1Department of Internal Medicine, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, China; 2Department of Thoracic Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, China; 3Department of Gynecology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, China; 4Department of Radiation Oncology, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, Shenyang 110042, China
Abstract：Objective To investigate the biological role of circ0004771 in lung adenocarcinoma. Methods Lung adenocarcinoma A549 cells were cultured and divided into the negative control (si-NC) and circ0004771 siRNA (si-circ0004771) groups. Si-NC and circ0004771 siRNA were transfected into A549 cells in the corresponding groups. After the transfection of the cells in the si-NC and si-circ0004771 groups, the expression levels of circ0004771 and miR-339-5p were detected via real-time fluorescent quantitative polymerase chain reaction; cell viability was determined through cell counting kit-8; cell proliferation was detected by using the EdU assay; and cell migration and invasion were detected through the Transwell chamber assay. Results In the A549 cells in the si-NC and si-circ0004771 groups, the relative expression levels of the circ0004771 gene were 3.07±0.07 and 0.68±0.04, respectively, and the relative expression levels of miR-339-5p were 1.14±0.13 and 2.33±0.07, respectively; these differences were statistically significant (t=51.34,13.96, all P values < 0.001). The absorbance values of the A549 cells in the si-NC and si-circ0004771 groups at 0, 24, and 48 h were 0.21±0.02, 0.35±0.05, and 0.65±0.04, respectively, and 0.21±0.01,0.33±0.04, and 0.59±0.05, respectively; no significant difference was observed (all P values > 0.05). However, at 72 h, the absorbance of the si-circ0004771 group was 0.92±0.15, which was less than that of the si-NC group (1.32±0.04); this difference was statistically significant (t=4.46, P=0.011). In the EdU experiment, the positive rate of cells in the si-circ0004771 group was 47.97%±4.68%, which was lower than the positive rate of 58.61%±2.15% shown by the si-NC group; this difference was statistically significant (t=3.58, P=0.023). After transfection, the cell migration and invasion rates in the si-circ0004771 group were 52.27%±2.92% and 55.33%±6.29%, respectively, and were all lower than the cell migration and invasion rates of 99.79%±4.23% and 98.67% ±5.72% in the si-NC group; this difference was statistically significant (t=16.26, 8.83, all P values < 0.001). Conclusions Down regulating the expression level of circ0004771 in lung adenocarcinoma cell line A549 can reduce the viability and proliferation of A549 cells and inhibit the migration and invasion of A549 cells; the mechanism of this effect may be related to the upregulation of mir-339-5p expression.
杨露, 丁剑桥, 陈金鑫, 张叶, 张亮, 项博文, 秦宝丽. 环状RNA circ0004771对肺腺癌细胞A549的增殖、迁移与侵袭的影响[J]. 中华解剖与临床杂志, 2022, 27(1): 42-47.
Yang Lu, Ding Jianqiao, Chen Jinxin, Zhang Ye, Zhang Liang, Xiang Bowen, Qin Baoli. Effects of circRNA circ0004771 on the proliferation, migration, and invasion of lung adenocarcinoma cell line A549. Chinese Journal of Anatomy and Clinics, 2022, 27(1): 42-47.
Siegel RL, Miller KD, Jemal A.Cancer statistics, 2019[J]. CA Cancer J Clin, 2019,69(1):7-34. DOI: 10.3322/caac.21551.
[2]
Bray F, Ferlay J, Soerjomataram I, et al.Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018,68(6):394-424. DOI: 10.3322/caac.21492.
[3]
Zhou J, Xiao H, Yang X, et al.Long noncoding RNA CASC9.5 promotes the proliferation and metastasis of lung adenocarcinoma[J]. Sci Rep, 2018,8(1):37. DOI: 10.1038/s41598-017-18280-3.
[4]
Cancer Genome Atlas Research Network. Comprehensive molecular profiling of lung adenocarcinoma[J]. Nature, 2014, 511(7511): 543-550. DOI: 10.1038/nature13385.
[5]
Gao LM, Xu SF, Zheng Y, et al.Long non-coding RNA H19 is responsible for the progression of lung adenocarcinoma by mediating methylation-dependent repression of CDH1 promoter[J]. J Cell Mol Med, 2019,23(9):6411-6428. DOI: 10.1111/jcmm.14533.
[6]
Gettinger S, Lynch T.A decade of advances in treatment for advanced non-small cell lung cancer[J]. Clin Chest Med, 2011, 32(4): 839-851. DOI: 10.1016/j.ccm.2011.08.017.
[7]
Capel B, Swain A, Nicolis S, et al.Circular transcripts of the testis-determining gene Sry in adult mouse testis[J]. Cell, 1993,73(5):1019-1030. DOI: 10.1016/0092-8674(93)90279-y.
Chen I, Chen CY, Chuang TJ.Biogenesis, identification, and function of exonic circular RNAs[J]. Wiley Interdiscip Rev RNA, 2015,6(5):563-579. DOI: 10.1002/wrna.1294.
[10]
Jeck WR, Sorrentino JA, Wang K, et al.Circular RNAs are abundant, conserved, and associated with ALU repeats[J]. RNA, 2013,19(2):141-157. DOI: 10.1261/rna.035667.112.
[11]
Wang X, Wang R, Wu Z, et al.Circular RNA ITCH suppressed prostate cancer progression by increasing HOXB13 expression via spongy miR-17-5p[J]. Cancer Cell Int, 2019, 19: 328. DOI: 10.1186/s12935-019-0994-8.
