LncRNA PKD2-2-3对肺腺癌细胞增殖、克隆形成、迁移及侵袭能力的影响

doi:10.19401/j.cnki.1007-3639.2023.08.001

中国癌症杂志 ›› 2023, Vol. 33 ›› Issue (8): 717-725.doi: 10.19401/j.cnki.1007-3639.2023.08.001

LncRNA PKD2-2-3对肺腺癌细胞增殖、克隆形成、迁移及侵袭能力的影响

贾利晴¹(), 葛小路², 姜琳¹, 周晓燕¹()

1.复旦大学附属肿瘤医院病理科，复旦大学上海医学院肿瘤学系，复旦大学病理研究所，上海 200032
2.上海市第一人民医院疑难疾病精准研究中心，上海 201620

收稿日期:2023-05-01 修回日期:2023-07-05 出版日期:2023-08-30 发布日期:2023-09-01
通信作者: 周晓燕（ORCID：0000-0001-5999-7237），主任医师，教授。
作者简介:贾利晴（ORCID：0000-0003-1734-6043），硕士。
基金资助:
国家自然科学基金(82002424)

Effects of lncRNA PKD2-2-3 on cell proliferation, clone formation, migration, and invasion of lung adenocarcinoma

JIA Liqing¹(), GE Xiaolu², JIANG Lin¹, ZHOU Xiaoyan¹()

1. Department of Pathology, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College, Fudan University; Fudan University Cancer Institute, Shanghai 200032, China
2. Precision Research Center for Refractory Diseases, Shanghai General Hospital, Shanghai 201620, China

Received:2023-05-01 Revised:2023-07-05 Published:2023-08-30 Online:2023-09-01
Contact: ZHOU Xiaoyan

摘要/Abstract

摘要：

背景与目的：长链非编码RNA（long non-coding RNA，lncRNA）在肺腺癌患者中异常表达，与肿瘤的发生、发展和化疗耐药密切相关。本研究旨在探讨lncRNA蛋白激酶D2（lncRNA PKD2-2-3）在肺腺癌发生、发展过程中发挥的生物学功能，验证lncRNA PKD2-2-3对肺腺癌细胞增殖、克隆形成、迁移和侵袭的影响。方法：基于Affymetrix人类基因芯片转录组阵列2（Affymetrix® GeneChip Human Transcriptome Array 2.0，HTA2.0）分析3对肺腺癌组织及癌旁组织，寻找在肺腺癌组织中高表达的lncRNA，其中lncRNA PKD2-2-3的表达差异最大。利用基因表达综合数据库（Gene Expression Omnibus，GEO）中GSE19188和GSE30219的数据分析lncRNA PKD2-2-3在肺腺癌组织中的表达和对患者预后的影响。通过实时荧光定量聚合酶链反应（real-time fluorescence quantitative polymerase chain reaction，RTFQ-PCR）检测lncRNA PKD2-2-3在细胞系（HBE、A549、PC9）中的表达情况。在A549和PC9两种细胞系中使用siRNA干扰技术敲低lncRNA PKD2-2-3的表达，通过细胞计数试剂盒-8（cell counting kit-8，CCK-8）实验和克隆形成实验检测lncRNA PKD2-2-3表达对肺腺癌细胞增殖和克隆形成能力的影响；划痕实验和transwell实验分别检测lncRNA PKD2-2-3表达对肺腺癌细胞迁移和侵袭的影响。蛋白质印迹法（Western blot）检测敲低lncRNA PKD2-2-3表达后对上皮-间质转化（epithelial-mesenchymal transition，EMT）相关蛋白E-钙黏蛋白（E-cadherin）和N-钙黏蛋白（N-cadherin）表达水平的影响。皮下移植瘤模型探究lncRNA PKD2-2-3在体内对肺腺癌细胞增殖的影响。结果：LncRNA PKD2-2-3在肺腺癌组织中呈高表达，且高表达的患者预后较差。对比正常气管上皮永生化细胞HBE，lncRNA PKD2-2-3在肺腺癌细胞系A549和PC9中高表达，敲低lncRNA PKD2-2-3的表达抑制肺腺癌细胞的增殖、克隆形成、迁移和侵袭能力。Western blot结果显示，敲低lncRNA PKD2-2-3后，E-cadherin表达水平升高，N-cadherin表达水平降低。皮下移植瘤实验表明，在体内lncRNA PKD2-2-3敲低后抑制肺腺癌的生长。结论：ncRNA PKD2-2-3在肺腺癌组织中上调表达，且与患者的不良预后相关，lncRNA PKD2-2-3的高表达促进肺腺癌细胞的增殖、克隆形成、迁移和侵袭能力，体内外实验表明lncRNA PKD2-2-3的表达与肺腺癌的EMT过程密切相关。

