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circ-PRKDC通过调控miR-505-3p影响肺癌细胞增殖、凋亡和放射敏感性机制研究
袁小笋1, 张蕾1, 饶石磊2, 张凯2, 马慧利1, 李长生1, 张敬伟1, 任中海1
1南阳市中心医院肿瘤内科二病区 473000; 2南阳市中心医院放疗科 473000
circ-PRKDC affects the proliferation, apoptosis and radiosensitivity of lung cancer cells by regulating miR-505-3p
Yuan Xiaosun1, Zhang Lei1, Rao Shilei2, Zhang Kai2, Ma Huili1, Li Changsheng1, Zhang Jingwei1, Ren Zhonghai1
1Second Ward of Department of Oncology, Nanyang Central Hospital, Nanyang 473000, China; 2Department of Radiotherapy, Nanyang Central Hospital, Nanyang 473000, China
Abstract：Objective To investigate the effect of circ-PRKDC on lung cancer cell proliferation, apoptosis and radiosensitivity and its molecular mechanism. Methods Normal lung epithelial cells BEAS-2B and lung cancer cell lines NCI-H1299, NCI-H2170, NCI-H1975 were cultured. NCI-H1299 cells were divided into the si-NC, si-PRKDC, pcDNA-NC, pcDNA-PRKDC, miR-NC, miR-505-3p, anti-miR-NC, anti-miR-505-3p, si-PRKDC+anti-miR-NC and si-PRKDC+anti-miR-505-3p groups. RT-qPCR was used to detect the expression levels of circ-PRKDC and miR-505-3p. Western blot was employed to measure the protein expression. MTT was used to detect cell proliferation. Flow cytometry was utilized to detect cell apoptosis. Plate clone formation assay was conducted to detect the cell radiosensitivity. Dual luciferase reporter assay was performed to analyze the targeting relationship between circ-PRKDC and miR-505-3p. Results Compared with normal lung epithelial cells BEAS-2B, the expression levels of circ-PRKDC in the lung cancer cell lines NCI-H1299, NCI-H2170 and NCI-H1975 were significantly up-regulated (3.65, 3.10, 2.67 vs. 1.00, all P<0.05), whereas those of miR-505-3p were significantly down-regulated (0.42, 0.50, 0.54 vs. 1.02, all P<0.05). After low expression of circ-PRKDC, the expression level of CyclinD1 was significantly down-regulated (0.42 vs. 0.81, P<0.05), whereas those of Cleaved-caspase-3(0.71 vs. 0.33, P<0.05) and γ-H2AX (0.89 vs. 0.46, P<0.05) were significantly up-regulated, the cell A value was significantly decreased (0.413 vs. 0.839, P<0.05), cell apoptosis rate was significantly increased (20.35 vs. 6.21, P<0.05), cell survival fraction was significantly decreased (P<0.05), and β-catenin expression was significantly down-regulated (0.35 vs. 0.73, P<0.05). After high expression of miR-505-3p, the expression level of CyclinD1 was significantly down-regulated (0.34 vs. 0.83, P<0.05), those of Cleaved-caspase-3(0.65 vs. 0.32, P<0.05) and γ-H2AX (0.96 vs. 0.45, P<0.05) were significantly up-regulated, the cell A value was significantly decreased (0.386 vs. 0.851, P<0.05), the apoptosis rate was significantly increased (16.38 vs. 6.20, P<0.05), and the cell survival fraction was significantly decreased (P<0.05).compared with miR-NC, the luciferase activity of miR-505-3p group transfected with circ-PRKDC wild-type reporter plasmid was significantly decreased (0.44 vs. 1.00, P<0.05). Down-regulation of miR-505-3p could reverse the effect of low expression of circ-PRKDC on the proliferation, apoptosis, radiosensitivity and β-catenin expression of NCI-H1299 cells. Conclusion Low expression of circ-PRKDC may inhibit lung cancer cell proliferation, promote cell apoptosis and enhance cell radiosensitivity by up-regulating miR-505-3p, which is probably associated with the Wnt/β-catenin signaling pathway.
Yuan Xiaosun,Zhang Lei,Rao Shilei et al. circ-PRKDC affects the proliferation, apoptosis and radiosensitivity of lung cancer cells by regulating miR-505-3p[J]. Chinese Journal of Radiation Oncology, 2021, 30(12): 1309-1315.
