LINC01503调控ERK磷酸化促进食管鳞状细胞癌放疗抵抗的机制研究  

Mechanism of LINC01503 Promotes Radiotherapy Resistance to Esophageal Squamous Cell Carcinoma by Regulating ERK Phosphate Acidization

作　　者：李智军 姜海燕 边超 张浩伟 张斯琴胡 LI Zhi-jun;JIANG Hai-yan;BIAN Chao;ZHANG Hao-wei;ZHANG Si-qin-hu(Department of Radiotherapy,People’s Hospital of Inner Mongolia Autonomous Region,Hohhot 010017,China;Department of Teaching,People’s Hospital of Inner Mongolia Autonomous Region,Hohhot 010017,China)

机构地区：[1]内蒙古自治区人民医院放射治疗科,呼和浩特010017 [2]内蒙古自治区人民医院教学处,呼和浩特010017

出　　处：《现代检验医学杂志》2023年第2期25-31,共7页Journal of Modern Laboratory Medicine

基　　金：内蒙古自治区科学技术厅科研项目(编号:2018MS0115)。

摘　　要：目的探究长链非编码核糖核酸(LncRNA)LINC01503调控细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)通路磷酸化促进食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)放疗抵抗的作用机制。方法收集2014年6月~2017年12月内蒙古自治区人民医院ESCC患者癌组织及癌旁组织标本79例,采用实时荧光定量PCR检测LINC01503在ESCC组织中的表达水平,分析LINC01503与ESCC患者临床病理参数、放疗敏感性及预后的关系。构建ESCC放疗抵抗细胞株KYSE150R,检测KYSE150R细胞中LINC01503的表达;转染siRNA下调KYSE150R中LINC01503的表达。CCK8实验检测各组细胞放疗敏感性;流式细胞试验检测各组细胞周期和细胞凋亡;Western blot检测各组细胞中ERK1/2,PERK1/2,细胞周期蛋白D1(cyclin D1),细胞周期素依赖性激酶4(cyclin dependent kinase 4,CDK4),凋亡蛋白Bcl-2和Bax蛋白表达。结果与癌旁组织相比,LINC01503在ESCC组织中表达上调(4.15±1.21 vs 0.96±0.43),差异有统计学意义(t=22.083,P<0.001)。LINC01503高表达与患者T分期、淋巴结转移、TNM分期、放疗抵抗有关,差异均有统计学意义(t=2.322~2.939,均P<0.05)。LINC01503预测预后的曲线下面积为0.780(95%CI:0.676~0.884),灵敏度和特异度分别为81.58%,67.05%。LINC01503高表达组5年生存率低于LINC01503低表达组[41.86%(18/43)vs 63.89%(23/36)],差异有统计学意义(χ^(2)=4.430,P=0.035)。与si-NC组相比,si-LINC01503组KYSE150R细胞的放疗敏感性增加,差异均有统计学意义(t=17.391~33.692,均P<0.001);si-LINC01503组KYSE150R细胞周期阻滞在G0/G1期(61.47%±3.60%vs 52.15%±2.11%),细胞凋亡增加(31.95%±2.40%vs 3.68%±0.47%),差异均有统计学意义(t=4.602,20.022;P=0.004,0.002);PERK1/2(0.24±0.03 vs 1.25±0.09),cyclin D1(0.18±0.06 vs 1.40±0.14),CDK4(0.87±0.09 vs 1.37±0.16)和Bcl-2(0.16±0.03 vs 0.85±0.07)蛋白表达降低,Bax蛋白表达增加(0.69±0.06 vs 0.22±0.05),差异均有统计学意义(t=4.718~18.440,均P<0.05)。结论LIObjective To investigate the mechanism of long chain non coding RNA LINC01503 regulating the phosphorylation of extracellular signal regulated kinase(ERK)pathway to promote the radiotherapy resistance of esophageal squamous cell carcinoma(ESCC).Methods 79 cases of cancer tissue and adjacent tissue samples from ESCC patients in People’s Hospital of Inner Mongolia Autonomous Region from June 2014 to December 2017 were collected.The expression level of LINC01503 in ESCC tissue was detected by real-time fluorescent quantitative PCR,and the relationship between LINC01503 and clinical pathological parameters,radiotherapy sensitivity and prognosis of ESCC patients was analyzed.ESCC radiation resistant cell line KYSE150R was constructed,and the expression of LINC01503 in KYSE150R cells was detected.Transfection of siRNA down regulated the expression of LINC01503 in KYSE150R.CCK8 assay was used to detect the radiosensitivity of cells in each group and cell cycle and apoptosis were detected by flow cytometry.Western blot was used to detect the expression of ERK1/2,PERK1/2,cyclin D1,cyclin dependent kinase 4(CDK4),apoptosis protein Bcl-2 and Bax in cells of each group.Results Compared with the adjacent tissues,LINC01,503 was up-regulated in ESCC tissues(4.15±1.21 vs 0.96±0.43),with a statistically significant difference(t=22.083,P<0.001).The overexpression of LINC01503 was related to T stage,lymph node metastasis,TNM stage and radiotherapy resistance,and the differences were statistically significant(t=2.322~2.939,all P<0.05).The area under the curve of LINC01503 predicting prognosis was 0.780(95%CI:0.676~0.884),with sensitivity and specificity of 81.58%and 67.05%,respectively.The 5-year survival rate of LINC01503 overexpression group was lower than that of LINC01503 low expression group[41.86%(18/43)vs 63.88%(23/36)],the difference was statistically significant(χ^(2)=4.430,P=0.035).Compared with the si-NC group,the radiosensitivity of KYSE150R cells in the si-LINC01503 group increased significantly(t=17.391~33.692,all

关 键 词：食管鳞状细胞癌 LINC01503 放疗抵抗 增殖 凋亡 

分 类 号：R735.1[医药卫生—肿瘤] R730.43[医药卫生—临床医学]