关键词: 肺腺癌, lncRNA PKD2-2-3, 细胞迁移, 细胞侵袭, 上皮间质转化

Abstract:

Background and purpose: Long non-coding RNA (lncRNA) is abnormally expressed in lung adenocarcinoma patients, and closely related to tumor occurrence, development and chemotherapy resistance. In this study, we mainly investigated the biological function of lncRNA PKD2-2-3 and verified its effect on the proliferation, colony formation, migration and invasion in lung adenocarcinoma. Methods: Three pairs of lung adenocarcinoma tissues and adjacent tissues were analyzed based on expression profiling Affymetrix® GeneChip Human Transcriptome Array 2.0 (HTA2.0), and we focused on lncRNA PKD2-2-3 that showed most significant difference between lung adenocarcinoma issues and adjacent tissues. Besides, we found the upregulated expression of lncRNA PKD2-2-3 in lung adenocarcinoma tissues and suggested the relation of lncRNA PKD2-2-3 expression with prognosis by using GSE19188 and GSE30219 data in Gene Expression Omnibus (GEO) database. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of lncRNA PKD2-2-3 in cell lines including HBE, A549 and PC9. After using siRNAs to decrease the expression of lncRNA PKD2-2-3 in A549 and PC9, we detected cell proliferation and colony formation by cell counting kit-8 (CCK-8) assay and colony formation assay. Effects of lncRNA PKD2-2-3 on migration and invasion in lung adenocarcinoma cells were detected by wound-healing assay and transwell assay, respectively. Moreover, we detected expression levels of E-cadherin and N-cadherin that were epithelial-mesenchymal transition (EMT) related genes by Western blot. The effect of lncRNA PKD2-2-3 on the formation and growth of lung adenocarcinoma in vivo was verified by subcutaneous transplantation tumor model. Results: LncRNA PKD2-2-3 was highly expressed in lung adenocarcinoma tissue, and was positively associated with poor prognosis of lung adenocarcinoma patients. Compared with human bronchial epithelial cells (HBE), lncRNA PKD2-2-3 was overexpressed in A549 and PC9. The proliferation, colony formation, migration and invasion of lung adenocarcinoma cells were significantly inhibited when decreasing the expression level of lncRNA PKD2-2-3. Western blot also showed that the expression level of E-cadherin was increased, while the level of N-cadherin was decreased after lncRNA PKD2-2-3 knockdown. Subcutaneous tumor transplantation experiments showed that lncRNA PKD2-2-3 knockdown inhibited the growth of lung adenocarcinoma in vivo. Conclusion: LncRNA PKD2-2-3 expression was upregulated in lung adenocarcinoma tissues, and it was associated with poor prognosis of lung adenocarcinoma patients. Overexpression of lncRNA PKD2-2-3 promoted the proliferation, colony formation, migration and invasion of lung adenocarcinoma cells. LncRNA PKD2-2-3 level was closely related to EMT process in lung adenocarcinoma in vitro and in vivo.

Key words: Lung adenocarcinoma, LncRNA PKD2-2-3, Cell migration, Cell invasion, Epithelial-mesenchymal transition

中图分类号:

R734.2

贾利晴, 葛小路, 姜琳, 周晓燕. LncRNA PKD2-2-3对肺腺癌细胞增殖、克隆形成、迁移及侵袭能力的影响[J]. 中国癌症杂志, 2023, 33(8): 717-725.

JIA Liqing, GE Xiaolu, JIANG Lin, ZHOU Xiaoyan. Effects of lncRNA PKD2-2-3 on cell proliferation, clone formation, migration, and invasion of lung adenocarcinoma[J]. China Oncology, 2023, 33(8): 717-725.

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参考文献 16

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LncRNA PKD2-2-3对肺腺癌细胞增殖、克隆形成、迁移及侵袭能力的影响

Effects of lncRNA PKD2-2-3 on cell proliferation, clone formation, migration, and invasion of lung adenocarcinoma

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