[12]
Sui MH, Zhang WW, Geng DM, et al.CircPRKCI regulates proliferation, migration and cycle of lung adenocarcinoma cells by targeting miR-219a-5p-regulated CAMK1D[J]. Eur Rev Med Pharmacol Sci, 2021,25(4):1899-1909. DOI: 10.26355/eurrev_202102_25085.
[13]
Yao Y, Hua Q, Zhou Y.CircRNA has_circ_0006427 suppresses the progression of lung adenocarcinoma by regulating miR-6783-3p/DKK1 axis and inactivating Wnt/β-catenin signaling pathway[J]. Biochem Biophys Res Commun, 2019, 508(1): 37-45. DOI: 10.1016/j.bbrc.2018.11.079.
[14]
Liu Y, Jiang Y, Xu L, et al.circ-NRIP1 promotes glycolysis and tumor progression by regulating miR-186-5p/MYH9 axis in gastric cancer[J]. Cancer Manag Res, 2020,12:5945-5956. DOI: 10.2147/CMAR.S245941.
[15]
Li X, Ma N, Zhang Y, et al.Circular RNA circNRIP1 promotes migration and invasion in cervical cancer by sponging miR-629-3p and regulating the PTP4A1/ERK1/2 pathway[J]. Cell Death Dis, 2020, 11(5): 399. DOI: 10.1038/s41419-020-2607-9.
[16]
Li M, Cai J, Han X, et al.Downregulation of circNRIP1 suppresses the paclitaxel resistance of ovarian cancer via regulating the miR-211-5p/HOXC8 axis[J]. Cancer Manag Res, 2020, 12: 9159-9171. DOI: 10.2147/CMAR.S268872.
[17]
Chen F, Huang C, Wu Q, et al.Circular RNAs expression profiles in plasma exosomes from early-stage lung adenocarcinoma and the potential biomarkers[J]. J Cell Biochem, 2020,121(3):2525-2533. DOI: 10.1002/jcb. 29475.
[18]
Vora N, Reckamp KL.Non-small cell lung cancer in the elderly: defining treatment options[J]. Semin Oncol, 2008,35(6): 590-596. DOI: 10.1053/j.seminoncol.2008.08.009.
[19]
Bade BC, Dela Cruz CS.Lung cancer 2020: epidemiology, etiology, and prevention[J]. Clin Chest Med, 2020,41(1):1-24. DOI: 10.1016/j.ccm.2019.10.001.
Lin YW, Lee B, Liu PS, et al.Receptor-Interacting protein 140 orchestrates the dynamics of macrophage M1/M2 polarization[J]. J Innate Immun, 2016, 8(1): 97-107. DOI: 10.1159/000433539.
[22]
Zhou S, Guo Z, Zhou C, et al.circ_NRIP1 is oncogenic in malignant development of esophageal squamous cell carcinoma (ESCC) via miR-595/SEMA4D axis and PI3K/AKT pathway[J]. Cancer Cell Int, 2021, 21(1): 250. DOI: 10.1186/s12935-021-01907-x.
[23]
Xie R, Tang J, Zhu X, et al. Silencing of hsa_circ_0004771 inhibits proliferation and induces apoptosis in breast cancer through activation of miR-653 by targeting ZEB2 signaling pathway[J]. Biosci Rep, 2019, 39(5): BSR20181919. DOI: 10.1042/BSR20181919.
[24]
Pan B, Qin J, Liu X, et al.Identification of serum exosomal hsa-circ-0004771 as a novel diagnostic biomarker of colorectal cancer[J]. Front Genet, 2019,10:1096. DOI: 10.3389/fgene.2019.01096.
[25]
Li T, Shao Y, Fu L, et al.Plasma circular RNA profiling of patients with gastric cancer and their droplet digital RT-PCR detection[J]. J Mol Med (Berl), 2018,96(1):85-96. DOI: 10.1007/s00109-017-1600-y.
[26]
Bushati N, Cohen SM. microRNA functions[J]. Annu Rev Cell Dev Biol, 2007,23:175-205. DOI: 10.1146/annurev.cellbio.23.090506.123406.
[27]
Wang YL, Chen CM, Wang XM, et al.Effects of miR-339-5p on invasion and prognosis of hepatocellular carcinoma[J]. Clin Res Hepatol Gastroenterol, 2016,40(1):51-56. DOI: 10.1016/j.clinre.2015.05.022.
[28]
Wu ZS, Wu Q, Wang CQ, et al.MiR-339-5p inhibits breast cancer cell migration and invasion in vitro and may be a potential biomarker for breast cancer prognosis[J]. BMC Cancer, 2010, 10: 542. DOI: 10.1186/1471-2407-10-542.
[29]
Li P, Liu H, Li Y, et al.miR-339-5p inhibits lung adenocarcinoma invasion and migration by directly targeting BCL6[J]. Oncol Lett, 2018, 16(5):5785-5790. DOI: 10.3892/ol.2018.9376.
[30]
Huang E, Fu J, Yu Q, et al.CircRNA hsa_circ_0004771 promotes esophageal squamous cell cancer progression via miR-339-5p/CDC25A axis[J]. Epigenomics, 2020,12(7):587-603. DOI: 10.2217/epi-2019-0404.
2022, Vol. 27 
