IL-37通过调控LINC01554/miR-223-3p轴抑制卵巢癌细胞增殖、迁移和侵袭

IL-37 inhibits proliferation,migration and invasion of ovarian cancer cells by regulating LINC01554/miR-223-3p axis

目的:探讨IL-37对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响及作用机制。方法:RT-qPCR检测33例卵巢癌组织和癌旁正常组织LINC01554和miR-223-3p表达,Pearson相关性分析分析卵巢癌组织LINC01554和miR-223-3p表达的相关性。体外培养卵巢癌细胞Caov-3,经10、50、100 ng/ml IL-37干预后,CCK-8和克隆形成实验检测细胞增殖,划痕实验检测细胞迁移,Transwell检测细胞侵袭,Western blot检测细胞E-cadherin和N-cadherin蛋白表达,RT-qPCR检测细胞LINC01554和miR-223-3p表达。分别转染LINC01554过表达载体、LINC01554小干扰RNA或miR-223-3p模拟物至Caov-3细胞,上述方法观察过表达LINC01554、100 ng/ml IL-37对敲减LINC01554或过表达miR-223-3p的Caov-3细胞增殖、迁移和侵袭的影响。双荧光素酶报告基因实验验证LINC01554和miR-223-3p的靶向关系。结果:卵巢癌组织中LINC01554表达低于癌旁正常组织(P<0.05),而miR-223-3p表达高于癌旁组织(P<0.05)。Pearson相关性分析显示,卵巢癌组织LINC01554与miR-223-3p表达呈负相关(r=−0.9772,P<0.05)。Caov-3细胞经50、100 ng/ml IL-37干预后,细胞OD值、集落形成数、划痕愈合率、侵袭数及N-cadherin蛋白表达降低(P<0.05),而E-cadherin蛋白表达升高(P<0.05)。50、100 ng/ml IL-37促进Caov-3细胞LINC01554表达(P<0.05),而抑制miR-223-3p表达。过表达LINC01554降低Caov-3细胞OD值、集落形成数、划痕愈合率、侵袭数及N-cadherin蛋白表达(P<0.05),而促进E-cadherin蛋白表达。LINC01554可靶向结合miR-223-3p,且过表达LINC01554促进Caov-3细胞miR-223-3p表达。敲减LINC01554或过表达miR-223-3p均可减轻IL-37对Caov-3细胞增殖、迁移和侵袭的抑制作用。结论:IL-37可能通过调控LINC01554/miR-223-3p轴抑制卵巢癌细胞Caov-3增殖、迁移和侵袭。

Objective:To investigate effects and mechanism of IL-37 on proliferation,apoptosis,migration and invasion of ovarian cancer cells.Methods:RT-qPCR was used to detect expressions of LINC01554 and miR-223-3p in 33 cases of ovarian cancer tissues and adjacent normal tissues.Pearson correlation was used to analyze correlation between expressions of LINC01554 and miR223-3p in ovarian cancer tissues.Caov-3 ovarian cancer cells were cultured in vitro and treated with different doses of IL-37(10,50,100 ng/ml),CCK-8 and clone formation test were used to assess cell proliferation;cell migration and invasion were detected by scratch test and Transwell,respectively.Protein expressions of E-cadherin and N-cadherin in cells were detected by Western blot,and expressions of LINC01554 and miR-223-3p in cells were detected by RT-qPCR.LINC01554 overexpression vector,small interfering RNA against LINC01554 or miR-223-3p mimic were transfected into Caov-3 cells,and the above above methods were used to explore effects of LINC01554 overexpression and 100 ng/ml IL-37 on proliferation,migration and invasion of Caov-3 cells,or effects of 100 ng/ml IL-37 on proliferation,migration and invasion of Caov-3 cells with LINC01554 knockdown or miR-223-3p overexpression.Dual luciferase reporter experiment to verify targeting relationship between LINC01554 and miR-223-3p.Results:Expression of LINC01554 in ovarian cancer tissue was lower than that in adjacent normal tissues(P<0.05),while expression of miR-223-3p was higher than that in adjacent normal tissues(P<0.05).Pearson correlation analysis showed that expression of LINC01554 was negatively correlated with miR223-3p expression in ovarian cancer tissue(r=−0.9772,P<0.05).After Caov-3 cells were treated with 50 or 100 ng/ml IL-37,cell OD value,number of colonies,scratch healing rate,number of invasion cells and protein expression of N-cadherin were decreased(P<0.05),while protein expression of E-cadherin was increased(P<0.05).50,100 ng/ml IL-37 treatment promoted expression of LINC01554 in Caov-3 cells(P<0.05),while inhibited expression of miR-223-3p.Overexpression of LINC01554 suppressed cell viability,proliferation,migration and invasion of Caov-3 cells and reduced protein expression of N-cadherin(P<0.05),while elevated protein expression of E-cadherin(P<0.05).LINC01554 could targeting bind to miR-223-3p,and overexpression of LINC01554 promoted expression of miR-223-3p in Caov-3 cells.Knockdown of LINC01554 or overexpression of miR-223-3p reduced inhibitory effects of IL-37 on proliferation,migration and invasion of Caov-3 cells.Conclusion:IL-37 may inhibit proliferation,migration and invasion of ovarian cancer cells Caov-3 by regulating LINC01554/miR-223-3p axis.