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Preparation and In Vitro Characterization of Magnetized miR-modified Endothelial Cells
Published on: May 2, 2017
miR‑146a‑5p targets TCSF and influences cell growth and apoptosis to repress NSCLC progression
Wen-Ting Huang1, Rong-Quan He2, Xiao-Jiao Li3
1Department of Pathology, First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region 530021, P.R. China.
Abstract:
Several studies have indicated that microRNAs (miRs) mediate multiple pathways associated with tumorigenesis and progression. Our preliminary study experimentally verified that miR‑146a‑5p has a role in the biological behavior of non‑small cell lung cancer (NSCLC) cells. To perform further investigation of miR‑146a‑5p, the present study evaluated miR‑146a‑5p by targeting its downstream gene tumor collagenase stimulatory factor (TCSF) to influence cell viability, proliferation and apoptosis in NSCLC. Online sequence prediction, a thorough search of the open source database The Cancer Genome Atlas (TCGA), immunohistochemistry (IHC) of TCSF in clinical lung cancer tissues, and a dual‑luciferase assay, as well as assays to test viability, proliferation and apoptosis in vitro, were conducted to explain the targeted regulation association between miR‑146a‑5p and TCSF in NSCLC. The miRanda and TargetScanHuman database revealed that TCSF and miR‑146a‑5p had target binding sites. A luciferase reporter assay demonstrated that miR‑146a‑5p and TCSF did have complementary sequences (P<0.05). From the TCGA database, TCSF was highly expressed in lung adenocarcinoma and lung squamous cell carcinoma tissues when compared with normal lung tissues (P<0.05). Furthermore, the protein level of TCSF in cancerous lung tissues was determined by IHC, and it was concluded that TCSF protein was also upregulated in NSCLC tissues (P<0.001). A significant difference was identified following in vitro experiments for the NSCLC cell line A549, which revealed that miR‑146a‑5p and TCSF regulated cell viability, proliferation and apoptosis. In conclusion, the present study verified the target action association between TCSF and miR‑146a‑5p with high throughput data analysis and experimental results in NSCLC.
Insights
MicroRNAs (miRs) regulate non-small cell lung cancer (NSCLC) progression. This study confirms miR-146a-5p targets tumor collagenase stimulatory factor (TCSF), impacting NSCLC cell viability, proliferation, and apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- MicroRNAs (miRs) are key regulators in tumorigenesis and cancer progression.
- miR-146a-5p has been implicated in the biological behavior of non-small cell lung cancer (NSCLC) cells.
Purpose of the Study:
- To investigate the role of miR-146a-5p in NSCLC by examining its targeting of the downstream gene tumor collagenase stimulatory factor (TCSF).
- To determine the influence of the miR-146a-5p/TCSF interaction on NSCLC cell viability, proliferation, and apoptosis.
Main Methods:
- Utilized online databases (miRanda, TargetScanHuman, TCGA) for sequence prediction and expression analysis.
- Performed dual-luciferase reporter assays to confirm direct targeting.
- Conducted immunohistochemistry (IHC) on clinical lung cancer tissues.
- Carried out in vitro assays to assess cell viability, proliferation, and apoptosis in NSCLC cell lines.
Main Results:
- TCSF was identified as a direct target of miR-146a-5p.
- TCSF exhibited high expression in lung adenocarcinoma and lung squamous cell carcinoma tissues compared to normal tissues.
- TCSF protein levels were upregulated in NSCLC tissues.
- miR-146a-5p and TCSF significantly regulated cell viability, proliferation, and apoptosis in vitro.
Conclusions:
- The study verified the targeted regulatory association between miR-146a-5p and TCSF in NSCLC.
- This interaction plays a significant role in the biological behavior of NSCLC cells, affecting viability, proliferation, and apoptosis.
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