Cai Yantao, Li Chuyu, Peng Fang, Yin Shuanghong, Liang Huiyi, Su Jiyan, Li Lin, Yang Anping, Liu Hui, Yang Chuansheng, Luo Dixian, Xia Chenglai
Affiliated Foshan Maternity and Chlid Healthcare Hospital, Southern Medical University, Foshan, China.
Department of Pharmacology, Guangdong Medical University, Zhanjiang, China.
Front Genet. 2021 Dec 17;12:779195. doi: 10.3389/fgene.2021.779195. eCollection 2021.
In recent years, circular RNAs (circRNAs) have been reported to serve as essential regulators in several human cancers. Nevertheless, the function and mechanism of circRNAs in cervical cancer remain elusive. Flow cytometry assays were performed to measure cell apoptosis and cell cycle. Colony Formation and transwell chamber were performed to measure cell migration and invasion. Double luciferase reporter for gene analysis was used to detect the interaction between hsa-circRNA_0001400, miR-326, and Akt. Relative protein levels were determined by immunoblotting and relative gene levels were determined by quantitative real-time PCR. Tumor Xenograft Modeling was used to evaluate the effect of hsa_circRNA_0001400_siRNA . In the present study, we showed that hsa_circRNA_0001400 was highly expressed in cervical cancer tissues relative to in matched normal tissue. We found that hsa_circRNA_0001400_siRNA significantly promoted the apoptosis of cervical cancer cells and arrested the cell cycle and migration of cervical cancer cells. We showed that hsa_circRNA_0001400_siRNA can inhibit the protein expression of Akt and that the inhibition of miR-326 could rescue the inhibition of Akt in cervical cancer cells. We found that has-miR-326 was downregulated in cervical cancer tissues and hsa_circRNA_0001400_siRNA could increase the gene expression of has-miR-326. We also observed that hsa_circRNA_0001400_siRNA inhibited the growth and angiogenesis of SiHa xenografts in nude mice. In conclusion, this study provides evidence that the hsa_circRNA_0001400-miR-326-Akt network promotes cervical cancer progression. Notably, our findings demonstrate the novel antitumor effects of hsa_circRNA_0001400_siRNA in cervical cancer.
近年来,环状RNA(circRNAs)已被报道在多种人类癌症中作为重要调节因子发挥作用。然而,circRNAs在宫颈癌中的功能和机制仍不清楚。进行流式细胞术检测以测量细胞凋亡和细胞周期。进行集落形成实验和Transwell小室实验以测量细胞迁移和侵袭。使用双荧光素酶报告基因分析来检测hsa-circRNA_0001400、miR-326和Akt之间的相互作用。通过免疫印迹法测定相对蛋白水平,通过定量实时PCR测定相对基因水平。采用肿瘤异种移植模型评估hsa_circRNA_0001400_siRNA的作用。在本研究中,我们发现相对于配对的正常组织,hsa_circRNA_0001400在宫颈癌组织中高表达。我们发现hsa_circRNA_0001400_siRNA显著促进宫颈癌细胞凋亡,并使宫颈癌细胞的细胞周期停滞以及抑制其迁移。我们发现hsa_circRNA_0001400_siRNA可抑制Akt的蛋白表达,并且抑制miR-326可挽救对宫颈癌细胞中Akt的抑制作用。我们发现has-miR-326在宫颈癌组织中表达下调,hsa_circRNA_0001400_siRNA可增加has-miR-326的基因表达。我们还观察到hsa_circRNA_0001400_siRNA抑制了裸鼠中SiHa异种移植瘤的生长和血管生成。总之,本研究提供了证据表明hsa_circRNA_0001400-miR-326-Akt网络促进宫颈癌进展。值得注意的是,我们的研究结果证明了hsa_circRNA_0001400_siRNA在宫颈癌中的新型抗肿瘤作用。