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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
miR-137 inhibits the proliferation of human non-small cell lung cancer cells by targeting SRC3
Ruilin Chen1, Yongqing Zhang1, Chengcheng Zhang1
1Department of Respiratory Medicine, Shaanxi Provincial People's Hospital, Xi'an, Shaanxi 710068, P.R. China.
Abstract:
Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. The results of the present study demonstrate that high expression of microRNA (miR)-137 and low expression of steroid receptor coactivator-3 (SRC3) had a significant negative correlation in 40 NSCLC tissue samples. In addition, cell colony formation and proliferation was significantly reduced in miR-137-transfected A549 and NCI-H838 cells compared with scramble-transfected NSCLC cell lines. miR-137 was identified to induce G1/S cell cycle arrest and dysregulate the mRNA expression of cell cycle-associated proteins (proliferating cell nuclear antigen, cyclin E, cyclin A1, cyclin A2 and p21) in NSCLC cells. Notably, miR-137 could significantly suppress SRC3 3' untranslated region (UTR) luciferase-reporter activity, an effect that was not detectable when the putative 3'-UTR target-site was mutated, further clarifying the molecular mechanisms underlying the role of miR-137 in NSCLC. In conclusion, the results of the present study suggest that miR-137 suppresses NSCLC cell proliferation by partially targeting SRC3.
Insights
High microRNA (miR)-137 expression correlates with low steroid receptor coactivator-3 (SRC3) in non-small cell lung cancer (NSCLC). miR-137 suppresses NSCLC cell proliferation by targeting SRC3, offering potential therapeutic insights.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Non-small cell lung cancer (NSCLC) is the most prevalent form of lung cancer.
- MicroRNAs (miRNAs) play crucial roles in cancer development and progression.
- Steroid receptor coactivator-3 (SRC3) is implicated in various cellular processes, including proliferation.
Purpose of the Study:
- To investigate the correlation between microRNA (miR)-137 and steroid receptor coactivator-3 (SRC3) expression in NSCLC.
- To elucidate the functional role of miR-137 in NSCLC cell proliferation and cell cycle regulation.
- To identify the molecular mechanism by which miR-137 affects NSCLC.
Main Methods:
- Correlation analysis of miR-137 and SRC3 expression in 40 NSCLC tissue samples.
- In vitro experiments involving miR-137 transfection in NSCLC cell lines (A549, NCI-H838).
- Assessment of cell proliferation, colony formation, cell cycle progression, and luciferase-reporter assays to validate target interaction.
Main Results:
- A significant negative correlation was observed between high miR-137 and low SRC3 expression in NSCLC tissues.
- miR-137 transfection significantly reduced NSCLC cell proliferation and colony formation.
- miR-137 induced G1/S cell cycle arrest and altered the expression of key cell cycle regulators.
- miR-137 directly suppressed SRC3 3' untranslated region (UTR) activity, confirming SRC3 as a target.
Conclusions:
- miR-137 acts as a tumor suppressor in NSCLC by inhibiting cell proliferation.
- The suppressive effect of miR-137 on NSCLC is partially mediated through the targeting of SRC3.
- These findings highlight the potential of miR-137 as a therapeutic target for NSCLC.
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