[1] 朱鲁程,许雅思,夏冰,等. 肺癌放射敏感性标志物研究进展[J]. 中华放射肿瘤学杂志,2018, 27(4):420-424. DOI:10.3760/cma.j.issn.1004-4221.2018.04.018. Zhu LC, Xu YS, Xia B, et al. Research progress on biomarkers in predicting radiosensitivity of non-small cell lung cancer[J]. Chin J Radiat Oncol, 2018, 27(4):420-424. DOI:10.3760/cma.j.issn.1004-4221.2018.04.018. [2] 李光烈,阎英. 非小细胞肺癌放射敏感性标志物[J]. 国际肿瘤学杂志,2020, 47(4):236-239. DOI:10.3760/cma.j.cn371439-20190923-00009. Li GL, Yan Y. Radiosensitive markers of non-small cell lung cancer[J]. J Int Oncol, 2020, 47(4):236-239. DOI:10.3760/cma.j.cn371439-20190923-00009. [3] Zhang H, Shen Y, Li Z, et al. The biogenesis and biological functions of circular RNAs and their molecular diagnostic values in cancers[J]. J Clin Lab Anal, 2020, 34(1):e23049. DOI:10.1002/jcla.23049. [4] Chen H, Pei L, Xie P, et al. Circ-PRKDC contributes to 5-fluorouracil resistance of colorectal cancer cells by regulating miR-375/FOXM1 axis and Wnt/β-catenin pathway[J]. Onco Targets Ther, 2020, 13(1):5939-5953. DOI:10.2147/OTT. S253468. [5] Jin C, Wang A, Liu L, et al. Hsa_circ_0136666 promotes the proliferation and invasion of colorectal cancer through miR-136/SH2B1 axis[J]. J Cell Physiol, 2019, 234(5):7247-7256. DOI:10.1002/jcp.27482. [6] Liu LH, Tian QQ, Liu J, et al. Upregulation of hsa_circ_0136666 contributes to breast cancer progression by sponging miR-1299 and targeting CDK6[J]. J Cell Biochem, 2019, 120(8):12684-12693. DOI:10.1002/jcb.28536. [7] 宋秀军,江其生,高俊,等.miR-505、miR-505*在非小细胞肺癌患者组织及血清中的表达及意义[J]. 现代肿瘤医学,2017, 25(24):3978-3981. DOI:10.3969/j.issn.1672-4992.2017.24.014. Song XJ, Jiang QS, Gao J, et al. The expression of miR-505, miR-505* in FFPE and serum of NSCLC patients[J]. J Mod Oncol, 2017, 25(24):3978-3981. DOI:10.3969/j.issn.1672-4992.2017.24.014. [8] Cheng Z, Wang B, Zhang C. MicroRNA-505-3p inhibits development of glioma by targeting HMGB1 and regulating AKT expression[J]. Oncol Lett, 2020, 20(2):1663-1670. DOI:10.3892/ol.2020.11714. [9] Zhao P, Guan H, Dai Z, et al. Long noncoding RNA DLX6-AS1 promotes breast cancer progression via miR-505-3p/RUNX2 axis[J]. Eur J Pharmacol, 2019, 865(1):172778. DOI:10.1016/j.ejphar.2019.172778. [10] 王玉佩. 线粒体F1Fo ATP酶抑制因子调节非小细胞肺癌辐射敏感性的机制研究[D]. 北京:中国科学院大学(中国科学院近代物理研究所),2019. Wang YP. Study on the mechanism of mitochondrial F1Fo ATPase inhibitor in regulating radiosensitivity of non-small cell lung cancer[D]. Beijing:University of Chinese Academy of Sciences (Institute of Modern Physics, Chinese Academy of Sciences), 2019. [11] Ma Y, Zhang X, Wang YZ, et al. Research progress of circular RNAs in lung cancer[J]. Cancer Biol Ther, 2019, 20(2):123-129. DOI:10.1080/15384047.2018.1523848. [12] Zhang C, Zhou H, Yuan K, et al. Overexpression of hsa_circ_0136666 predicts poor prognosis and initiates osteosarcoma tumorigenesis through miR-593-3p/ZEB2 pathway[J]. Aging, 2020, 12(11):10488-10496. DOI:10.18632/aging.103273. [13] 韩宝林,王轩,李立芳,等.γH2AX对肺癌放射敏感性的预测效果[J]. 天津医药,2014, 42(9):867-869. DOI:10.3969/j.issn.0253-9896.2014.09.005. Han BL, Wang X, Li LF, et al. Assessing radiosensitivity of lung cancer with the expression of γH2AX[J]. Tianjin Med J, 2014, 42(9):867-869. DOI:10.3969/j.issn.0253-9896.2014.09.005. [14] Tang Y, Wu B, Huang S, et al. Downregulation of miR-505-3p predicts poor bone metastasis-free survival in prostate cancer[J]. Oncol Rep, 2019, 41(1):57-66. DOI:10.3892/or.2018.6826. [15] Yang K, Tong L, Li K, et al. A SRSF1self-binding mechanism restrains Mir505-3p from inhibiting proliferation of neural tumor cell lines[J]. Anticancer Drugs, 2018, 29(1):40-49. DOI:10.1097/CAD.0000000000000564. [16] Wei G, Lu T, Shen J, et al. LncRNA ZEB1-AS1 promotes pancreatic cancer progression by regulating miR-505-3p/TRIB2 axis[J]. Biochem Biophys Res Commun, 2020, 528(4):644-649. DOI:10.1016/j.bbrc.2020.05.105. [17] Yang Y, Zhou H, Zhang G, et al. Targeting the canonical Wnt/β-catenin pathway in cancer radioresistance:Updates on the molecular mechanisms[J]. J Cancer Res Ther, 2019, 15(2):272-277. DOI:10.4103/jcrt. JCRT_421_18. [18] He H, Lin K, Su Y, et al. Overexpression of β-Catenin decreases the radiosensitivity of human nasopharyngeal carcinoma CNE-2 cells[J]. Cell Physiol Biochem, 2018,50(5):1929-1944. DOI:10.1159/000494873. [19] Han PB, Ji XJ, Zhang M, et al. Upregulation of lncRNA LINC00473 promotes radioresistance of HNSCC cells through activating Wnt/β-catenin signaling pathway[J]. Eur Rev Med Pharmacol Sci, 2018, 22(21):7305-7313. DOI:10.26355/eurrev_201811_16267. [20] Wu D, Li L, Yan W. Knockdown of TC-1 enhances radiosensitivity of non-small cell lung cancer via the Wnt/β-catenin pathway[J]. Biol Open, 2016,5(4):492-498. DOI:10.1242/bio.017608.
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