SPC에 의해 유도되는 상피 간엽 천이과정에 미치는 Ethacrynic acid의 억제 효과에 관한 연구 = Ethacrynic acid inhibits Sphingosylphosphorylcholine (SPC)-induced epithelial-mesenchymal transition in A549 lung cancer cells

다국어 초록 (Multilingual Abstract)

Lung cancer is a fatal disease that accounts for 14% of new cancer cases. Metastasis is the primary cause of death in patients with lung cancer. Epithelial-mesenchymal transition (EMT) contributes to lung cancer invasion and metastasis. Therefore, it is crucial to find appropriate therapy for lung cancer and EMT.
Ethacrynic acid (ECA) is a diuretic that inhibits cellular ion flux that leads to an increase in intracellular Na concentrations.
We studied the effects of ethacrynic acid (ECA) on sphingosylphosphorylcholine (SPC)-induced EMT in A549 lung cancer cells by RT-PCR, Western blot, and RNA sequencing.
We found that ECA inhibits SPC-induced EMT in A549 lung cancer cells. ECA inhibited SPC-induced migration and invasion in A549 lung cancer cells. We identified norrin (NDP) and Wnt2 as ECA responsive genes in A549 lung cancer cells through transcriptome analysis by RNA seq. NDP and WNT2 were validated by RT-PCR. NDP is found to be involved in SPC-induced WNT activation by NDP si-RNA.
These results suggested that ECA suppresses SPC-induced EMT of A549 lung cancer cells via downregulation of NDP expression and ECA might be used as an anti-metastatic drug for lung cancer.

목차 (Table of Contents)

• Abstract i
• Table of Contents iii
• List of Figures v
• List of Abbreviations vii
• Graphic Abstract viii
• Introduction 1
• Materials and methods 8
• Materials 8
• Cell culture 8
• Western blot 8
• RT-PCR 9
• Confocal microscopy 10
• Invasion assay 10
• Migration assay 11
• Wound healing assay 11
• RNA-seq 11
• Transfections (siRNA) 12
• Statistical analysis 12
• Results 13
• ECA suppresses SPC-induced EMT process of A549 lungcancer cells 13
• ECA suppresses SPC-induced the migration of A549 lungcancer cells 20
• ECA suppresses SPC-induced the invasion of A549 lungcancer cells 23
• ECA suppresses TGF-β-induced EMT process and migration and invasion in A549 lung cancer cells 25
• Identification of ECA responsive genes in A549 lung cancer cells through transcriptome analysis by RNA seq 34
• ECA suppresses SPC-induced the activation of Wnt/beta-catenin signaling in A549 lung cancer cells 38
• NDP is involved in SPC-induced events of lung cancer cells 40
• Discussion 46
• References 48