circ_0086414靶向miR-155-5p抑制Hippo/YAP信号通路调控口腔鳞癌细胞增殖迁移和侵袭  

circ_0086414 Targeting miR-155-5p Inhibits Hippo/YAP Signaling Pathway to Regulate Proliferation Migration and Invasion of Oral Squamous Cell Carcinoma Cells

作　　者：李玲 宋青青 毕克 LI Ling;SONG Qingqing;BI Ke(Jinan People's Hospital,Shandong Jinan 271199,China)

机构地区：[1]山东省济南市人民医院口腔科,山东济南271199

出　　处：《河北医学》2023年第3期358-364,共7页Hebei Medicine

基　　金：山东省优秀中青年科学家科研奖励基金,(编号:BS2020SW817)。

摘　　要：目的:探究circ-0086414靶向miR-155-5p抑制Hippo/Yap信号通路调控口腔鳞癌细胞增殖、迁移和侵袭的影响。方法:RT-PCR检测HIOEC、Tca8113、CAL27、SCC-9细胞中circ-0086414表达。将未转染任何质粒的SCC-9细胞设为Control组;将分别转染pcDNA-circ-0086414、pcDNA-NC、miR-155-5p inhibitor、miR-NC inhibito质粒的SCC-9细胞中,并设为pcDNA-circ-0086414组、pcDNA-NC组、miR-155-5p组、miR-NC组;将pcDNA-circ-0086414质粒转染于SCC-9细胞中,同时在培养基中加入5μmoL/L Verteporfin共同培养,设为Verteporfin组。RT-PCR检测细胞中circ-0086414、miR-155-5p表达;CCK-8检测细胞增殖能力;Transwell小室法检测细胞侵袭能力;划痕实验检测细胞迁移能力;蛋白质印迹检测细胞中Lats1、p-Lats1、YAP、p-YAP蛋白表达;双荧光素酶报告系统实验验证circ-0086414和miR-155-5p的靶向关系。结果:和人永生化口腔上皮细胞HIOECcirc-0086414(1.00±0.10)、miR-155-5p(1.00±0.09)相比,口腔鳞癌细胞株Tca8113、CAL27、SCC-9细胞中circ-0086414(0.73±0.07)、(0.51±0.05)、(0.36±0.03)表达明显降低,miR-155-5p(1.36±0.11)、(1.57±0.13)、(1.89±0.15)表达明显升高(P<0.05);且SCC-9细胞中circ-0086414表达和miR-155-5p表达变化最显著(P<0.05)。和Control组相比,pcDNA-circ-0086414组细胞中circ-0086414表达(1.26±0.12)明显升高,miR-155-5p组细胞中miR-155-5p表达(0.73±0.07)明显降低(P<0.001)。和Control组相比,pcDNA-circ-0086414组细胞增殖、侵袭(123.56±5.87)和迁移(16.90±1.62)能力均明显降低(P<0.05);和pcDNA-circ-0086414组相比,Verteporfin组细胞增殖、侵袭和迁移能力明显增加(P<0.05)。和Control组相比,pcDNA-circ-0086414组细胞中Last1(1.12±0.10)、p-Last1(1.49±0.12)、p-YAP(1.41±0.11)蛋白表达明显增加,YAP蛋白(0.45±0.04)表达明显降低(P<0.001);和pcDNA-circ-0086414组相比,Verteporfin组细胞中Last1(0.61±0.06)、p-Last1(0.82±0.08)、p-YAP蛋白(0.58±0.05)表达明显降低,YAP蛋白(0.93±0.09)表达明显升高(P<0.00Objective:To investigate the effect of circ-0086414 targeting miR-155-5p on the inhibition of Hippo/Yap signaling pathway in regulating the proliferation,migration and invasion of oral squamous cell carcinoma cells.Methods:RT-PCR was used to detect the expression of circ-0086414 in HIOEC,Tca8113,CAL27 and SCC-9 cells.SCC-9 cells which were not transfected with any plasmid were set as control group;SCC-9 cells transfected with pcDNA-circ-0086414,pcDNA-NC,miR-155-5p inhibitor and miR-NC inhibitor plasmids were set as pcDNA-circ-0086414 group,pcDNA-NC group,miR-155-5p group and miR-NC group respectively;the pcDNA-circ-0086414 plasmid was transfected into SCC-9 cells,and 5μmol/L Verteporfin was co-cultured and set as Verteporfin group.The expression of circ-0086414 and miR-155-5p was detected by RT-PCR;CCK-8 was used to detect cell proliferation;Transwell chamber method was used to detect the invasive ability of cells;the cell migration ability was detected by scratch test;the expression of Lats1,p-Lats1,YAP,p-YAP protein in cells was detected by Western blot;the target relationship between circ-0,086,414 and miR-155-5p was verified by double luciferase reporting system experiment.Results:Compared with human immortalized oral epithelial cells HIOECcirc-0086414(1.00±0.10)and miR-155-5p(1.00±0.09),the expression of circ-0086414(0.73±0.07),(0.51±0.05)and(0.36±0.03)in oral squamous cell carcinoma cell lines Tca8113,CAL27 and SCC-9 significantly decreased,and the expression of miR-155-5p(1.36±0.11),(1.57±0.13)and(1.89±0.15)significantly increased(P<0.05);the expression of circ-0086414 and miR-155-5p in SCC-9 cells changed most significantly(P<0.05).Compared with the control group,the expression of circ-0086414 in pcDNA-circ-0086414 group was significantly higher(1.26±0.12),and the expression of miR-155-5p in miR-155-5p group was significantly lower(0.73±0.07)(P<0.001).Compared with the control group,the proliferation,invasion(123.56±5.87)and migration(16.90±1.62)of cells in pcDNA-circ-0086414 group significan

关 键 词：circ-0086414 miR-155-5p Hippo/Yap信号通路 口腔鳞癌 增殖 侵袭 迁移 

分 类 号：R739.8[医药卫生—肿瘤]