[1]沈志勇,王 玲,朱正安,等.基于TCGA数据库生物学信息分析FEN1与肺腺癌相关性并敲低FEN1基因抑制小鼠Lewis肺癌细胞生长及其分子机制的实验研究[J].现代检验医学杂志,2026,41(02):174-180.[doi:10.3969/j.issn.1671-7414.2026.02.029]
　SHEN Zhiyong,WANG Ling,ZHU Zhengan,et al.TCGA-Based Analysis of FEN1 in Lung Adenocarcinoma and the Molecular Mechanism of FEN1 Knockdown Inhibiting the Growth of Lewis Lung Cancer Cells in Mice[J].Journal of Modern Laboratory Medicine,2026,41(02):174-180.[doi:10.3969/j.issn.1671-7414.2026.02.029]
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基于TCGA数据库生物学信息分析FEN1与肺腺癌相关性并敲低FEN1基因抑制小鼠Lewis肺癌细胞生长及其分子机制的实验研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
174-180
栏目:
论著
出版日期:
2026-03-15

文章信息/Info

Title:
TCGA-Based Analysis of FEN1 in Lung Adenocarcinoma and the Molecular Mechanism of FEN1 Knockdown Inhibiting the Growth of Lewis Lung Cancer Cells in Mice
文章编号:
1671-7414（2026）02-174-07
作者:
沈志勇1,2，王 玲1,2，朱正安1,2，林秀华3，傅志超1,2，陈忠华1,2
1.福建医科大学福总临床医学院放射治疗科，福州 350025;2.中国人民解放军联勤保障部队第九〇〇医院放射治疗科，福州 350025;3.福建医科大学孟超肝胆医院呼吸内科，福州350028
Author(s):
SHEN Zhiyong1,2，WANG Ling1,2，ZHU Zhengan1,2，LIN Xiuhua3，FU Zhichao1,2，CHEN Zhonghua1,2
1.Department of Radiotherapy, Fuzong School of Clinical Medicine College of Fujian Medical University, Fuzhou 350025, China;2.Department of Radiotherapy, 900th Hospital of PLA Joint Logistic Support Force, Fuzhou 350025, China;3.Respiratory Medicine, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou 350028, China
关键词:
瓣状核酸内切酶1; 肺腺癌; 路易斯肺癌细胞; 上皮-间充质转化; 细胞凋亡
分类号:
R-332
DOI:
10.3969/j.issn.1671-7414.2026.02.029
文献标志码:
A
摘要:
目的?基于癌症基因组图谱(TCGA)数据库分析瓣状核酸内切酶1(FEN1)在肺腺癌(LUAD)中的表达及预后价值,并阐明其调控LUAD细胞生物学行为的分子机制。方法分析TCGA数据库598例样本(539例LUAD组织、59例正常组织)转录组数据,评估FEN1表达与患者预后关系。构建FEN1敲低的小鼠Lewis肺癌细胞(LLC),按转染情况分为LLC组、shRNA-NC组和FEN1-shRNA组。采用细胞计数试剂盒-8(CCK8)法、流式细胞术(FCM)、Transwell法分别检测细胞增殖、凋亡、周期及迁移、侵袭能力。实时荧光定量PCR(RT-qPCR)和蛋白印迹(Westernblot)检测上皮-间充质转化(EMT)标志物[上皮型钙黏蛋白(E-cadherin)、神经型钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)]及细胞周期蛋白(Cyclin)D1、B细胞淋巴瘤/白血病-2基因(Bcl-2)、基质金属蛋白酶-9(MMP-9)的表达。结果?FEN1在LUAD组织中高表达,与不良预后显著相关(Log-rank检验,Z=2.81,P=0.005)。成功构建FEN1稳定敲低细胞株,敲低效率83.8%。与LLC组相比,FEN1-shRNA组24、48h存活率降低(62.31%±2.84%vs100%±0.93%、81.58%±2.35%vs100.00%±4.88%),凋亡率增高(25.82%±0.30%vs3.19%±0.46%),差异具有统计学意义(t=30.892、8.336、-70.889,均P<0.001);细胞周期阻滞于S期,S期比例升高(71.07%±3.27%vs47.85%±4.15%),差异具有统计学意义(F=43.688,P<0.001)。迁移细胞数(118.556±12.738vs192.222±14.986)、侵袭细胞数(92.667±3.000vs129.000±9.042)均显著降低,差异具有统计学意义(t=6.487、6.606,均P<0.01)。分子机制显示,FEN1敲低逆转EMT进程:E-cadherin蛋白和mRNA上调(t=-10.478、-4.235),N-cadherin、Vimen-tin、CyclinD1、Bcl-2、MMP-9mRNA和蛋白均下调(t=-20.825~33.396),差异具有统计学意义(均P<0.05)。结论FEN1在LUAD中高表达且与不良预后相关,通过调控EMT通路促进LUAD细胞恶性表型。FEN1敲低抑制细胞增殖、迁移侵袭并促进凋亡,可作为LUAD预后评估的生物学标志物和潜在治疗靶点。
Abstract:
Objective To analyze the expression characteristics and prognostic value of Flap Endonuclease 1(FEN1)in lung ad-enocarcinoma based on The Cancer Genome Atlas (TCGA) database and elucidate the molecular mechanisms by which FEN1 regulates the biological behavior of lung cancer cells. Methods Transcriptomic data from 598 samples (539 lung adenocarcino-ma tissues and 59 normal tissues) in the TCGA database were analyzed to evaluate the relationship between FEN1 expression and patient prognosis. Lewis lung carcinoma (LLC) cells with FEN1 knockdown were established and divided into LLC group, shRNA-NC group, and FEN1-shRNA group based on transfection status. Cell proliferation, apoptosis, cell cycle progression,mi-gration and invasion abilities were detected using CCK-8 assay, flow cytometry and Transwell assay, respectively. The expression of epithelial-mesenchymal transition (EMT) markers (E-cadherin, N-cadherin, Vimentin) and cell cycle protein Cyclin D1, Bcl-2 and MMP9 were measured by RT-qPCR and Western blot. Results FEN1 was highly expressed in lung adenocarcinoma tissues and significantly correlated with poor prognosis (Log-rank test, Z=2.81, P=0.005). A stable FEN1 knockdown cell lines were suc-cessfully constructed with a knockdown efficiency of 83.8%. Compared with LLC groups, the survival rates of FEN1-shRNA group at 24h and 48h decreased [(62.31±2.84)% vs (100.00±0.93)% and [(81.58±2.35)% vs (100.00±4.88)%], while the apoptosis rate increased to [(25.82±0.30)% vs (3.19±0.46)%], and the differences were statistically significant (t=30.892, 8.336,-70.889, all P<0.001). Cell cycle was arrested at S phase with the S phase proportion increased to [(71.07±3.27)% vs (47.85±4.15)%] (F=43.688, P<0.001). The number of migrated cells (118.556±12.738 vs 192.222±14.986) and invaded cells (92.667±3.000 vs 129.000±9.042) were significantly decreased, and the differences were statistically significant (t=6.487, 6.606, all P<0.001). Molecular mechanism analysis revealed that FEN1 knockdown reversed the EMT process: E-cadherin pro-tein and mRNA were upregulated (t=-10.478, -4.235), while N-cadherin, Vimentin, Cyclin D1, Bcl-2, and MMP9 protein and mRNA were all downregulated (t=-20.825~33.396), and the differences were statistically significant (all P<0.05). Conclusions FEN1 is highly expressed in lung adenocarcinoma and associates with poor prognosis. FEN1 promotes the malignant phenotypes of lung cancer cells by regulating the EMT pathway. FEN1 knockdown inhibits cell proliferation, migration and invasion while promoting apoptosis, suggesting that FEN1 can serve as an important biomarker for prognostic assessment and a potential thera-peutic target for lung adenocarcinoma.

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备注/Memo

备注/Memo:
基金项目：福建省自然科学基金项目（2023J011360）。
作者简介：沈志勇（1981-），男，医学硕士，副主任医师，研究方向：肺癌的发病机制及肺癌的精准靶向治疗与放疗，E-mail：shlinxiu123@126.com。
通讯作者：陈忠华（1970-），男，医学硕士，主任医师，研究方向：恶性肿瘤精准放疗，E-mail：zyczhonghua@163.com。
更新日期/Last Update: 2026-03-15