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山东大学学报 (医学版) ›› 2023, Vol. 61 ›› Issue (9): 38-46.doi: 10.6040/j.issn.1671-7554.0.2023.0140

• 基础医学 • 上一篇    下一篇

CircFAT1调节miR-296-3p/MAPRE1轴对鼻咽癌细胞增殖、凋亡和放疗敏感性的影响

曹华琳1,贾彦召2,曲莉1,尹昕1   

  1. 南阳市中心医院 1.耳鼻喉头颈外科;2.放疗科, 河南 南阳 473000
  • 收稿日期:2023-02-15 发布日期:2023-10-10
  • 通讯作者: 尹昕. E-mail:47795802@qq.com
  • 基金资助:
    河南省医学科技攻关项目(2020170394)

Impacts of CircFAT1 on the proliferation, apoptosis and radiosensitivity of nasopharyngeal carcinoma cells by regulating miR-296-3p/MAPRE1 axis

CAO Hualin1, JIA Yanzhao2, QU Li1, YIN Xin1   

  1. 1. Department of Otolaryngology, Head and Neck Surgery;
    2. Radiotherapy Department, Nanyang Central Hospital, Nanyang 473000, Henan, China
  • Received:2023-02-15 Published:2023-10-10

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摘要： 目的 研究环状RNA脂肪非典型钙黏蛋白1(CircFAT1)调节miR-296-3p/微管关联蛋白RP/EB家族成员1(MAPRE1)轴对鼻咽癌细胞增殖、凋亡和放疗敏感性的影响。 方法 以实时荧光定量PCR和免疫印迹法检测鼻咽上皮细胞NP69、人鼻咽癌细胞株CNE2及其放射线抵抗细胞CNE2-RR中CircFAT1、miR-296-3p、MAPRE1表达。将体外培养的CNE2细胞随机分为对照组、CircFAT1敲低组、阴性对照组、CircFAT1敲低+miR-296-3p inhibitor组,分组转染后以实时荧光定量PCR和免疫印迹法检测各组CNE2细胞CircFAT1、miR-296-3p、MAPRE1表达;以CCK-8法、5-乙炔基-2-脱氧尿苷(Edu)染色与流式细胞实验检测各组CNE2细胞增殖、凋亡。将体外培养的CNE2-RR细胞随机分为对照组、放射组、放射+阴性对照组、放射+CircFAT1敲低组、放射+CircFAT1敲低+miR-296-3p inhibitor组,分组转染后以实时荧光定量PCR和免疫印迹法检测各组CNE2-RR细胞CircFAT1、miR-296-3p、MAPRE1表达;以CCK-8法、Edu染色与流式细胞实验检测各组CNE2-RR细胞增殖、凋亡。以双荧光素酶报告基因实验检测CNE2-RR中CircFAT1对miR-296-3p的靶向调节与miR-296-3p对MAPRE1的靶向调节。 结果 与NP69细胞相比,CNE2、CNE2-RR细胞CircFAT1表达、MAPRE1 mRNA与蛋白表达升高(P<0.05),miR-296-3p表达降低(P<0.05);与CNE2细胞相比,CNE2-RR细胞CircFAT1表达、MAPRE1 mRNA与蛋白表达升高(P<0.05),miR-296-3p表达降低(P<0.05)。与对照组相比,CircFAT1敲低组CNE2细胞CircFAT1表达、MAPRE1 mRNA与蛋白表达、存活率、增殖率降低(P<0.05),miR-296-3p表达、凋亡率升高(P<0.05);阴性对照组CNE2细胞各指标无明显变化(P>0.05),miR-296-3p inhibitor可逆转敲低CircFAT1对CNE2细胞各指标的影响。与对照组、放射组分别相比,放射+CircFAT1敲低组CNE2-RR细胞CircFAT1表达、MAPRE1 mRNA与蛋白表达、存活率、增殖率降低(P<0.05),miR-296-3p表达、凋亡率升高(P<0.05);放射组、放射+阴性对照组CNE2-RR细胞各指标无明显变化(P>0.05),miR-296-3p inhibitor可逆转敲低CircFAT1对放射处理下CNE2-RR细胞各指标的影响。CircFAT1可靶向下调CNE2-RR细胞miR-296-3p表达,而miR-296-3p可靶向下调CNE2-RR细胞MAPRE1表达。 结论 敲低CircFAT1可通过上调miR-296-3p而降低MAPRE1表达,从而抑制鼻咽癌细胞增殖,促进其凋亡并增强其放射敏感性。

