Effect of upregulated HuR gene on radiosensitivity of esophageal squamous cell carcinoma cell Kyse450
Han Dan1, Li Lu1, Kan Zhiwen2, Tao Zhenchao3, Qian Liting3
1Department of Life Science and Medicine, University of Science and Technology of China,Hefei 230026,China; 2Anhui Medical University Affiliated Provincial Hospital,Hefei 230022,China; 3Department of Radiotherapy,West District of the First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Cancer Hospital),Hefei 230031,China
Abstract：Objective To evaluate the effect of upregulation of HuR gene on the radiosensitivity of esophageal squamous cell carcinoma cell Kyse450. Methods The HuR gene of Kyse450 cells was upregulated by lentivirus. At the same time, X-ray irradiation at a dose of 6Gy was selected as the intervention condition. Western blot and qPCR were used to detect the expression levels of protein and RNA after Kyse450 transfection, respectively. CCK8 kit was employed to determine the cell proliferation rate. Clone formation assay was adopted to evaluate the ability of cell clone formation. Wound healing experiment and the Transwell test were performed to detect changes in cell migration. Results CCK8 assay showed that the proliferation ability of cells was enhanced after upregulation of HuR gene, and this enhancement trend was more obvious after radiation. The plate cloning experiment showed that with the increase of radiation dose, the clone formation rates were decreased in both groups, but the clone formation rate in the overexpression group was higher than that in the control group. Wound healing experiment and Transwell test demonstrated that the wound healing rate and migration ability in the overexpression group were higher than those in the control group, and the difference was more significant after radiotherapy. Western blot showed that the levels of MMP9 and MMP2 at 24h after radiotherapy in the overexpression group were higher than those in the control group. Conclusion The upregulation of HuR can enhance the proliferation, cloning, migration capabilities and decrease the radiosensitivity of esophageal squamous cell carcinoma cells.
Han Dan,Li Lu,Kan Zhiwen et al. Effect of upregulated HuR gene on radiosensitivity of esophageal squamous cell carcinoma cell Kyse450[J]. Chinese Journal of Radiation Oncology, 2022, 31(5): 456-461.
[1] Imazeki H, Kato K. Development of chemotherapeutics for unresectable advanced esophageal cancer[J]. Expert Rev Anticancer Ther, 2020, 20(12):1083-1092. DOI:10.1080/14737140.2020.1814149. [2] 丁秀平,李宝生. 从胸段食管癌淋巴结转移规律探讨单纯放疗靶区的定义[J]. 中华放射肿瘤学杂志,2012,21(1):34-37. DOI:10.3760/cma.j.issn.1004-4221.2012.01.011. Ding XP, Li BS. From the law of thoracic segment esophageal cancer lymph node metastasis to explore simple definition for target of radiotherapy[J]. Chin J Radiat Oncol, 2012,21(1):34-37. DOI:10.3760/cma.j.issn.1004-4221.2012.01.011. [3] Song X, Shi X, Li W, et al. The RNA-binding protein HuR in digestive system tumors[J]. Biomed Res Int, 2020, 2020:9656051. DOI:10.1155/2020/9656051. [4] Xu X, Song C, Chen Z, et al. Downregulation of HuR inhibits the progression of esophageal cancer through interleukin-18[J]. Cancer Res Treat, 2018, 50(1):71-87. DOI:10.4143/crt.2017.013. [5] Wang A, Bao Y, Wu Z, et al. Long noncoding RNA EGFR-AS1 promotes cell growth and metastasis via affecting HuR mediated mRNA stability of EGFR in renal cancer[J]. Cell Death Dis, 2019, 10(3):154. DOI:10.1038/s41419-019-1331-9. [6] Xu J, Yang B, Wang L, et al. LncRNA BBOX1-AS1 upregulates HOXC6 expression through miR-361-3p and HuR to drive cervical cancer progression[J]. Cell Prolif, 2020, 53(7):e12823. DOI:10.1111/cpr.12823. [7] Zhang F, Cai Z, Lv H, et al. Multiple functions of HuR in urinary tumors[J]. J Cancer Res Clin Oncol, 2019, 145(1):11-18. DOI:10.1007/s00432-018-2778-2. [8] Filippova N, Yang X, Wang Y, et al. The RNA-binding protein HuR promotes glioma growth and treatment resistance[J]. Mol Cancer Res, 2011, 9(5):648-659. DOI:10.1158/1541-7786.MCR-10-0325. [9] Richards NG, Rittenhouse DW, Freydin B, et al. HuR status is a powerful marker for prognosis and response to gemcitabine-based chemotherapy for resected pancreatic ductal adenocarcinoma patients[J]. Ann Surg, 2010, 252(3):499-505;discussion 505-506. DOI:10.1097/SLA.0b013e3181f1fd44. [10] Mehta M, Basalingappa K, Griffith JN, et al. HuR silencing elicits oxidative stress and DNA damage and sensitizes human triple-negative breast cancer cells to radiotherapy[J]. Oncotarget, 2016, 7(40):64820-64835. DOI:10.18632/oncotarget.11706. [11] Papatheofani V, Levidou G, Sarantis P, et al. HuR protein in hepatocellular carcinoma:implications in development, prognosis and treatment[J]. Biomedicines, 2021, 9(2):119. DOI:10.3390/biomedicines9020119. [12] Milne AN, Carvalho R, Morsink FM, et al. Early-onset gastric cancers have a different molecular expression profile than conventional gastric cancers[J]. Mod Pathol, 2006, 19(4):564-572. DOI:10.1038/modpathol.3800563. [13] Wu M, Tong C, Yan W, et al. The RNA binding protein HuR:A promising drug target for anticancer therapy[J]. Curr Cancer Drug Targets, 2019, 19(5):382-399. DOI:10.2174/1568009618666181031145953. [14] Richard V, Kumar T, Pillai RM. Transitional dynamics of cancer stem cells in invasion and metastasis[J]. Transl Oncol, 2021, 14(1):100909. DOI:10.1016/j.tranon.2020.100909. [15] Xiong W, Liao Y, Qin JY, et al. Adverse effects of chemoradiotherapy on invasion and metastasis of tumor cells[J]. Genes Dis, 2020, 7(3):351-358. DOI:10.1016/j.gendis.2020.04.004. [16] Dziembowska M, Wlodarczyk J. MMP9:a novel function in synaptic plasticity[J]. Int J Biochem Cell Biol, 2012, 44(5):709-713. DOI:10.1016/j.biocel.2012.01.023.
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