circDDX17靶向miR-224-3p对宫颈癌SiHa细胞放射敏感性的影响

Effect of circDDX17 targeting miR-224-3p on radiosensitivity of cervical cancer SiHa cells

目的探讨circDDX17对宫颈癌SiHa细胞放射敏感性的影响及可能机制。方法收集2018年8月至2020年8月于保定市第二医院行手术治疗的41例宫颈癌患者作为研究对象。对其宫颈癌组织和癌旁组织采用实时荧光定量PCR(qRT-PCR)法检测组织中circDDX17和miR-224-3p的表达,Pearson相关性分析宫颈癌组织中circDDX17和miR-224-3p表达的相关性。体外培养SiHa细胞,分为对照组、pcDNA组、pcDNA-circDDX17组、anti-miR-NC组、miR-224-3pInhibitor组和pcDNA-circDDX17+miR-224-3pmimic组,用X线照射,克隆形成实验检测SiHa细胞存活分数和克隆形成数,流式细胞术检测其细胞凋亡率。双荧光素酶报告基因实验验证circDDX17与miR-224-3p调控关系。结果与癌旁组织比较,宫颈癌组织中circDDX17表达降低(P<0.05),miR-224-3p表达升高(P<0.05),且宫颈癌组织中circDDX17与miR-224-3p的表达呈负相关(r=-0.981,P<0.05)。与对照组和pcDNA组(或anti-miR-NC组)比较,pcDNA-circDDX17组(或miR-224-3pInhibitor组)SiHa细胞经不同剂量(2、4、6、8Gy)X线照射后存活分数显著降低(P<0.05),经4GyX线照射后克隆数减少(P<0.05),而细胞凋亡率升高(P<0.05)。circDDX17可靶向结合并负调控miR-224-3p。与pcDNA-circDDX17组比较,pcDNA-circDDX17+miR-224-3pmimic组SiHa细胞经不同剂量(2、4、6、8Gy)X线照射后存活分数显著升高(P<0.05),经4GyX线照射后克隆数增加(P<0.05),而细胞凋亡率降低(P<0.05)。结论circDDX17可通过靶向抑制miR-224-3p的表达增强宫颈癌SiHa细胞的放射敏感性。

Objective To investigate the effect of circDDX17 on the radiosensitivity of cervical cancer cells SiHa and its possible mechanism. Methods Cervical cancer tissues and adjacent tissues were collected from 41 cervical cancer patients treated in the Second Hospital of Baoding from August 2018 to August 2020,and qRT-PCR was used to detect the expression of circDDX17 and miR-224-3 p in the tissues. Pearson correlation analysis was used to analyze the correlation between the expression of circDDX17 and miR-224-3 p in cervical cancer tissues. SiHa cells were cultured in vitro, and divided into control group, pcDNA group, pcDNA-circDDX17 group, anti-miR-NC group, miR-224-3 p Inhibitor group, and pcDNA-circDDX17+miR-224-3 p mimic group, and then they were irradiated with X-ray. Colony formation assay was used to detect cell survival fraction and colony formation number. Flow cytometry was used to detect cell apoptosis. The dual luciferase reporter gene experiment verified the regulatory relationship between circDDX17 and miR-224-3 p. Results Compared with the adjacent tissues, the expression of circDDX17 in cervical cancer tissues decreased(P<0.05), but the expression of miR-224-3 p increased(P<0.05). The expression of circDDX17 and miR-224-3 p in cervical cancer tissue was negatively correlated(r=-0.981, P<0.05). Compared with the control group and pcDNA group(or anti-miR-NC group), the survival score of SiHa cells in the pcDNA-circDDX17 group(or miR-224-3 p Inhibitor group) was significantly reduced after X-ray irradiation at different doses(2, 4, 6, 8 Gy)(P<0.05), and the number of colonies after 4 Gy X-ray irradiation was reduced(P<0.05), but the apoptotic rate was increased(P<0.05). circDDX17 could target and negatively regulate miR-224-3 p. Compared with the pcDNA-circDDX17 group, the survival score of SiHa cells in the pcDNA-circDDX17+miR-224-3 p mimic group was significantly increased after X-ray irradiation at different doses(2, 4, 6, 8 Gy)(P<0.05), and the number of colonies after 4 Gy X-ray irradiation was increased(P<0.05), but the apoptotic rate was reduced(P<0.05). Conclusions CircDDX17 can enhance the radiosensitivity of cervical cancer cells SiHa by targeting to inhibit the expression of miR-224-3 p.