[1]郭宏果a,吴 楠b,陆婉玲a,等.LncRNA01004通过上调CPSF1蛋白表达促进上皮-间质转化加速乳腺癌细胞恶性进展的研究[J].现代检验医学杂志,2025,40(01):32-37+47.[doi:10.3969/j.issn.1671-7414.2025.01.06]
　GUO Hongguoa,WU Nanb,LU Wanlinga,et al.Study on LncRNA01004 Promoting Epithelial-mesenchymal Transformation and Accelerating Malignant Progression of Breast Cancer Cells through Up-regulation of CPSF1 Protein Expression[J].Journal of Modern Laboratory Medicine,2025,40(01):32-37+47.[doi:10.3969/j.issn.1671-7414.2025.01.06]
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LncRNA01004通过上调CPSF1蛋白表达促进上皮-间质转化加速乳腺癌细胞恶性进展的研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年01期
页码:
32-37+47
栏目:
论著
出版日期:
2025-01-15

文章信息/Info

Title:
Study on LncRNA01004 Promoting Epithelial-mesenchymal Transformation and Accelerating Malignant Progression of Breast Cancer Cells through Up-regulation of CPSF1 Protein Expression
文章编号:
1671-7414（2025）01-032-07
作者:
郭宏果a; 吴 楠b; 陆婉玲a; 刘 军a; 程 才a
（中国人民解放军空军第九八六医院 a. 肿瘤血液科；b. 普通外科，西安710054）
Author(s):
GUO Hongguoa;  WU Nanb;  LU Wanlinga;  LIU Juna;  CHENG Caia
(a. Department of Oncology and Hematology；b. Department of General Surgery，the 986th Hospital of the Air Force of the Chinese People’s Liberation Army， Xi’an 710054，China)
关键词:
乳腺癌; 长链非编码RNA 01004; 剪切多聚腺苷酸化特异性因子1; 上皮- 间质转化; 增殖; 侵袭; 凋亡
分类号:
R737.9；R730.43
DOI:
10.3969/j.issn.1671-7414.2025.01.06
文献标志码:
A
摘要:
目的 研究长链非编码RNA 01004（LncRNA01004）加速乳腺癌细胞恶性进展的作用及潜在调节机制。方法 通过实时定量聚合酶链反应（qRT-PCR）检测乳腺癌组织和细胞中LncRNA01004 表达水平。通过StarBase 在线数据库预测LncRNA01004 的结合蛋白，通过RNA 结合蛋白免疫沉淀反应（RIP）分析进行验证。采用LncRNA01004 干扰序列（sh-LncRNA01004）或过表达载体（LncRNA01004）转染MCF-7细胞，或与剪切多聚腺苷酸化特异性因子1（CPSF1）干扰序列（sh-CPSF1）共转染MCF-7 细胞。CCK-8 法、Transwell 和流式细胞术（FCM）分别检测细胞活力、细胞侵袭和凋亡。qRT-PCR 检测LncRNA01004 和CPSF1 过表达和沉默效率；蛋白质印迹（WB）检测CPSF1 蛋白、凋亡相关蛋白[B 淋巴细胞瘤-2（Bcl-2），Bcl-2 相关X蛋白（Bax）]，以及上皮- 间质转化（EMT）相关蛋白[ 上皮钙黏蛋白（E-cadherin）、神经钙黏蛋白（N-cadherin）、波形蛋白（Vimentin）、锌指转录因子（Snail）] 水平；ELISA 检测半胱天冬氨酸蛋白酶3（Caspase-3）活性。结果 与癌旁组织相比，LncRNA01004 在乳腺癌组织（5.14±0.33 vs 1.02±0.03）中显著上调，差异具有统计学意义（t=-78.637，P<0.001）；乳腺癌细胞中LncRNA01004 表达亦明显高于正常人乳腺上皮细胞，组间差异具有统计学意义（F=142.248，P<0.001）。与Control 组相比，沉默LncRNA01004 显著抑制MCF-7 细胞活力（42.15±2.11 vs 100.02±0.65）和侵袭（18.65%±1.44% vs 41.36%±1.57%），诱导细胞凋亡（16.58%±1.52% vs5.24%±1.12%），升高Caspase-3 活性（2.93±0.71 vs 1.51±0.43）和Bax 蛋白（2.74±0.39 vs 1.01±0.02）表达，抑制BcL-2 蛋白（0.32±0.07 vs 1.02±0.03）表达，差异具有统计学意义（t=3.075 ～ 19.332，均P<0.05）。与Control 组相比，沉默LncRNA01004 显著升高E-cadherin（3.06±0.37 vs 1.01±0.02）蛋白水平，降低N-cadherin（0.44±0.11 vs 1.00±0.01），Vimentin（0.39±0.13 vs 1.02±0.03）和Snail（0.30±0.08 vs 1.01±0.03）蛋白水平，差异具有统计学意义（t=9.989 ～ 17.164，均P<0.05）。LncRNA01004 与CPSF1 结合并促进CPSF1 蛋白表达。沉默CPSF1 抑制MCF-7 细胞增殖和侵袭、诱导细胞凋亡，抵消LncRNA01004 过表达对MCF-7 细胞生物学功能的影响。结论 LncRNA01004 可能通过上调CPSF1 促进EMT，进而促进乳腺癌细胞增殖和侵袭，抑制细胞凋亡，参与乳腺癌的恶性进展。
Abstract:
Objective To investigate the role of long non-coding RNA 01004 (LncRNA01004) in accelerating the malignant progression of breast cancer cells and its potential regulatory mechanism. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression level of LncRNA01004 in breast cancer tissues and cells. The Starbase online database predicted the binding of LncRNA01004 to CPSF1 and verified it through RNA binding protein immunoprecipitation(RIP) analysis. MCF-7 cells were transfected with LncRNA01004 interference sequence (sh-LncRNA01004) or overexpressed vector (LncRNA01004), or co-transfected with Cleavage and Polyadenylation Specific Factor 1 (CPSF1) interference sequence (sh-CPSF1). Cell viability, invasion and apoptosis were detected with CCK-8, Transwell and flow cytometry (FCM). The overexpression and silencing efficiency of LINC01004 and CPSF1 were detected by qPCR. Western blot (WB) analysis of CPSF1 protein, apoptosis-related protein [B cell lymphoma/leukemia-2(Bcl-2), Bcl-2 Associated X(Bax)] and Epithelial-Mesenchymal transition (EMT) related protein[Epitheia-cadherin(E-cadherin), Nerve-cadherin(N-cadherin), Vimentin, zinc-finger transcription factor(Snail)], and the activity of Cysteinyl aspartate-specific proteinase-3 (Caspase-3) was determined by enzyme-linked immunosorbent assay. Results LncRNA01004 was significantly up-regulated in breast cancer tissues (5.14±0.33) compared with paracancer tissues (1.02±0.03), with the statistically significant difference (t=-78.637，P<0.001); LncRNA01004 expression in breast cancer cells was significantly higher than that in normal breast epithelial cells， the difference between groups is statistically significant (F=142.248，P<0.001). Compared with the Control group, LncRNA01004 significantly inhibited the proliferation (42.15±2.11 vs 100.02±0.65) and invasion (18.65%±1.44% vs 41.36%±1.57%) of MCF-7 cells, induced apoptosis (16.58%±1.52% vs 5.24%±1.12%), increased the activity of Caspase-3 (2.93±0.711. vs 51±0.43) and the expression of Bax (2.74±0.39 vs 1.01±0.02) protein, and inhibited the expression of BcL-2 (0.32±0.07 vs 1.02±0.03) protein, with the statistically significant difference (t=3.075 ～ 19.332，all P<0.05). Significantly increased compared with Control group, the silent LncRNA01004 E-cadherin (3.06±0.37 vs 1.01±0.02) protein levels, lower N-cadherin (0.44±0.11 vs 1.00±0.01), Vimentin (0.39±0.13 vs 1.02±0.03) and Snail(0.30±0.08 vs 1.01±0.03) protein levels, with the statistically significant difference (t=9.989 ～ 17.164，all P<0.05).LncRNA01004 binds to CPSF1 and promotes CPSF1 protein expression.Silencing CPSF1 inhibited the proliferation and invasion of MCF-7 cells, induced cell apoptosis, and counteracted the effect of LncRNA01004 overexpression on MCF-7 cells. Conclusion LncRNA01004 may promote EMT through up-regulation of CPSF1, and then promote proliferation and invasion of breast cancer cells, inhibit cell apoptosis, and participate in the malignant progression of breast cancer.

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备注/Memo

备注/Memo:
作者简介：郭宏果（1979-），女，硕士研究生，主治医师，研究方向：乳腺癌、妇科肿瘤，E-mail：guohongguo_986@163.com.
通讯作者：吴楠（1991-），女，硕士研究生，主治医师，研究方向：乳腺癌、甲状腺癌治疗，E-mail：617302210@qq.com.
更新日期/Last Update: 2025-01-15