[1]刘 伟a,周 杨a,边 超a,等.miR-495 对食管癌细胞株Eca109 在不同放射剂量和顺铂浓度作用的影响及机制研究[J].现代检验医学杂志,2022,37(04):13-17+27.[doi:10.3969/j.issn.1671-7414.2022.04.003]
　LIU Weia,ZHOU Yanga,BIAN Chaoa,et al.Effect of miR-495 on Esophageal Cancer Cell Line Eca109 at Different Radiation Doses and Cisplatin Concentrations and Its Mechanism[J].Journal of Modern Laboratory Medicine,2022,37(04):13-17+27.[doi:10.3969/j.issn.1671-7414.2022.04.003]
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miR-495 对食管癌细胞株Eca109 在不同放射剂量和顺铂浓度作用的影响及机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年04期
页码:
13-17+27
栏目:
论著
出版日期:
2022-07-15

文章信息/Info

Title:
Effect of miR-495 on Esophageal Cancer Cell Line Eca109 at Different Radiation Doses and Cisplatin Concentrations and Its Mechanism
文章编号:
1671-7414（2022）04-013-06
作者:
刘 伟a;  周 杨a;  边 超a;  东 丽b
内蒙古自治区人民医院a. 放射治疗科；b. 肿瘤内科, 呼和浩特 010010
Author(s):
LIU Weia;  ZHOU Yanga;  BIAN Chaoa;  DONG lib
a.Department of Radiotherapy；b. Department of Oncology, Inner Mongolia People’s Hospital, Hohhot 010010,China
关键词:
食管癌; 微小核糖核酸-495; 凋亡; 放化疗敏感度
分类号:
R735.1；R730.43
DOI:
10.3969/j.issn.1671-7414.2022.04.003
文献标志码:
A
摘要:
目的 探讨微小核糖核酸(microRNA,miR) -495 对食管癌细胞株Eca109 在不同放射剂量和顺铂浓度作用下的影响及机制。方法 采用分次放疗递增法诱导建立放射抵抗型细胞株Eca109（Eca109-RAD）及分次顺铂递增法诱导建立顺铂耐药型细胞株Eca109（Eca109-DDP），实时荧光定量聚合酶链反应 (qRT-PCR) 检测miR-495 在Eca109-RAD 及Eca109-DDP 细胞中的表达。Eca109-RAD 及Eca109-DDP 分为NC 组和miR-495 mimic 组，转染后qRT-PCR 检测各组细胞中miR-495 的表达，CCK8 检测不同放射剂量对Eca109-RAD 细胞和Eca109 细胞存活能力的影响，及不同浓度的顺铂对Eca109-DDP 细胞和Eca109 细胞存活能力的影响，NC 组和miR-495 mimic 组经放射及顺铂处理后，采用流式细胞仪检测各组细胞的凋亡率，Western blot 检测各组细胞中凋亡相关蛋白caspase 3,Bax 和Bcl-2 蛋白的表达。结果 与Eca109 细胞（0.99±0.01）相比，Eca109-RAD 细胞中miR-495 的表达（0.48±0.03 ）及Eca109-DDP 细胞中miR-495的表达（0.52±0.05）均降低，差异有统计学意义(t=27.930，15.970, 均P=0.000)。与NC 组细胞相比，miR-495 mimic组Eca109-RAD 细胞中miR-495 的表达（4.82±0.48 vs 1.00±0.03）及Eca109-DDP 细胞中miR-495 的表达（5.68±0.54vs 1.00±0.01）均显著增加，差异有统计学意义(t=13.760，15.100, 均P=0.000)。与NC 组相比，miR-495 mimic 组Eca109-RAD 细胞对放疗敏感度增加，差异均有统计学意义(t=6.780 ～ 18.860，P = 0.000 ～ 0.001)；与NC 组相比，miR-495 mimic 组Eca109-DDP 细胞对顺铂敏感度增加，差异均有统计学意义(t=7.510 ～ 21.630，均P=0.000)。经放射处理后，与NC 组相比，miR-495 mimic 组Eca109-RAD 细胞平板克隆形成能力（46.33±5.69 个vs 93.33±4.51 个）及Eca109-DDP（34.67±2.52 个vs 89.00±4.03 个）细胞平板克隆形成能力显著降低(t=11.210，19.800, 均P=0.000) 。经放射处理后，与NC 组相比，miR-495 mimic 组Eca109-RAD 细胞凋亡率（25.66%±2.41% vs 8.39%±0.82%）及Eca109-DDP 细胞凋亡率（21.05%±5.37% vs 8.67%±1.15%）均显著增加(t=11.750，10.030，P=0.000，0.001)。经放射处理后，与NC 组相比，miR-495 mimic 组Eca109-RAD 细胞及Eca109-DDP 细胞中Bcl-2 蛋白表达均降低(t=4.650，7.450，P=0.006，0.001)，Bax 和caspase 3 蛋白的表达均增加(t= 7.100 ～ 14.290, P=0.000 ～ 0.001)。结论 MiR-495 可能通过调控凋亡相关蛋白促进食管癌细胞放化疗敏感度。
Abstract:
