DDP Induced Cytotoxicity through miR-215-5p/COL5A1/FSTL1 Axis to Regulate Autophagy in Lung Adenocarcinoma Cells
Xinguo Zhao1, Hongming Zhang1, Longqiang Gu1
1Department of Pneumology, Affiliated Hospital 6 of Nantong University, The Yancheng School of Clinical Medicine of Nanjing Medical University, Yancheng Third People's Hospital, 224000, Yancheng, China.
Background:
Cisplatin (DDP) resistance remains a major challenge in the treatment of lung adenocarcinoma (LUAD). Autophagy is an important mechanism to generate drug resistance. It has been established that COL5A1 has been shown to accelerate LUAD metastasis and affect cellular processes.
Methods:
We investigated the role of COL5A1 in DDP resistance using the H1299/DDP and A549/DDP cell lines. Flow cytometry, CCK8, and western blot assays were used to detect apoptosis, cell viability, and autophagy. In addition, upstream miRNAs were screened using bioinformatics methods. MS2-RIP assay and luciferase reporter gene assay were used to validate miRNA interaction with COL5A1. Transfection experiments and western blot experiments were performed to investigate miRNA targeting to COL5A1 and its regulation of autophagy through FSTL1. The role of miRNA and COL5A1 in LUAD cisplatin resistance was also verified in vivo.
Results:
The down-regulation of COL5A1 significantly reduced the survival and autophagy of DDP-resistant cells while enhancing apoptosis. MiR-215-5p was found to be a direct regulator of COL5A1, which affects autophagy through FSTL1.
Conclusion:
The present study demonstrated that miR-215-5p regulated COL5A1 to modulate FSTL1 and autophagy, thereby attenuating LUAD resistance to DDP. These findings deepen the understanding of LUAD pathogenesis and provide potential insights into therapeutic strategies.
Insights
MicroRNA-215-5p targets COL5A1 to reduce autophagy and overcome cisplatin resistance in lung adenocarcinoma. This finding offers new therapeutic strategies for treating drug-resistant lung cancer.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Cisplatin (DDP) resistance is a significant obstacle in treating lung adenocarcinoma (LUAD).
- Autophagy plays a crucial role in developing drug resistance.
- COL5A1 is implicated in promoting LUAD metastasis and influencing cellular functions.
Purpose of the Study:
- To investigate the role of COL5A1 in DDP resistance in LUAD.
- To identify upstream regulators of COL5A1 involved in DDP resistance.
- To elucidate the molecular mechanisms by which COL5A1 affects DDP resistance and autophagy.
Main Methods:
- Utilized DDP-resistant LUAD cell lines (H1299/DDP, A549/DDP).
- Assessed apoptosis, cell viability, and autophagy via flow cytometry, CCK8, and western blot.
- Employed bioinformatics, MS2-RIP, and luciferase assays to identify and validate miRNA-COL5A1 interactions.
- Conducted in vitro and in vivo experiments to confirm the regulatory pathway involving miR-215-5p, COL5A1, FSTL1, and autophagy.
Main Results:
- Down-regulation of COL5A1 decreased survival and autophagy while increasing apoptosis in DDP-resistant LUAD cells.
- MiR-215-5p was identified as a direct regulator of COL5A1.
- MiR-215-5p modulates autophagy through COL5A1 and FSTL1, impacting DDP resistance.
Conclusions:
- MiR-215-5p targets COL5A1 to regulate FSTL1 and autophagy, thereby overcoming DDP resistance in LUAD.
- This study enhances understanding of LUAD pathogenesis and offers potential therapeutic targets for overcoming drug resistance.
