LncRNA LINC00261 regulates radiosensitivity of nasopharyngeal carcinoma and tumor formation in nude mice by down-regulating miR-620 expression
Fu Gaoshang1, Zhang Ke1, Xu Yanxia1, Xu Ying1, Zhang Xuexi2, Lian Lixia3
1Department of Otolaryngology Head and Neck Surgery, Children’s Hospital Affiliated to Zhengzhou University, Henan Children's Hospital, Zhengzhou 450000, China; 2Department of Otolaryngology Head andNeck Surgery, Beijing Children's Hospital, Capital Medical University, Beijing 100045, China; 3Department of Cancer Medical Center, Henan Provincial People's Hospital, Zhengzhou 450003, China
Abstract:Objective To investigate the effect of LINC00261 on the radiosensitivity of nasopharyngeal carcinoma and tumor formation and its underlying mechanism in nude mice. Methods qRT-PCR was used to detect the relative expression levels of miR-620 and LINC00261 in radiosensitive and radioresistant nasopharyngeal carcinoma tissues. After the 6-10B and HNE-3 cells were irradiated with 0, 2, 4, 6, and 8 Gy 60Coγ-ray, the relative expression levels of miR-620 and LINC00261 were measured by qRT-PCR. After over-expression or silencing of LINC00261 and inhibition of miR-620 expression, the cells were irradiated with 4 Gy 60Coγ-ray. Clone formation assay was performed to detect the radiosensitivity of nasopharyngeal carcinoma cells. Flow cytometry was used to detect cell apoptosis. Western blot was utilized to detect the expression levels of Cleaved caspase-3 and Cleaved caspase-9 proteins. Luciferase reporter assay was adopted to analyze the targeting relationship between LINC00261 and miR-620. The changes in tumor formation were observed in tumor-bearing nude mice. Results Compared with the radiosensitive tissues, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in radioresistant tissues (both P<0.05). After different doses of irradiation, the expression of LINC00261 was significantly down-regulated, whereas that of miR-620 was significantly up-regulated in 6-10B and HNE-3 cells (both P<0.05). After overexpression of LINC00261 and interference with miR-620 expression, the expression levels of Cleaved caspase-3 and Cleaved caspase-9 were significantly up-regulated (both P<0.05), the cell apoptosis rate was remarkably increased (P<0.05) and the cell survival fraction was significantly enhanced in 6-10B and HNE-3 cells (P<0.05). LINC00261 targetedly regulated the expression of miR-620. Overexpression of miR-620 could attenuate the radiosensitization and pro-apoptotic effects of LINC00261 overexpression on nasopharyngeal carcinoma cells. LINC00261 overexpression could significantly reduce the tumor formation weight of nasopharyngeal carcinoma in nude mice (P<0.05). Conclusion Overexpression of LINC00261 can increase the radiosensitivity of nasopharyngeal carcinoma cells probably by targeted regulation of miR-620 expression.
Fu Gaoshang,Zhang Ke,Xu Yanxia et al. LncRNA LINC00261 regulates radiosensitivity of nasopharyngeal carcinoma and tumor formation in nude mice by down-regulating miR-620 expression[J]. Chinese Journal of Radiation Oncology, 2021, 30(2): 198-203.
