PDF(Pubmed)
Abstract：
Lung adenocarcinoma (LUAD) is the leading cause of death among cancer diseases. The tumorigenic functions of AHNAK2 in LUAD have attracted more attention in recent years, while there are few studies which have reported its high molecular weight.
The mRNA-seq data of AHNAK2 and corresponding clinical data from UCSC Xena and GEO was analyzed. LUAD cell lines were transfected with sh-NC and sh-AHNAK2, and cell proliferation, migration and invasion were then detected by in vitro experiments. We performed RNA sequencing and mass spectrometry analysis to explore the downstream mechanism and interacting proteins of AHNAK2. Finally, western blot, cell cycle analysis and CO-IP were used to confirm our assumptions regarding previous experiments.
Our study revealed that AHNAK2 expression was significantly higher in tumors than in normal lung tissues and higher AHNAK2 expression led to a poor prognosis, especially in patients with advanced tumors. AHNAK2 suppression via shRNA reduced the LUAD cell lines proliferation, migration and invasion and induced significant changes in DNA replication, NF-kappa B signaling pathway and cell cycle. AHNAK2 knockdown also caused G1/S phase cell cycle arrest, which could be attributed to the interaction of AHNAK2 and RUVBL1. In addition, the results from gene set enrichment analysis (GSEA) and RNA sequencing suggested that AHNAK2 probably plays a part in the mitotic cell cycle.
AHNAK2 promotes proliferation, migration and invasion in LUAD and regulates the cell cycle via the interaction with RUVBL1. More studies of AHNAK2 are still needed to reveal its upstream mechanism.
The mRNA-seq data of AHNAK2 and corresponding clinical data from UCSC Xena and GEO was analyzed. LUAD cell lines were transfected with sh-NC and sh-AHNAK2, and cell proliferation, migration and invasion were then detected by in vitro experiments. We performed RNA sequencing and mass spectrometry analysis to explore the downstream mechanism and interacting proteins of AHNAK2. Finally, western blot, cell cycle analysis and CO-IP were used to confirm our assumptions regarding previous experiments.
Our study revealed that AHNAK2 expression was significantly higher in tumors than in normal lung tissues and higher AHNAK2 expression led to a poor prognosis, especially in patients with advanced tumors. AHNAK2 suppression via shRNA reduced the LUAD cell lines proliferation, migration and invasion and induced significant changes in DNA replication, NF-kappa B signaling pathway and cell cycle. AHNAK2 knockdown also caused G1/S phase cell cycle arrest, which could be attributed to the interaction of AHNAK2 and RUVBL1. In addition, the results from gene set enrichment analysis (GSEA) and RNA sequencing suggested that AHNAK2 probably plays a part in the mitotic cell cycle.
AHNAK2 promotes proliferation, migration and invasion in LUAD and regulates the cell cycle via the interaction with RUVBL1. More studies of AHNAK2 are still needed to reveal its upstream mechanism.
摘要：
背景：肺腺癌（LUAD）是导致癌症死亡的主要原因。近年来，AHNAK2在LUAD中的致瘤功能引起了越来越多的关注，虽然很少有研究报道其高分子量。
方法：分析了AHNAK2的mRNA-seq数据以及来自UCSCXena和GEO的相应临床数据。用sh-NC和sh-AHNAK2转染LUAD细胞系,然后通过体外实验检测迁移和侵袭。我们进行了RNA测序和质谱分析，以探索AHNAK2的下游机制和相互作用蛋白。最后,westernblot,使用细胞周期分析和CO-IP来证实我们对先前实验的假设.
结果：我们的研究表明，AHNAK2在肿瘤中的表达明显高于正常肺组织，而较高的AHNAK2表达导致预后不良，尤其是晚期肿瘤患者。通过shRNA抑制AHNAK2降低了LUAD细胞系的增殖，迁移和入侵，并诱导DNA复制的显着变化，NF-κB信号通路与细胞周期.AHNAK2敲低也引起G1/S期细胞周期停滞，这可能归因于AHNAK2和RUVBL1的相互作用。此外,基因集富集分析（GSEA）和RNA测序的结果表明，AHNAK2可能在有丝分裂细胞周期中起作用。
结论：AHNAK2促进增殖，LUAD中的迁移和侵袭，并通过与RUVBL1的相互作用调节细胞周期。AHNAK2还需要更多的研究来揭示其上游机制。
方法：分析了AHNAK2的mRNA-seq数据以及来自UCSCXena和GEO的相应临床数据。用sh-NC和sh-AHNAK2转染LUAD细胞系,然后通过体外实验检测迁移和侵袭。我们进行了RNA测序和质谱分析，以探索AHNAK2的下游机制和相互作用蛋白。最后,westernblot,使用细胞周期分析和CO-IP来证实我们对先前实验的假设.
结果：我们的研究表明，AHNAK2在肿瘤中的表达明显高于正常肺组织，而较高的AHNAK2表达导致预后不良，尤其是晚期肿瘤患者。通过shRNA抑制AHNAK2降低了LUAD细胞系的增殖，迁移和入侵，并诱导DNA复制的显着变化，NF-κB信号通路与细胞周期.AHNAK2敲低也引起G1/S期细胞周期停滞，这可能归因于AHNAK2和RUVBL1的相互作用。此外,基因集富集分析（GSEA）和RNA测序的结果表明，AHNAK2可能在有丝分裂细胞周期中起作用。
结论：AHNAK2促进增殖，LUAD中的迁移和侵袭，并通过与RUVBL1的相互作用调节细胞周期。AHNAK2还需要更多的研究来揭示其上游机制。
