摘要
目的:探讨环状RNA(circular RNA,circRNA)hsa_circ_0061137对宫颈癌(cervical cancer,CC)生长转移的影响及其相关作用机制。方法:取CC组织样本(93例)及癌旁组织样本(93例),正常宫颈上皮细胞系(End1/E6E7)和CC细胞系(HeLa、SiHa、C-33A、CaSki),用qRT-PCR法检测组织和细胞中hsa_circ_0061137、miR-217、ARL6IP1的表达;双荧光素酶报告实验验证hsa_circ_0061137、ARL6IP1与miR-217的靶向调控作用。敲低CC细胞系(HeLa、SiHa)中hsa_circ_0061137及HeLa细胞共转染si-hsa_circ_0061137和miR-217抑制物(miR-217 inhibitor)后,用CCK8法检测细胞增殖;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;Western blot检测上皮间质转化(epithelial-mesenchymal transition,EMT)相关蛋白(E-cadherin、Vimentin)的表达;qRT-PCR法检测各组细胞中hsa_circ_0061137、miR-217、ARL6IP1的表达。裸鼠荷瘤实验检测敲低hsa_circ_0061137后对肿瘤生长的影响。结果:hsa_circ_0061137、ARL6IP1在CC组织及细胞系中表达水平显著高于正常宫颈组织及正常宫颈上皮细胞(P<0.05);miR-217在CC组织及细胞系中表达水平显著低于正常宫颈组织及正常宫颈上皮细胞(P<0.05)。hsa_circ_0061137、ARL6IP1分别与miR-217之间存在靶向负调控关系。敲低hsa_circ_0061137,可抑制CC细胞增殖、迁移、侵袭及EMT特性,并促进CC细胞凋亡(P<0.05);miR-217 inhibitor可部分逆转hsa_circ_0061137敲低发挥的抗CC生长及转移作用(P<0.05)。裸鼠荷瘤实验证实hsa_circ_0061137敲低可显著减弱瘤体生长增殖,并抑制ARL6IP1表达(P<0.05)。结论:沉默hsa_circ_0061137可发挥抗CC增殖及转移作用,其作用可能与靶向miR-217/ARL6IP1轴有关。
Objective:To investigate the effect of circRNA hsa_circ_0061137 on the growth and metastasis of cervical cancer and its mechanism.Methods:CC tissue samples(93 cases),adjacent tissue samples(93 cases),normal cervical epithelial cell lines(End1/E6E7)and CC cell lines(HeLa,SiHa,C-33A,CaSki)were taken.qRT-PCR was used to detect the expression of hsa_circ_0061137,miR-217,ARL6IP1 in tissues and cells.Dual luciferase reporter test was used to verify the targeted regulation of hsa_circ_0061137,ARL6IP1 and miR-217.After knocking down hsa_circ_0061137 in CC cell lines(HeLa,SiHa)and co-transfecting with si-hsa_circ_0061137 and miR-217 inhibitor in HeLa cells.CCK8 method was used to detect cell proliferation.Flow cytometry was used to detect cell apoptosis.Transwell method was used to detect cell migration and invasion.Western blot was used to detect EMT-related proteins(E-cadherin,Vimentin).qRT-PCR method was used to detect the expression of hsa_circ_0061137,miR-217,ARL6IP1 in each group of cells.Tumor-bearing test in nude mice was used to detect the effect of knocking down hsa_circ_0061137 on tumor growth.Results:The expression levels of hsa_circ_0061137 and ARL6IP1 in CC tissues and cell lines were significantly higher than those in normal cervical tissues and normal cervical epithelial cells(P<0.05).The expression level of miR-217 in CC tissues and cell lines was significantly lower than that of normal cervical tissues and normal cervical epithelial cells(P<0.05).There was a targeted negative regulatory relationship between hsa_circ_0061137,ARL6IP1 and miR-217,respectively.Knockdown of hsa_circ_0061137 could inhibit CC cell proliferation,migration,invasion and EMT characteristics,and promote CC cell apoptosis(P<0.05).miR-217 inhibitor could partially reverse the anti-CC growth and metastasis effects of hsa_circ_0061137 knockdown(P<0.05).The nude mouse tumor-bearing test confirmed that hsa_circ_0061137 knockdown could significantly reduce tumor growth and proliferation,and inhibit the expression of ARL6IP1(P<0.05).Conclusion:Silencing hsa_circ_0061137 can play an anti-CC proliferation and metastasis role,and its action may be related to the target miR-217/ARL6IP1 axis.
作者
张小利
董丽
杜晨旭
崔玲玲
张适
谢林森
ZHANG Xiaoli;DONG Li;DU Chenxu;CUI Lingling;ZHANG Shi;XIE Linsen(Department of Clinical Laboratory,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Henan Zhengzhou 450001,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第3期435-442,共8页
Journal of Modern Oncology
基金
河南省高等学校重点科研项目(编号:21A310029)。
