U/mL) 0.753 NA NA NA 0.991 NA NA NA ≤675 140 36 55 >675 91 39 55 PVT1 expression 0.016 1.111 0.819–
1.5060.498 0.020 1.153 0.842–
1.5790.375 Low 101 34 51 High 130 28 40 MiR-214 expression 0.021 1.246 0.925–
1.6800.148 0.034 1.193 0.880–
1.6160.256 Low 130 28 41 High 101 31 44 Pa: P value, log rank test; HR: hazard ratio; CI: confidence interval; Pb: P value, Cox regression; CR: complete response; PR: partial response; SD: table disease; PD: progressive disease; NA: not applicable. *Others: others include the endometrioid, clear cell, and undifferentiated ovarian cancers.
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