[1]罗 文,罗 钢,周玉玲,等.视网膜母细胞瘤患儿血清LncRNA-NEAT1 水平表达及对瘤细胞生物学功能的影响[J].现代检验医学杂志,2024,39(03):115-119+151.[doi:10.3969/j.issn.1671-7414.2024.03.019]
　LUO Wen,LUO Gang,ZHOU Yuling,et al.Expression of Serum LncRNA NEAT1 in Children with Retinoblastoma and Its Impact on the Biological Function of Tumor Cells[J].Journal of Modern Laboratory Medicine,2024,39(03):115-119+151.[doi:10.3969/j.issn.1671-7414.2024.03.019]
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视网膜母细胞瘤患儿血清LncRNA-NEAT1 水平表达及对瘤细胞生物学功能的影响()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年03期
页码:
115-119+151
栏目:
论著
出版日期:
2024-05-15

文章信息/Info

Title:
Expression of Serum LncRNA NEAT1 in Children with Retinoblastoma and Its Impact on the Biological Function of Tumor Cells
文章编号:
1671-7414（2024）03-115-06
作者:
罗 文; 罗 钢; 周玉玲; 胡佩宏; 张青林; 王莎莎; 明 媚
[黄石市中心医院（湖北理工学院附属医院）眼科，湖北黄石 435000]
Author(s):
LUO Wen;  LUO Gang;  ZHOU Yuling;  HU Peihong;  ZHANG Qinglin;  WANG Shasha;  MING Mei
[Department of Ophthalmology, Huangshi Central Hospital（Affiliated Hospital of Hubei Institute of Technology）, Hubei Huangshi 435000, China]
关键词:
视网膜母细胞瘤; 长链非编码RNA 核富集转录体1; 细胞增殖; 细胞生物学特性
分类号:
R739.72；R730.43
DOI:
10.3969/j.issn.1671-7414.2024.03.019
文献标志码:
A
摘要:
目的 探讨长链非编码RNA（long non-coding RNA，LncRNA）核富集转录体1（nuclera-enriched autosomaltranscript，NEAT1）在视网膜母细胞瘤（retinoblastoma，Rb）患儿血清中表达，以及下调Rb 细胞Y79 中NEAT1 对细胞生物学功能的影响。方法 以2015 年3 月～ 2021 年3 月鄂东医疗集团黄石市中心医院诊疗的83 例Rb患儿为研究对象，同期，在儿童保健中心选取健康儿童50 例（对照组），实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）检测血清中NEAT1 表达，分析Rb 患儿和对照组血清NEAT1 表达差异，以及不同临床指标Rb 患者血清中NEAT1 表达差异。培养Y79 细胞并分为si-NEAT1 组（转染NEAT1 的干扰序列）、si-NC 组（转染对照序列）和Ctl 组（仅加入转染试剂），分别使用qRT-PCR，MTT，流式细胞术和Transwell 检测NEAT1 表达、细胞增殖、凋亡、迁移和侵袭情况。结果 Rb 患儿血清中NEAT1 表达量（1.43 ± 0.28）高于对照组（1.01 ± 0.21），差异具有统计学意义（t=9.116，P<0.001）；国际视网膜母细胞瘤分期(Intraocular International Retinoblastoma classificaton，IIRC) CDE 期、低分化、视神经浸润和淋巴结转移的Rb 患儿血清中NEAT1 表达量明显高于AB 期、中高分化、未发生视神经浸润和淋巴结转移的Rb 患儿，差异具有统计学意义（t=2.190 ～ 3.693，均P<0.05）；血清中NEAT1 表达诊断Rb 曲线下面积为0.882（95%CI：0.826 ～ 0.937），当NEAT1 表达量取1.20 时，灵敏度和特异度分别为80.00% 和79.52%；相比于si-NC 组（1.03 ± 0.09）和Ctl 组（1.02 ± 0.15），si-NEAT1 组细胞中NEAT1 表达（0.35±0.06）明显降低，差异具有统计学意义（t=14.829，9.994，均P<0.001）；si-NEAT1 组24，48，72 和96 h 时吸光度（A 值）明显低于si-NC 组和Ctl 组（tsi-NC=2.796~4.362，tCtl=2.641~5.555，均P<0.05），而细胞凋亡率相比于si-NC 组和Ctl 组明显升高，差异具有统计学意义（t=4.999，3.915，均P<0.05）；与si-NC 组和Ctl 组比较，si-NEAT1 组迁移细胞数（116.50 ± 9.35 vs 132.00 ± 7.32，134.00 ± 7.95）和侵袭细胞数（96.33 ± 8.94 vs 117.67 ± 12.39，119.17 ± 10.05）均降低，差异具有统计学意义（tsi-NC=3.196，3.421，tCtl=3.492，4.159，均P<0.05）。结论 Rb 患儿血清中NEAT1 表达量升高，对Rb 患儿具有一定的诊断价值，沉默Y79细胞中NEAT1 表达可减少Rb 细胞增殖、加速细胞凋亡，同时抑制细胞迁移和侵袭。
Abstract:
