Research Paper Volume 13, Issue 6 pp 9119—9134

Silencing of circular RNA_0000326 inhibits cervical cancer cell proliferation, migration and invasion by boosting microRNA-338-3p-dependent down-regulation of CDK4

Zhaoxin Wang1,2, , Chenchen Ren1, &, , Li Yang1, , Xiaoan Zhang3, , Jiaxi Liu1, , Yuanhang Zhu1, , Dongyuan Jiang1, ,

  • 1 Department of Obstetrics and Gynecology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P. R. China
  • 2 Academy of Medical Science, Zhengzhou University, Zhengzhou 450052, P. R. China
  • 3 Department of Imaging, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, P. R. China

Received: March 18, 2020       Accepted: June 29, 2020       Published: March 17, 2021
How to Cite

Copyright: © 2021 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Cervical cancer is one of the leading causes of cancer-related death among women, which is attributed partly by limited treatment options. Recent studies have provided in-depth explanations regarding the role of circular RNA in cancers. We aimed to investigate the role of circular RNA_0000326 in cervical cancer. Bioinformatics analysis revealed a high circ_0000326 expression in cervical cancer. Cervical cancer cells and tissues were also observed to have elevated levels of circ_0000326 and the upregulation of circ_0000326 depended on the stage of cancer. Transfection with siRNA of circ_0000326 resulted in the inhibition of proliferation, migration and cell cycle of cancer cells. Interestingly, we confirmed that circ_0000326 served as a sponge for microRNA-338-3p and that the miRNA bound to Cyclin-dependent kinase 4. In the presence of microRNA-338-3p mimic or silencing of circ_0000326, Cyclin-dependent kinase 4 expression was decreased. Transfection with microRNA-338-3p mimic inhibited cell clone formation and proliferation. Moreover, in vivo experiment revealed that the injection of shRNA-circ_0000326 lentivirus suppressed tumor growth and decreased Cyclin-dependent kinase 4 expression. Taken altogether, our results showed that circ_0000326 exerted oncogenic effects on cervical cancer by upregulating Cyclin-dependent kinase 4 via sponging microRNA-338-3p. This systematic investigation on circ_0000326 could provide further insight into cervical cancer.


RIP: RNA immunoprecipitation; CDK4: Cyclin-dependent kinase 4; EdU: 5-ethynyl-2’-deoxyuridine; CCK8: cell counting kit-8; HPV: human papillomavirus; VEGF: vascular endothelial growth factor; circRNAs: circular RNAs; miRNA: microRNA; GEO: Gene Expression Omnibus; FDR: False positive discovery; DEGs: differentially expressed genes; FIGO: Federation of Gynecology and Obstetrics; ATCC: American Type Culture Collection; DMEM: Dulbecco's modified Eagle Medium; FBS: bovine serum; RT-qPCR: reverse transcription quantitative polymerase chain reaction; ABI: ABI7500 Real-Time PCR instrument; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MMP: matrix metallopeptidase; RLU: relative light unit; DAB: diaminobenzidine; DEPC: diethyl phosphorocyanidate; FISH: Fluorescence in situ hybridization; DAPI: 4'6-diamidino-2-phenylindole; CCK8: Cell counting kit-8; EdU: 5-ethynyl-2'-deoxyuridine; RIP: RNA immunoprecipitation.