Lnc-TMEM132D-AS1 overexpression reduces sensitivity of non-small cell lung cancer cells to osimertinib.
10.12122/j.issn.1673-4254.2023.02.12
- Author:
Qi Lin ZHAO
1
;
Nan WANG
2
;
Ya Wen LI
2
;
Qing Tan WU
1
;
Lan Xiang WU
2
Author Information
1. Department of Cardiothoracic Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
2. Institute of Life Sciences, Chongqing Medical University, Chongqing 400016, China.
- Publication Type:Journal Article
- Keywords:
acquired drug resistance;
lnc-TMEM132D-AS1;
long non-coding RNA;
non-small cell lung cancer;
osimertinib
- MeSH:
Humans;
Carcinoma, Non-Small-Cell Lung/metabolism*;
Lung Neoplasms/genetics*;
RNA, Long Noncoding/metabolism*;
Sincalide/metabolism*;
Cell Line, Tumor;
Cell Proliferation/genetics*;
Cell Movement;
MicroRNAs/genetics*;
Gene Expression Regulation, Neoplastic;
Membrane Proteins/metabolism*
- From:
Journal of Southern Medical University
2023;43(2):242-250
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To screen the differentially expressed long non-coding RNAs (lncRNAs) in non-small cell lung cancer (NSCLC) cells with acquired resistance to osimertinib and explore their roles in drug resistance of the cells.
METHODS:The cell lines H1975_OR and HCC827_OR with acquired osimertinib resistance were derived from their osimertinib-sensitive parental NSCLC cell lines H1975 and HCC827, respectively, and their sensitivity to osimertinib was assessed with CCK-8 assay, clone formation assay and flow cytometry. RNA sequencing (RNA-seq) and real-time quantitative PCR (qPCR) were used to screen the differentially expressed lncRNAs in osimertinib-resistant cells. The role of the identified lncRNA in osimertinib resistance was explored using CCK-8, clone formation and Transwell assays, and its subcellular localization and downstream targets were analyzed by nucleoplasmic separation, bioinformatics analysis and qPCR.
RESULTS:The resistance index of H1975_OR and HCC827_OR cells to osimertinib was 598.70 and 428.82, respectively (P < 0.001), and the two cell lines showed significantly increased proliferation and colony-forming abilities with decreased apoptosis (P < 0.01). RNA-seq identified 34 differentially expressed lncRNAs in osimertinib-resistant cells, and among them lnc-TMEM132D-AS1 showed the highest increase of expression after acquired osimertinib resistance (P < 0.01). Analysis of the TCGA database suggested that the level of lnc-TMEM132D-AS1 was significantly higher in NSCLC than in adjacent tissues (P < 0.001), and its high expression was associated with a poor prognosis of the patients. In osimertinib-sensitive cells, overexpression of Lnc-TMEM132D-AS1 obviously promoted cell proliferation, colony formation and migration (P < 0.05), while Lnc-TMEM132D-AS1 knockdown partially restored osimertinib sensitivity of the resistant cells (P < 0.01). Lnc-TMEM132D-AS1 was localized mainly in the cytoplasm, and bioinformatics analysis suggested that hsa-miR-766-5p was its candidate target, and their expression levels were inversely correlated. The target mRNAs of hsa-miR-766-5p were mainly enriched in the Ras signaling pathway.
CONCLUSION:The expression of lnc-TMEM132D-AS1 is significantly upregulated in NSCLC cells with acquired osimertinib resistance, and may serve as a potential biomarker and therapeutic target for osimertinibresistant NSCLC.