[1] 陆海军,刘霁,丁晓. 鼻咽癌的综合治疗研究进展[J]. 山东大学耳鼻喉眼学报, 2019, 33(2):26-30. DOI:10.6040/j.issn.1673-3770.1.2019.010. Lu HJ, Liu J, Ding X. Progress in the comprehensive treatment for nasopharyngeal carcinoma[J]. J Otolaryngol Ophthalmol Shandong Uni, 2019, 33(2):26-30. DOI:10.6040/j.issn.1673-3770.1.2019.010. [2] 陈瑶, 李思维. 鼻咽癌放射敏感性的研究进展[J]. 华夏医学, 2017, 30(1):167-171. DOI:10.19296/j.cnki.1008-2409.2017-01-053. Chen Y, Li SW. Research progress on radiosensitivity of nasopharyngeal carcinoma[J]. China Med, 2017, 30(1):167-171. DOI:10.19296/j.cnki.1008-2409.2017-01-053. [3] 罗文娜, 罗伟濠, 罗迪贤. 鼻咽癌相关长链非编码RNAs研究进展[J]. 山东医药, 2018, 58(42):112-115. DOI:10.3969/j.issn.1002-266X.2018.42.033. Luo WN, Luo WH, Luo DX. Research progress on long non-coding RNAs related to nasopharyngeal carcinoma[J]. Shandong Med J, 2018, 58(42):112-115. DOI:10.3969/j.issn.1002-266X.2018.42.033. [4] Shahabi S, Kumaran V, Castillo J, et al. LINC00261 is an epigenetically regulated tumor suppressor essential for activation of the DNA damage response[J]. Cancer Res, 2019, 79(12):3050-3062. DOI:10.1158/0008-5472. CAN-18-2034. [5] Wang ZK, Yang L, Wu LL, et al. Long non-coding RNA LINC00261 sensitizes human colon cancer cells to cisplatin therapy[J]. Braz J Med Biol Res, 2018, 51(2):e6793. DOI:10.1590/1414-431X20176793. [6] Lin K, Jiang H, Zhuang SS, et al. Long noncoding RNA LINC00261 induces chemosensitization to 5-fluorouracil by mediating methylation-dependent repression of DPYD in human esophageal cancer[J]. FASEB J, 2019, 33(2):1972-1988. DOI:10.1096/fj.201800759R. [7] 梅雪霜, 胡洪义, 田怀谷, 等. 抑制miR-210的表达对鼻咽癌放射抵抗细胞敏感性的影响[J]. 中国老年学杂志, 2019, 39(07):1669-1672. DOI:10.3969/j.issn.1005-9202.2019.07.046. [8] Huang X, Taeb S, Jahangiri S, et al. miR-620 promotes tumor radioresistance by targeting 15-hydroxyprostaglandin dehydrogenase (HPGD)[J]. Oncotarget, 2015, 6(26):22439-22451. DOI:10.18632/oncotarget.4210. [9] Wu C, Zhao A, Tan T, et al. Overexpression of microRNA-620 facilitates the resistance of triple negative breast cancer cells to gemcitabine treatment by targeting DCTD[J]. Exp Ther Med, 2019, 18(1):550-558. DOI:10.3892/etm.2019.7601. [10] 周苏娜, 张明鑫, 张琰君, 等. miRNA-381表达及其与鼻咽癌放射敏感性的关系[J]. 现代肿瘤医学, 2017, 25(19):22-26. DOI:10.3969/j.issn.1672-4992.2017.19.004. [11] Shi J, Ma H, Wang H, et al. Overexpression of LINC00261 inhibits non-small cell lung cancer cells progression by interacting with miR-522-3p and suppressing Wnt signaling[J]. J Cell Biochem, 2019, 120(10):18378-18387. DOI:10.1002/jcb.29149. [12] Yan D, Liu W, Liu Y, et al. LINC00261 suppresses human colon cancer progression via sponging miR-324-3p and inactivating the Wnt/β-catenin pathway[J]. J Cell Physiol, 2019, 234(12):22648-22656. DOI:10.1002/jcp.28831. [13] 赵志国, 张力平. miRNA和lncRNA相互作用与恶性肿瘤相关性的研究进展[J]. 现代肿瘤医学, 2018, 26(18):176-179. DOI:10.3969/j.issn.1672-4992.2018.18.040. [14] Li X, Shen M. Circular RNA hsa_circ_103809 suppresses hepatocellular carcinoma proliferation and invasion by sponging miR-620[J]. Eur Rev Med Pharmacol Sci, 2019, 23(2):555-566. DOI:10.26355/eurrev_201902_16868. [15] Tian JDC, Liang L. Involvement of circular RNA SMARCA5/microRNA-620 axis in the regulation of cervical cancer cell proliferation, invasion and migration[J]. Eur Rev Med Pharmacol Sci, 2018, 22(24):8589-8598. DOI:10.26355/eurrev_201812_16622. [16] Zhao Z, Han C, Liu J, et al. GPC5, a tumor suppressor, is regulated by miR-620 in lung adenocarcinoma[J]. Mol Med Rep, 2014, 9(6):2540-2546. DOI:10.3892/mmr.2014.2092.