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Abstract: BackgroundCircular RNAs (circRNAs) are covalently closed single-stranded RNAs with multiple biological functions. circRNA.0007127 is derived from the carbon catabolite repression 4-negative on TATA-less (CCR4-NOT) complex subunit 2 (CNOT2), which was found to regulate tumor cell apoptosis through caspase pathway.
MethodsPotential circRNA.0007127 target microRNAs (miRNAs) were analyzed by miRanda, TargetScan, and RNAhybrid software, and the miRNAs with binding sites for apoptosis-related genes were screened. The roles of circRNA.0007127 and its downstream target, microRNA (miR)‍-513a-5p, were validated by quantitative real-time polymerase chain reaction (qPCR), flow cytometry, mitochondrial membrane potential, immunofluorescence, western blot, and caspase-8 (CASP8) protein activity in vitro in H₂O₂-induced k-562 cells. The circRNA.‍0007127‍‒‍miR-513a-5p and CASP8‍‒‍miR-513a-5p interactions were verified by luciferase reporter assays.
ResultsSilencing circRNA.0007127 decreased cell apoptosis by inhibiting CASP8 pathway activation in k-562 cells. Compared with the control group, the expression of CASP8 was reduced by 50% and the 43-kD fragment of CASP8 protein was significantly reduced (P≤0.05). The luciferase reporting assay showed that circRNA.0007127 combined with miR-513a-5p or CASP8, with extremely significant differences (P≤0.001). The overexpression of miR-513a-5p inhibited the gene expression level of CASP8 in a human myeloid leukemia cell model (75% change) and the level of a 43-kD fragment of CASP8 protein (P≤0.01). The rescue experiment showed that cotransfection with circRNA.0007127 small-interfering RNA (siRNA) and the miR-513a-5p inhibitor increased CASP8 gene expression and the apoptosis rate, suggesting that the miR-513a-5p inhibitor is a circRNA.0007127 siRNA antagonist.
ConclusionscircRNA.0007127 regulates K-562 cell apoptosis through the miR-513a-5p/CASP8 axis, which can serve as a novel powerful molecular target for k-562 cells.

CircRNA.0007127通过miR-513a-5p/CASP8轴触发K-562细胞凋亡

[1]BenetatosL, BenetatouA, VartholomatosG, 2020. Long non-coding RNAs and MYC association in hematological malignancies. Ann Hematol, 99(10):2231-2242.

[2]DaiN, QingY, CunYP, et al., 2018. miR-513a-5p regulates radiosensitivity of osteosarcoma by targeting human apurinic/apyrimidinic endonuclease. Oncotarget, 9(39):25414-25426.

[3]de AchaOP, RossiM, GorospeM, 2020. Circular RNAs in blood malignancies. Front Mol Biosci, 7:109.

[4]de BiasiS, GibelliniL, CossarizzaA, 2015. Uncompensated polychromatic analysis of mitochondrial membrane potential using JC-1 and multilaser excitation. Curr Protoc Cytom, 72:1-11.

[5]DroinN, GuéryL, BenikhlefN, et al., 2013. Targeting apoptosis proteins in hematological malignancies. Cancer Lett, 332(2):325-334.

[6]DuJ, ZhangLY, MaHZ, et al., 2020. Lidocaine suppresses cell proliferation and aerobic glycolysis by regulating circHOMER1/miR-138-5p/HEY1 axis in colorectal cancer. Cancer Manag Res, 12:5009-5022.

[7]DubeyM, NagarkotiS, AwasthiD, et al., 2016. Nitric oxide-mediated apoptosis of neutrophils through caspase-8 and caspase-3-dependent mechanism. Cell Death Dis, 7(9):e2348.

[8]GaoSJ, YuYY, LiuL, et al., 2019. Circular RNA hsa_circ_0007059 restrains proliferation and epithelial-mesenchymal transition in lung cancer cells via inhibiting microRNA-378. Life Sci, 233:116692.

[9]GriloAL, MantalarisA, 2019. Apoptosis: a mammalian cell bioprocessing perspective. Biotechnol Adv, 37(3):459-475.

[10]GuarnerioJ, BezziM, JeongJC, et al., 2016. Oncogenic role of fusion-circRNAs derived from cancer-associated chromosomal translocations. Cell, 165(2):289-302.

[11]HaoZ, HuS, LiuZ, et al., 2019. Circular RNAs: functions and prospects in glioma. J Mol Neurosci, 67(1):72-81.

[12]HuangY, ZhangP, DuL, et al., 2018. Mechanisms of recombinant adenovirus-mediated SD-HA fusion protein proliferation inhibition and induced apoptosis of K562 cells. Chin J Hematol, 39(4):314-319 (in Chinese).

[13]HungYC, PanTL, HuWL, 2016. Roles of reactive oxygen species in anticancer therapy with Salvia miltiorrhiza Bunge. Oxid Med Cell Longev, 2016:5293284.

[14]ItoK, InoueT, YokoyamaK, et al., 2011. CNOT2 depletion disrupts and inhibits the CCR4-NOT deadenylase complex and induces apoptotic cell death. Genes Cells, 16(4):368-379.

[15]JamalM, SongTB, ChenB, et al., 2019. Recent progress on circular RNA research in acute myeloid leukemia. Front Oncol, 9:1108.

[16]JungJH, ParkJE, SimDY, et al., 2019. Farnesiferol C induces apoptosis in chronic myelogenous leukemia cells as an imatinib sensitizer via caspase activation and HDAC (histone deacetylase) inactivation. Int J Mol Sci, 20(22):5535.