Objective To investigate the effect and mechanism of microRNA (miR)-495 on esophageal cancer cell line Eca109 under different radiation doses and cisplatin concentrations.Methods The radiation resistant cell line Eca109 (Eca109 RAD) was induced by fractionated radiotherapy and the cisplatin resistant cell line Eca109 (Eca109 DDP) was induced by fractionated cisplatin,the expression of miR-495 in Eca109-RAD and Eca109-DDP cells was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR).Eca109-RAD and Eca109-DDP were divided into NC group and miR-495 mimic group. After transfection, the expression of miR-495 was detected by qRT-PCR,CCK8 was used to detect the effects of different radiation doses on the viability of Eca109 RAD cells and Eca109 cells, and the effects of different concentrations of cisplatin on the viability of Eca109 DDP cells and Eca109 cells.After radiation and cisplatin treatment, the apoptosis rate of cell in NC group and miR-495 mimic group was detected by flow cytometry, and the expressions of apoptosis related proteins caspase 3, Bax and Bcl-2 in each group were detected by Western blot. Results Compared with Eca109 cells(0.99 ± 0.01), the expression of miR-495 in Eca109-RAD cells (0.48 ± 0.03) and Eca109-DDP cells (0.52 ± 0.05)decreased ，the differences were statistically significant(t = 27.930，15.970, all P = 0.000). Compared with NC group, the expression of miR-495 in Eca109-RAD cells (1.00 ± 0.03 vs 4.82 ± 0.48) and Eca109-DDP cells (1.00 ± 0.01 vs 5.68 ± 0.54) in miR-495 mimic group were significantly increased ，the differences were statistically significant (t=13.760，15.100, all P = 0.000).Compared with NC group, Eca109-RAD cells in miR-495 mimic group were more sensitive to radiotherapy，the differences were statistically significant (t=6.780 ～ 18.860, P=0.000 ～ 0.0010 ）,and compared with NC group, Eca109-DDP cells in miR-495 mimic group were more sensitive to cisplatin, the differences were statistically significant (t=7.510 ～ 21.630, all P= 0.000). After radiation treatment, compared with NC group, the plate clone forming ability of Eca109-RAD cells (93.33 ± 4.51 vs 46.33 ± 5.69) and Eca109-DDP cells (89.00 ± 4.03 vs 34.67 ± 2.52) in miR-495 mimic group decreased significantly, the differences were statistically significant (t =11.210，19.800, all P = 0.000).After radiation treatment, compared with NC group, the apoptosis rates of Eca109-RAD cells (8.39%± 0.82% vs 25.66%± 2.41%) and Eca109-DDP cells (8.67%±1.15% vs 21.05%± 5.37%) in miR-495 mimic group increased significantly (t = 11.750，10.030, P=0.000，0.001). After radiation treatment, compared with NC group, the expression of Bcl-2 protein，0.001), and the expression of Bax and caspase 3 protein increased (t= 7.100 ～ 14.290, P=0.000 ～ 0.001). Conclusion MiR-495 may promote the chemoradiotherapy sensitivity of esophageal cancer cells by regulating apoptosis related proteins.

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备注/Memo

备注/Memo:
基金项目：内蒙古自治区自然科学基金（2019MS08090）。
作者简介：刘伟（1979-），男，硕士研究生，副主任医师，研究方向: 肿瘤的放射治疗及综合治疗，E-mail:liuwei1979703@163.com。
通讯作者：周杨（1987-），男，主治医师，E-mail:240123137@qq.com。
更新日期/Last Update: 1900-01-01