<div>磷酸丙糖异构酶1对不同类型肺癌细胞生物学功能的影响</div>

刘尚书; 李春笋; 王平; 刘鹿; 任佳博; 王梓瑞; 殷悦; 陈良安

doi:10.12435/j.issn.2095-5227.24112705

磷酸丙糖异构酶1对不同类型肺癌细胞生物学功能的影响

Effects of Triosephosphate isomerase 1 on the biological functions of different types of lung cancer cells

摘要

摘要:
背景　磷酸丙糖异构酶1(triosephosphate isomerase 1，TPI1)与多种肿瘤的形成发展密切相关，但在肺癌中的研究尚不充分。目的　探讨TPI1在不同类型肺癌细胞系中的表达水平及生物学功能。方法　应用TCGA数据库，通过生物信息学分析TPI1在肺腺癌和鳞癌组织中的表达；采用定量PCR和蛋白质免疫印迹检测TPI1在不同肺癌细胞系中的表达。采用慢病毒稳定感染技术分别建立肺腺癌细胞A549、肺鳞癌细胞Calu1和肺大细胞癌细胞95D的TPI1敲低细胞株，于mRNA水平验证敲低效率。分别采用CCK-8法、黏附实验、划痕实验、Transwell实验和流式细胞术检测细胞的增殖、黏附、迁移、侵袭能力和细胞周期。应用普通转录组测序技术和KEGG通路富集分析初步探索可能的机制。结果　生物信息学分析显示，与正常肺组织相比，TPI1在肺腺癌和肺鳞癌组织中表达上调(P＜0.05)；相较于肺泡上皮细胞系HPAEpic，肺腺癌细胞系A549、肺鳞癌细胞系Calu1和肺大细胞癌细胞系95D中TPI1在mRNA水平和蛋白水平均呈高表达(均P＜0.05)，肺小细胞癌细胞系H446中TPI1在mRNA水平呈低表达(P＜0.05)，蛋白水平表达无统计学差异(P＞0.05)。在A549、Calu1和95D细胞中敲低TPI1后，细胞增殖、黏附、迁移和侵袭能力均下降，细胞周期发生G0/G1期阻滞(均P＜0.05)。KEGG通路富集分析结果显示，差异基因显著富集在PI3K/AKT信号通路和MAPK信号通路。结论 TPI1在肺腺癌A549、肺鳞癌Calu1和肺大细胞癌95D细胞系中高表达，可能通过PI3K/AKT和MAPK信号通路调控细胞增殖、黏附、迁移、侵袭及周期，推动肺癌的发展。

Abstract:
Background Triosephosphate isomerase 1 (TPI1) is closely associated with the development of various tumors, but its role in lung cancer has not been fully investigated.Objective To explore the expression levels and biological functions of TPI1 in different types of lung cancer cell lines. Methods The expression of TPI1 in lung adenocarcinoma and squamous carcinoma tissues was analyzed using bioinformatics from the TCGA database. Quantitative PCR and Western blotting were employed to detect TPI1 expression in various lung cancer cell lines. Lentivirus-mediated stable infection technology was used to establish TPI1 knockdown cell lines in lung adenocarcinoma A549 cells, squamous carcinoma Calu1 cells, and large cell lung cancer 95D cells. The knockdown efficiency was verified at the mRNA level. Cell proliferation, adhesion, migration, invasion, and cell cycle were assessed by CCK-8 assay, adhesion assay, scratch assay, Transwell assay, and flow cytometry, respectively. RNA sequencing and KEGG pathway enrichment analysis were applied to explore potential mechanisms. Results Bioinformatics analysis showed that, compared to normal lung tissues, TPI1 expression was upregulated in lung adenocarcinoma and squamous carcinoma tissues (P<0.05). Compared to the alveolar epithelial cell line HPAEpic, TPI1 was highly expressed at both the mRNA
and protein levels in lung adenocarcinoma A549, squamous carcinoma Calu1, and large cell lung cancer 95D cell lines (all P<0.05), while TPI1 expression was low at the mRNA level in the small cell lung cancer H446 cell line (P<0.05), with no significant difference at the protein level (P>0.05). After TPI1 knockdown in A549, Calu1, and 95D cells, cell proliferation, adhesion, migration, and invasion abilities were significantly decreased, and cell cycle arrest occurred in the G0/G1 phase (all P<0.05). KEGG pathway enrichment analysis revealed that differentially expressed genes were significantly enriched in the PI3K/AKT and MAPK signaling pathways.Conclusion TPI1 is highly expressed in lung adenocarcinoma A549, squamous carcinoma Calu1, and large cell lung cancer 95D cell lines, and may regulate cell proliferation, adhesion, migration, invasion, and cell cycle through the PI3K/AKT and MAPK signaling pathways, promoting lung cancer development.

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