[17]JungJH, LeeHJ, KimJH, et al., 2020. Colocalization of MID1IP1 and c-Myc is critically involved in liver cancer growth via regulation of ribosomal protein L5 and L11 and CNOT2. Cells, 9(4):985.

[18]KellerN, OzmadenciD, IchimG, et al., 2018. Caspase-8 function, and phosphorylation, in cell migration. Semin Cell Dev Biol, 82:105-117.

[19]KerrJFR, WyllieAH, CurrieAR, 1972. Apoptosis: a basic biological phenomenon with wideranging implications in tissue kinetics. Br J Cancer, 26(4):239-257.

[20]KristensenLS, AndersenMS, StagstedLVW, et al., 2019. The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet, 20(11):675-691.

[21]LeeJ, JungJH, HwangJ, et al., 2019. CNOT2 is critically involved in atorvastatin induced apoptotic and autophagic cell death in non-small cell lung cancers. Cancers (Basel), 11(10):1470.

[22]LitwińskaZ, MachalińskiB, 2017. miRNAs in chronic myeloid leukemia: small molecules, essential function. Leuk Lymphoma, 58(6):1297-1305.

[23]LuMD, LiuD, LiYX, 2020. LINC01436 promotes the progression of gastric cancer via regulating miR-513a-5p/APE1 axis. Onco Targets Ther, 13:10607-10619.

[24]MandalR, BarrónJC, KostovaI, et al., 2020. Caspase-8: the double-edged sword. Biochim Biophys Acta Rev Cancer, 1873(2):188357.

[25]MarcondesNA, TerraSR, LastaCS, et al., 2019. Comparison of JC-1 and mitotracker probes for mitochondrial viability assessment in stored canine platelet concentrates: a flow cytometry study. Cytometry A, 95(2):214-218.

[26]MatsumotoT, JimiS, MigitaK, et al., 2019. FF-10501 induces caspase-8-mediated apoptotic and endoplasmic reticulum stress-mediated necrotic cell death in hematological malignant cells. Int J Hematol, 110(5):606-617.

[27]MeiM, WangYJ, LiZM, et al., 2019. Role of circular RNA in hematological malignancies (Review). Oncol Lett, 18(5):4385-4392.

[28]MemczakS, JensM, ElefsiniotiA, et al., 2013. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature, 495(7441):333-338.

[29]MutiP, DonzelliS, SacconiA, et al., 2018. MiRNA-513a-5p inhibits progesterone receptor expression and constitutes a risk factor for breast cancer: the hOrmone and Diet in the ETiology of breast cancer prospective study. Carcinogenesis, 39(2):98-108.

[30]ObengE, 2021. Apoptosis (programmed cell death) and its signals‒a review. Braz J Biol, 81(4):1133-1143.

[31]PanniS, LoveringRC, PorrasP, et al., 2020. Non-coding RNA regulatory networks. Biochim Biophys Acta Gene Regul Mech, 1863(6):194417.

[32]PerelmanA, WachtelC, CohenM, et al., 2012. JC-1: alternative excitation wavelengths facilitate mitochondrial membrane potential cytometry. Cell Death Dis, 3(11):e430.

[33]Rodriguez-GilA, RitterO, HornungJ, et al., 2016. HIPK family kinases bind and regulate the function of the CCR4-NOT complex. Mol Biol Cell, 27(12):1969-1980.

[34]SalzmanJ, ChenRE, OlsenMN, et al., 2013. Cell-type specific features of circular RNA expression. PLoS Genet, 9(9):e1003777.

[35]SlackFJ, ChinnaiyanAM, 2019. The role of non-coding RNAs in oncology. Cell, 179(5):1033-1055.

[36]SongS, LeeJY, ErmolenkoL, et al., 2020. Tetrahydrobenzimidazole TMQ0153 triggers apoptosis, autophagy and necroptosis crosstalk in chronic myeloid leukemia. Cell Death Dis, 11(2):109.

[37]ŠrámekJ, Němcová-FürstováV, KovářJ, 2021. Molecular mechanisms of apoptosis induction and its regulation by fatty acids in pancreatic β‍-cells. Int J Mol Sci, 22(8):4285.

[38]SuW, SunS, WangF, et al., 2019. Circular RNA hsa_circ_0055538 regulates the malignant biological behavior of oral squamous cell carcinoma through the p53/Bcl-2/caspase signaling pathway. J Transl Med, 17:76.

[39]TaylorRC, CullenSP, MartinSJ, 2008. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol, 9(3):231-241.

[40]TummersB, GreenDR, 2017. Caspase-8: regulating life and death. Immunol Rev, 277(1):76-89.

[41]WenN, LvQ, DuZG, 2020. MicroRNAs involved in drug resistance of breast cancer by regulating autophagy. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(9):690-702.

[42]XiaoY, MingX, WuJY, 2021. Hsa_circ_0002483 regulates miR-758-3p/MYC axis to promote acute myeloid leukemia progression. Hematol Oncol, 39(2):243-253.

[43]XuJ, WuKJ, JiaQJ, et al., 2020. Roles of miRNA and lncRNA in triple-negative breast cancer. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(9):673-689.

[44]YuH, LiSB, 2020. Role of LINC00152 in non-small cell lung cancer. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(3):179-191.

[45]YuanWB, ZhouR, WangJZ, et al., 2019. Circular RNA Cdr1as sensitizes bladder cancer to cisplatin by upregulating APAF1 expression through miR-1270 inhibition. Mol Oncol, 13(7):1559-1576.

[46]ZhuRY, QiXY, LiuCP, et al., 2020. The silent information regulator 1 pathway attenuates ROS-induced oxidative stress in Alzheimer’s disease. J Integr Neurosci, 19(2):321-332.