[1]李智军a,姜海燕b,边 超a,等.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[J].现代检验医学杂志,2023,38(02):25-31.[doi:10.3969/j.issn.1671-7414.2023.02.005 ]
　LI Zhi-juna,JIANG Hai-yanb,BIAN Chaoa,et al.Mechanism of LINC01503 Promotes Radiotherapy Resistance to Esophageal Squamous Cell Carcinoma by Regulating ERK Phosphate Acidization[J].Journal of Modern Laboratory Medicine,2023,38(02):25-31.[doi:10.3969/j.issn.1671-7414.2023.02.005 ]
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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()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年02期
页码:
25-31
栏目:
论著
出版日期:
2023-03-15

文章信息/Info

Title:
Mechanism of LINC01503 Promotes Radiotherapy Resistance to Esophageal Squamous Cell Carcinoma by Regulating ERK Phosphate Acidization
文章编号:
1671-7414（2023）02-025-07
作者:
李智军a; 姜海燕b; 边 超a; 张浩伟a; 张斯琴胡a
（内蒙古自治区人民医院 a. 放射治疗科；b. 教学处，呼和浩特 010017）
Author(s):
LI Zhi-juna; JIANG Hai-yanb; BIAN Chaoa; ZHANG Hao-weia; ZHANG Si-qin-hua
（a. Department of Radiotherapy；b. Department of Teaching，People’s Hospital of Inner Mongolia Autonomous Region，Hohhot 010017，China）
关键词:
食管鳞状细胞癌; LINC01503; 放疗抵抗; 增殖; 凋亡
分类号:
R735.1；R730.43
DOI:
10.3969/j.issn.1671-7414.2023.02.005
文献标志码:
A
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（χ2=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 LINC01503通过促进 ERK磷酸化调控细胞周期和凋亡促进 ESCC细胞放疗抵抗，是逆转 ESCC放疗抵抗的潜在分子靶点。
Abstract:
Objective 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 (χ2=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 P<0.001). In si-LINC01503 group, KYSE150R cell cycle was arrested in G0/G1 phase (61.47% ± 3.60% vs 52.15% ± 2.11%), apoptosis increased (31.95%±2.40% vs 3.68% ± 0.47%), the differences were statistically significant (t=4.602, 20.022, P=0.004, 0.002), respectively. 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), Bcl-2 (0.16 ± 0.03 vs 0.85 ± 0.07) protein expression decreased, while Bax protein expression increased (0.69 ± 0.06 vs 0.22 ± 0.05), the differences were statistically significant (t=4.718～18.440, all P<0.05).Conclusion LINC01503 is a potential molecular target to reverse ESCC radioresistance by promoting ERK phosphorylation, regulating cell cycle and apoptosis, and promoting ESCC radioresistance.

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备注/Memo

备注/Memo:
收稿日期：2022-09-06修回日期：2022-11-30
基金项目：内蒙古自治区科学技术厅科研项目（编号：2018MS0115）。
作者简介：李智军（1975-），男，博士，主任医师，研究方向：食管癌放射治疗， E-mail：zhilijun5@163.com。
通讯作者：姜海燕（1982-），女，硕士研究生，研究方向：放射治疗教学工作。

更新日期/Last Update: 2023-03-15