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NAT10 Knockdown Improves Cisplatin Sensitivity in Non-Small Cell Lung Cancer by Inhibiting the TRIM44/PI3K/AKT
Qi Sun1, Xiansong Yang2, Ye Wang3
1Department of Thoracic Surgery, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.
Background:
Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related deaths worldwide, and cisplatin (DDP) resistance remains a significant challenge in NSCLC treatment.
Methods:
Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to analyze NAT10 and tripartite motif containing 44 (TRIM44) mRNA levels. Western blotting assay was used to detect protein expression. Cell viability was analyzed by a cell counting kit-8 assay. Cell proliferation, apoptosis, invasion, and stem-like traits were assessed using a 5-Ethynyl-2'-deoxyuridineassay, flow cytometry, Transwell invasion assay, and sphere formation assay, respectively. The association between NAT10 and TRIM44 was identified by an RNA immunoprecipitation assay. A xenograft mouse model was established to evaluate the effect of NAT10 silencing on DDP sensitivity in vivo.
Results:
NAT10 expression was upregulated in DDP-resistant NSCLC tissues and cells. NAT10 knockdown enhanced DDP sensitivity in DDP-resistant NSCLC cells, accompanied by decreased protein expression of multidrug resistance 1 (MDR1). The silencing of NAT10 also inhibited the proliferation, invasion, and stem-like traits of DDP-resistant NSCLC cells, while inducing cell apoptosis. However, NAT10 overexpression displayed the opposite effects. Moreover, NAT10 maintained TRIM44 mRNA stability in an ac4C-dependent manner. TRIM44 overexpression reversed the NAT10 knockdown-induced effects on DDP sensitivity and the malignant progression of NSCLC cells. In addition, NAT10 silencing inactivated the PI3K/AKT pathway by regulating TRIM44 in DDP-resistant NSCLC cells. The treatment of the PI3K/AKT pathway inhibitor, LY294002, mitigated the effects of TRIM44 overexpression on DDP sensitivity and NSCLC cell progression. Further, NAT10 knockdown improved the sensitivity of tumors to DDP in vivo.
Conclusion:
NAT10 knockdown improved DDP sensitivity in NSCLC by inhibiting the TRIM44/PI3K/AKT pathway, which may have significant clinical implications for overcoming DDP resistance in NSCLC treatment.
Insights
Reducing NAT10 levels enhances cisplatin sensitivity in non-small cell lung cancer (NSCLC) by inhibiting the TRIM44/PI3K/AKT pathway. This finding offers a potential strategy to overcome drug resistance in NSCLC treatment.
Area of Science:
- Molecular Oncology
- Cancer Therapeutics
- Drug Resistance Mechanisms
Background:
- Non-small cell lung cancer (NSCLC) is a major global health concern, with cisplatin (DDP) resistance posing a significant therapeutic challenge.
- Understanding the molecular mechanisms underlying DDP resistance is crucial for developing effective treatment strategies.
Purpose of the Study:
- To investigate the role of NAT10 in cisplatin resistance in NSCLC.
- To elucidate the molecular pathway through which NAT10 influences DDP sensitivity and cancer progression.
Main Methods:
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting were used to assess NAT10 and TRIM44 expression.
- Cell viability, proliferation, apoptosis, invasion, and stem-like traits were evaluated using CCK-8, EdU, flow cytometry, Transwell, and sphere formation assays.
- RNA immunoprecipitation (RIP) assays and in vivo xenograft models were employed to determine the NAT10-TRIM44 interaction and therapeutic efficacy.
Main Results:
- NAT10 was upregulated in DDP-resistant NSCLC, and its knockdown resensitized cells to DDP, reducing multidrug resistance 1 (MDR1) expression.
- NAT10 silencing inhibited NSCLC cell proliferation, invasion, and stem-like properties while promoting apoptosis; NAT10 overexpression had opposite effects.
- NAT10 stabilized TRIM44 mRNA, and TRIM44 overexpression counteracted the effects of NAT10 knockdown; NAT10 silencing inactivated the PI3K/AKT pathway via TRIM44.
Conclusions:
- NAT10 knockdown enhances DDP sensitivity in NSCLC by inhibiting the TRIM44/PI3K/AKT pathway.
- Targeting NAT10 presents a promising therapeutic strategy to overcome cisplatin resistance in NSCLC.
- These findings have significant clinical implications for improving NSCLC treatment outcomes.