关键词: 环状RNA脂肪非典型钙黏蛋白1, 微管关联蛋白, 鼻咽癌, 增殖, 凋亡, 放疗敏感性

Abstract: Objective To explore the impacts of circular RNA fat atypical cadherin 1(CircFAT1)on the proliferation, apoptosis and radiosensitivity of nasopharyngeal carcinoma cells by regulating miR-296-3p/microtubule-associated protein RP/EB family member 1(MAPRE1)axis. Methods The expressions of CircFAT1, miR-296-3p and MAPRE1 in NP69, CNE2 and CNE2-RR cells were detected with real-time fluorescent quantitative PCR and Western blotting. CNE2 cells cultured in vitro were randomly divided into the control group, CircFAT1 knockdown group, negative control group, and CircFAT1 knockdown+miR-296-3p inhibitor group. After transfection, the expressions of CircFAT1, miR-296-3p and MAPRE1 were detected with real-time fluorescent quantitative PCR and Western blotting. The proliferation and apoptosis of CNE2 cells in each group were detected with CCK-8 assay, 5-Ethynyl-2’-deoxyuridine_(Edu)staining and flow cytometry. CNE2-RR cells cultured in vitro were randomly divided into the control group, radiation group, radiation+negative control group, radiation+CircFAT1 knockdown group, and radiation+CircFAT1 knockdown+miR-296-3p inhibitor group. After transfection, the expressions of CircFAT1, miR-296-3p and MAPRE1 were detected with real-time fluorescent quantitative PCR and Western boltting; the proliferation and apoptosis of CNE2-RR cells in each group were detected with CCK-8 assay, Edu staining and flow cytometry. The targeting regulation of CircFAT1 on miR-296-3p and miR-296-3p on MAPRE1 were detected with double luciferase reporter gene assay. Results Compared with NP69 cells, CNE2 and CNE2-RR cells had increased expression of CircFAT1 and mRNA and protein expressions of MAPRE1(P<0.05), but decreased expression of miR-296-3p(P<0.05). Compared with CNE2 cells, CNE2-RR cells had increased mRNA and protein expressions of CircFAT1 and MAPRE1(P<0.05), but decreased expression of miR-296-3p(P<0.05). Compared with the control group, the CircFAT1 knockdown group had decreased mRNA and protein expressions of CircFAT1 and MAPRE1, and reduced survival rate and proliferation rate(P<0.05), but increased expression of miR-296-3p and apoptosis rate(P<0.05). There were no significant changes in the indexes of the negative control group(P>0.05); miR-296-3p inhibitor reversed the effects of CircFAT1 knockdown on the indexes. Compared with the control group and radiation group, the radiation + CircFAT1 knockdown group had decreased mRNA and protein expressions of CircFAT1 and MAPRE1, and reduced survival rate and proliferation rate(P<0.05), but increased expression of miR-296-3p and apoptosis rate(P<0.05). There were no significant changes in the cell indexes of the radiation group and radiation + negative control group(P>0.05); miR-296-3p inhibitor reversed the effects of CircFAT1 knockdown on the indicators of CNE2-RR cells under radiation treatment. CircFAT1 targeted and downregulated the expression of miR-296-3p, and miR-296-3p targeted and downregulated the expression of MAPRE1 in CNE2-RR cells. Conclusion Knockdown of CircFAT1 can reduce the expression of MAPRE1 by upregulating miR-296-3p, thereby inhibiting the proliferation, promoting the apoptosis, and enhancing the radiosensitivity of nasopharyngeal carcinoma cells.

Key words: Circular RNA fat atypical cadherin 1, Microtubule-associated protein, Nasopharyngeal carcinoma, Proliferation, Apoptosis, Radiosensitivity

中图分类号: 

