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An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA-155-5p suppresses PD-L1 expression in lung adenocarcinoma
Jiansheng Huang1, Qiaoyou Weng1, Yang Shi1
1Key Laboratory of Imaging Diagnosis and Minimally Invasive Intervention Research, Lishui Hospital of Zhejiang University, China.
Abstract:
MiR-155-5p is a key oncogenic microRNA that maintains immune homeostasis and mediates cross-talk between inflammation and tumorigenesis. High expression of programmed death ligand-1 (PD-L1) also plays an important role in immune tolerance in tumors. The present study aimed to explore the relationship between miR-155-5p and PD-L1 in lung adenocarcinoma (LUAD) cells A549 and H1650. The expression levels of miR-155-5p and PD-L1 in LUAD patients were detected by a quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and mimics of miR-155-5p were used to model increased expression in A549 or H1650 cells. After 24 h, we measured levels of PD-L1 by qRT-PCR, western blotting and flow cytometry. In addition, we identified two sites in the PD-L1 3'-UTR (5'-AGCAUUA-3' and 5'-GCAUUAA-3') that can be bound by miR-155-5p using TargetScan (http://www.targetscan.org). Compared to normal tissue, miR-155-5p was overexpressed in tumor tissue (P = 0.0456), whereas the expression of PD-L1 was not significantly different (P = 0.1349). The expression levels of miR-155-5p and PD-L1 were negatively correlated (r = -0.6409, P = 0.0459 and r = -0.7544, P = 0.0117). Exogenous overexpression of miR-155-5p decreased the mRNA, total protein and membrane protein expression levels of PD-L1 both in A549 and H1650 cells (P < 0.05). Taken together, our data suggest that miR-155-5p may suppress the expression of PD-L1 in LUAD.
Insights
MicroRNA-155-5p (miR-155-5p) may suppress programmed death ligand-1 (PD-L1) in lung adenocarcinoma. This study found miR-155-5p overexpression negatively correlates with PD-L1 levels in LUAD cells, suggesting a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- MicroRNA-155-5p (miR-155-5p) is an oncogenic microRNA involved in immune homeostasis and cancer development.
- Programmed death ligand-1 (PD-L1) is crucial for immune tolerance in tumors, often overexpressed in cancer.
- Understanding the interplay between miR-155-5p and PD-L1 is vital for lung adenocarcinoma (LUAD) research.
Purpose of the Study:
- To investigate the relationship between miR-155-5p and PD-L1 expression in LUAD.
- To determine if miR-155-5p influences PD-L1 levels in LUAD cell lines.
Main Methods:
- Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) to measure miR-155-5p and PD-L1 expression.
- Transfection with miR-155-5p mimics in A549 and H1650 LUAD cells.
- Western blotting and flow cytometry to assess PD-L1 protein levels.
Main Results:
- miR-155-5p was overexpressed in LUAD tumor tissue compared to normal tissue.
- A significant negative correlation was observed between miR-155-5p and PD-L1 expression levels.
- Overexpression of miR-155-5p led to decreased mRNA, total protein, and membrane protein expression of PD-L1 in LUAD cells.
Conclusions:
- miR-155-5p expression is inversely correlated with PD-L1 expression in lung adenocarcinoma.
- miR-155-5p may act as a suppressor of PD-L1 expression in LUAD.
- These findings suggest a potential role for miR-155-5p in modulating the tumor immune microenvironment in LUAD.
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