标题：
LincRNA-ROR induces epithelial-to-mesenchymal transition in high-grade ovarian serous cancer by increasing Wnt/β-catenin signaling
讲者：
娄艳辉
单位：
青岛大学医学院附属医院
播放：
1645
论文摘要：
Objective: To investigate the expression of long intergenic non-coding RNA-ROR (Linc-ROR) in high-grade ovarian serous cancer, to explore the relationship between Linc-ROR expression and biological function of high-grade ovarian serous cancer cells, and to analyze the effects of Linc-ROR on epithelial-to-mesenchymal transition(EMT) in high-grade ovarian serous cancer cells by increasing Wnt/β-catenin signaling pathway.
Methods: (1)We collected 34 high-grade ovarian serous cancer tissue samples, 20 normal ovarian tissue samples and 20 normal fallopian tube tissue samples between June 2014 and February 2016. Real-time reverse transcription (qRT)-PCR was used to detect the Linc-ROR expression in different samples. The relationship between Linc-ROR expression level and ovarian cancer International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis was analyzed. (2)Constructing Linc-ROR small interference RNA (siRNA) and pIRES2-EGFP-Linc-ROR plasmid, then Linc-ROR siRNA and pIRES2-EGFP-Linc-ROR plasmid were transfected into SKOV3 cells separately. Cell proliferation, migration and invasion ability was assessed by cell counting kit-8 (CCK-8), wound healing assay and transwell invasion assay. The protein expression of EMT related markers E-cadherin, β-catenin, vimentin and Wnt / β-catenin pathway target gene c-myc was detected by Western blot. (3)Ovarian cancer SKOV3 cells were treated with Lithium chloride (LiCl), cell proliferation was assessed using CCK-8 assay. The expression of E-cadherin, β-catenin, vimentin and c-myc protein was measured by Western blot. SKOV3 cells were treated with both LiCl and Linc-ROR siRNA, both LiCl and Linc-ROR siRNA-control, the expression of E-cadherin, β-catenin, vimentin and c-myc protein was measured by Western blot.
Results: (1)The expression level of Linc-ROR mRNA was significantly higher in high-grade ovarian serous cancer than normal ovarian tissues and normal fallopian tube tissues (4.38±2.55, 1.49±1.69, 1.45±1.57; F=8.62, P<0.01). With the progression of FIGO stages, the expression of Linc-ROR was increased (F=95.702, P<0.01), and it was associated with lymph node metastasis (t=7.397, P<0.01). (2)The expression of Linc-ROR mRNA in Linc-ROR siRNA-1 group, siRNA-2 group, siRNA-3 group cells was significantly lower than that in the Linc-ROR siRNA-NC group (siRNA-1：0.356±0.112，siRNA-2：0.760±0.137，siRNA-3：0.838±0.142，siRNA-NC：0.997±0.249; F=26.29, P<0.05). Compared with the negative control siRNA-NC group, the proliferation, migration and invasion ability of SKOV3 cells in siRNA-1 group were significantly decreased (P<0.01), the expression of E-cadherin protein was significantly increased, and the expression of β-catenin, vimentin and c-myc protein was significantly decreased (P<0.01). (3)The expression level of Linc-ROR mRNA in ROR group was significantly higher than that in Vector group (11.698±5.34、1.043±0.235; t= 6.304, P<0.01). Compared with the negative control Vector group, the proliferation, migration and invasion ability of SKOV3 cells in ROR group were significantly increased (P<0.01), the expression of E-cadherin protein was significantly decreased, and the expression of β-catenin, vimentin and c-myc protein was significantly increased (P<0.01). (4)The cell proliferation was significantly different in SKOV3 cells treated with different concentration of LiCl at different times (P<0.05), and the best concentration is 10mmol/L, the optimal time is 24 hours. Added LiCl to SKOV3 cells induced the decrease of E-cadherin protein expression (P<0.01), the increase of β-catenin, vimentin and c-myc protein expression (P<0.01). Compared with the LiCl group and LiCl + siRNA-control group, in LiCl + Linc-ROR siRNA group, the expression of E-cadherin protein was significantly up-regulated, and the expression of β-catenin, vimentin and c-myc protein was significantly down-regulated (P<0.01).
Conclusion: Linc-ROR is highly expressed in high-grade ovarian serous cancer and is closely related to FIGO stage and lymph node metastasis of high-grade ovarian serous cancer. Exogenous interference Linc-ROR expression could affect the proliferation, migration and invasion ability and EMT of high-grade ovarian serous cancer cells, which may play a role in regulating Wnt / β-catenin signaling pathway. Linc-ROR may be one of the important molecules in the occurrence and development, invasion and metastasis of high-grade ovarian serous cancer.
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