Objective To investigate the expression of long non-coding RNA（LncRNA）nuclera-enriched autosomal transcript(NEAT1) in children with retinoblastoma (Rb) and the effect of down-regulation of NEAT1 in Rb cell Y79 on cell biological function. Methods A total of 83 children with Rb who were diagnosed and treated in Huangshi Central Hospital from March 2015 to March 2021 were collected as the research object. During the same period, 50 healthy children (control group) were selected in the children’s health center. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of NEAT1 in serum. The differences in the expressions of NEAT1 in serum between Rb children and the control group, and the differences in the expressions of NEAT1 in serum among Rb children with different clinical indicators, were analyzed. Y79 cells were cultured and were divided into si-NEAT1 group (transfected with interference sequence of NEAT1), si-NC group (transfected with control sequence) and Ctl group (only add transfection reagent). The qRTPCR, MTT, flow cytometry and Transwell were used to detect the NEAT1 expression, cell proliferation, apoptosis, migration and invasion. Results The expression level of NEAT1 in the serum of children with Rb (1.43 ± 0.28) was higher than that in the control group (1.01 ± 0.21), with significant difference (t=9.116, P<0.001). The expression levels of NEAT1 in serum of children with Rb with Intraocular International Retinoblastoma classification (IIRC) stage CDE, poor differentiation, optic nerve infiltration and lymph node metastasis were higher than those in children with Rb with AB, medium to high differentiation, no optic nerve infiltration and lymph node metastasis, with significant differences (t=2.190 ～ 3.693, all P<0.05). The area under the curve for diagnosing Rb based on NEAT1 expression in serum was 0.882 (95%CI: 0.826 ～ 0.937). When the expression level of NEAT1 was 1.20, the sensitivity and specificity were 80.00% and 79.52%, respectively. Compared with the si-NC group (1.03 ± 0.09) and the Ctl group (1.02 ± 0.15), the expression level of NEAT1 in the si-NEAT1 group (0.35 ± 0.06) was decreased, with significant differences (t=14.829, 9.994, all P<0.001). The absorbance A values in the si-NEAT1 group at 24, 48, 72 and 96h were significantly lower than those in the si-NC group and the Ctl group (t=si-NC=2.796~4.362, tCtl=2.641~5.555, all P<0.05), while the apoptosis rate in the si-NEAT1 group was significantly higher than those in the si-NC group and the Ctl group, and the differences were statistically significant (t=4.999, 3.915, all P<0.05). Compared with the si-NC group and the Ctl group, the number of migrating cells (116.50 ± 9.35 vs 132.00 ± 7.32，134.00 ± 7.95) and the number of invasive cells (96.33 ± 8.94 vs 117.67 ± 12.39，119.17 ± 10.05) in the si-NEAT1 group were reduced,and the differences were statistically significant (tsi-NC=3.196, 3.421, tCtl=3.492, 4.159, all P<0.05). Conclusion The expression level of NEAT1 in the serum of children with Rb was elevated, which may have a certain diagnostic value for children with Rb. Silencing the expression of NEAT1 in Y79 cells could reduce cell proliferation, accelerate cell apoptosis, and inhibit cell migration and invasion.

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备注/Memo

备注/Memo:
作者简介：罗文（1984-），女，硕士，主治医师，主要研究方向为眼底病，E-mail：1228755514@qq.com。
通讯作者：张青林（1992-），男，硕士，主治医师，主要研究方向：眼底病，E-mail：827154073@qq.com。
更新日期/Last Update: 2024-05-15