LncRNA MIR503HG通过调控miR-224-5pCHSY1表达对结肠癌细胞放射敏感性的影响  

Study on the effects of LncRNA MIR503HG on radiosensitivity of colon cancer cells by regulating the expression of miR-224-5pCHSY1

作　　者：张健 李文君 Zhang Jian;Li Wenjun(Department of Radiotherapy,PLA Rocket Force Characteristic Medical Center,Beijing 100088,China)

机构地区：[1]火箭军特色医学中心放射治疗科,北京100088

出　　处：《国际放射医学核医学杂志》2023年第11期674-689,共16页International Journal of Radiation Medicine and Nuclear Medicine

摘　　要：目的探讨长链非编码RNA微小RNA-503宿主基因(LncRNA MIR503HG)通过调控miRNA-224-5p硫酸软骨素合酶1 (miR-224-5pCHSY1 )的表达对结肠癌细胞放射敏感性的影响。方法选取2019年3月至2022年1月火箭军特色医学中心收治的48例结肠癌患者为组织样本获取对象, 进行回顾性分析, 其中男性28例、女性20例, 年龄(57.43±5.28)岁。根据放疗后的病灶情况将患者分为放射抵抗组(23例)和放射敏感组(25例)。采用实时定量聚合酶链式反应(qRT-PCR)检测2组患者结肠癌组织及各细胞株(CCD841、COLO320、SW480、RKO、HCT116)中LncRNA MIR503HG、miR-224-5pCHSY1的表达情况;构建放射抵抗的结肠癌细胞株HCT116R, 将HCT116R细胞株分为过表达LncRNA MIR503HG(MIR503HG)组及其阴性对照组(mimiC-NC)、miR-224-5pCHSY1抑制组(anti-miR-224-5p)及其阴性对照组(anti-miR-NC)、过表达LncRNA MIR503HG+过表达miR-224-5pCHSY1组(MIR503HG+miR-224-5p)、过表达LncRNA MIR503HG+阴性对照组(MIR503HG+miR-NC);通过双荧光素酶报告验证LncRNA MIR503HG与miR-224-5pCHSY1的靶向作用;检测各组细胞的存活分数(SF)和凋亡率。两组间数据的比较使用t检验。结果与放射敏感组相比, 放射抵抗组结肠癌组织中LncRNA MIR503HG的相对表达量明显降低(1.40±0.36对0.72±0.17), miR-224-5pCHSY1的相对表达量明显升高(1.06±0.25对1.54±0.27 ), 且差异均有统计学意义(t=8.247、6.529, 均P<0.05)。CCD841、COLO320、SW480、RKO、HCT116及HCT116R各细胞株LncRNA MIR503HG的相对表达量分别为2.38±0.06、1.03±0.05、0.87±0.03、0.86±0.02、0.77±0.04、0.54±0.09, miR-224-5pCHSY1的相对表达量分别为0.38±0.06、0.56±0.01、0.59±0.02、0.59±0.05、0.63±0.04、0.82± 0.06, 与正常细胞株CCD841相比, 结肠癌细胞株COLO320、SW480、RKO、HCT116的LncRNA MIR503HG相对表达量均明显降低(t=2.061、1.665、4.058、6.201, 均P<0.05), miR-224-5pCHSY1的相对表达量均明显增加(t=1.238、1.930、2.037、1.742, 均P<0.05 )。�Objective To investigate the effects of long non-coding RNA microRNA503 host gene(LncRNA MIR503HG)on the radiosensitivity of colon cancer cells by regulating the expression of microRNA-224-5p chondroitin sulfate synthas 1(miR-224-5pCHSY1).Methods A retrospective analysis was conducted on 48 patients with colon cancer treated in PLA Rocket Force Characteristic Medical Center from March 2019 to January 2022.These patients were selected as tissue samples.The cohort included 28 males and 20 females,aged(57.43±5.28)years.On the basis of the lesions after radiotherapy,they were divided into the radiation resistance group(n=23)and the radiosensitive group(n=25).The expression levels of LncRNA MIR503HG and miR-224-5pCHSY1 in colon cancer tissues and cell lines(CCD841,COLO320,SW480,RKO,and HCT116)were detected by real-time quantitative polymerase chain reaction(qRT-PCR).The radiation-resistant colon cancer cell line HCT116R was constructed and divided into the overexpressing LncRNA MIR503HG(MIR503HG)and its negative control group(mimic-NC),miR-224-5pCHSY1 inhibition group(anti-miR-224-5p)and its negative control group(anti-miR-NC),overexpressing LncRNA MIR503HG+overexpressing miR-224-5pCHSY1 group(MIR503HG+miR-224-5p),and overexpressing LncRNA MIR503HG+negative control group(MIR503HG+miR-NC).The targeting effect of LncRNA MIR503HG and miR-224-5pCHSY1 was verified by dual-luciferase assay,and the cell survival fraction(SF)and apoptosis rate were detected.The data between two groups were compared using t-test.Results Compared with the radiosensitive group,the expression of LncRNA MIR503HG in the radiation resistance group was significantly lower(1.40±0.36 vs.0.72±0.17),and the expression of miR-224-5pCHSY1 was significantly higher(1.06±0.25 vs.1.54±0.27)in the radioresistant group,and the differences were statistically significant(t=8.247,6.529;both P<0.05).The relative expression levels of LncRNA MIR503HG in the CCD841,COLO320,SW480,RKO,HCT116,and HCT116R cell lines were 2.38±0.06,1.03±0.05,0.87±0.03,0.86±0.02,0.7

关 键 词：结肠肿瘤 辐射耐受性 RNA 长链非编码 LncRNA MIR503HG miR-224-5pCHSY1 

分 类 号：R735.35[医药卫生—肿瘤]