  • R739.63
[1] King AD. MR Imaging of nasopharyngeal carcinoma [J]. Magn Reson Imaging Clin N Am, 2022, 30(1): 19-33.
[2] Ng WT, Chow JCH, Beitler JJ, et al. Current radiotherapy considerations for nasopharyngeal carcinoma [J]. Cancers(Basel), 2022, 14(23): 5773-5787.
[3] Wei H, Yan S, Hui Y, et al. CircFAT1 promotes hepatocellular carcinoma progression via miR-30a-5p/REEP3 pathway [J]. J Cell Mol Med, 2020, 24(24): 14561-14570.
[4] Hu B, Xian Z, Zou Q, et al. CircFAT1 suppresses colorectal cancer development through regulating miR-520b/ UHRF1 axis or miR-302c-3p/ UHRF1 axis [J]. Cancer Biother Radiopharm, 2021, 36(1): 45-57.
[5] Jia L, Wang Y, Wang CY. circFAT1 promotes cancer stemness and immune evasion by promoting STAT3 activation [J]. Adv Sci(Weinh), 2021, 8(13): 2003376-2003391.
[6] Shi F, Yang Q, Shen D, et al. CircRNA WHSC1 promotes non-small cell lung cancer progression via sponging microRNA-296-3p and up-regulating expression of AKT serine/threonine kinase 3 [J]. J Clin Lab Anal, 2021, 35(8): e23865-e23873.
[7] Chu J, Tao L, Yao T, et al. Circular RNA circRUNX1 promotes papillary thyroid cancer progression and metastasis by sponging MiR-296-3p and regulating DDHD2 expression [J]. Cell Death Dis, 2021, 12(1): 112-127.
[8] Deng X, Liu Z, Liu X, et al. miR-296-3p negatively regulated by nicotine stimulates cytoplasmic translocation of c-Myc via MK2 to suppress chemotherapy resistance [J]. Mol Ther, 2018, 26(4): 1066-1081.
[9] Lim Y, Gang DY, Lee WY, et al. Proteomic identification of arginine-methylated proteins in colon cancer cells and comparison of messenger RNA expression between colorectal cancer and adjacent normal tissues [J]. Ann Coloproctol, 2022, 38(1): 60-68.
[10] Tang L, Xiong W, Zhang L, et al. CircSETD3 regulates MAPRE1 through miR-615-5p and miR-1538 sponges to promote migration and invasion in nasopharyngeal carcinoma [J]. Oncogene, 2021, 40(2): 307-321.
[11] Zhang L, Peng H, Xu Z, et al. Circular RNA SOX13 promotes malignant behavior and cisplatin resistance in non-small cell lung cancer through targeting microRNA-3194-3p/microtubule-associated protein RP/EB family member 1[J]. Bioengineered, 2022, 13(1):1814-1827.
[12] 张荣芳, 王春玲, 李申, 等. 甘草素通过影响鼻咽癌CNE-2细胞的自噬增强其放疗敏感性[J]. 中国肿瘤生物治疗杂志, 2018, 25(6): 601-606. ZHANG Rongfang, WANG Chunling, LI Shen, et al. Licorice enhances radio-sensitivity of nasopharyngeal carcinoma CNE-2 cells by affecting autophagy[J]. Chinese Journal of Cancer Biotherapy, 2018, 25(6):601-606.
[13] Wang C, Chen J, Su L, et al. The psychological status in patients with nasopharyngeal carcinoma during radiotherapy [J]. Eur Arch Otorhinolaryngol, 2022, 279(2): 1035-1042.
[14] Li J, Li Y, Liu L, et al. LINC01140 regulates radiosensitivity of nasopharyngeal carcinoma cells through the ceRNA network [J]. Cell Mol Biol(Noisy-le-grand), 2022, 68(4): 75-85.
[15] Zhou B, Li T, Xie R, et al. CircFAT1 facilitates cervical cancer malignant progression by regulating ERK1/2 and p38 MAPK pathway through miR-409-3p/CDK8 axis [J]. Drug Dev Res, 2021, 82(8): 1131-1143.
[16] 刘士标, 张淑君, 李培龙, 等. cg20657709位点甲基化对肺腺癌早期诊断的初步探讨[J].山东大学学报(医学版), 2023, 61(4): 18-25. LIU Shibiao, ZHANG Shujun, LI Peilong, et al. The cg20657709 site methylation in the early detection of lung adenocarcinoma[J]. Journal of Shandong University(Health Sciences), 2023,61(4):18-25.
[17] Li C, Liu Y, Lv Z, et al. Circular RNA circHECTD1 facilitates glioma progression by regulating the miR-296-3p/SLC10A7 axis [J]. J Cell Physiol, 2021, 236(8): 5953-5965.
[18] Tian D, Luo L, Wang T, et al. MiR-296-3p inhibits cell proliferation by the SOX4-Wnt/βcatenin pathway in triple-negative breast cancer [J]. J Biosci, 2021, 46: 98-113. doi:10.1007/s 12038-021-00219-6.
[19] Rodrigues-Ferreira S, Nehlig A, Monchecourt C, et al. Combinatorial expression of microtubule-associated EB1 and ATIP3 biomarkers improves breast cancer prognosis [J]. Breast Cancer Res Treat, 2019, 173(3):573-583.
[20] Liang XH, Feng ZP, Liu FQ, et al. MAPRE1 promotes cell cycle progression of hepatocellular carcinoma cells by interacting with CDK2 [J]. Cell Biol Int, 2020, 44(11): 2326-2333.
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