Related Experiment Video
Updated: Dec 9, 2025
Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer
Published on: March 14, 2019
MicroRNA-421 Inhibits Apoptosis by Downregulating Caspase-3 in Human Colorectal Cancer
Yifan Zhou1, Xiaowen Cheng1,2, Yufeng Wan3
1Department of Clinical Laboratory, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, People's Republic of China.
Purpose:
Dysregulated microRNAs (miRNAs/miRs) have been reported to play significant roles in pathogenesis of colorectal cancer (CRC). Previous studies have demonstrated that miR-421 regulates apoptosis in some cancers. Caspase-3 plays a key role in apoptosis, but the relationship between miR-421 and caspase-3 in CRC has not been determined. In this study, we investigated the role of miR-421 in CRC and the relationship between miR-421 and caspase-3.
Methods:
Expression of miR-421 and caspase-3 were detected in human paired CRC cancer tissues and corresponding paracancerous tissues. In situ detection of tissue, apoptosis was performed via the TUNEL assay. HCT116 and SW480 cell lines were subjected to several in vitro experiments to explore the relationship between miRNA421 and caspase-3 during apoptosis using miR421 mimics/antagomir and luciferase reporter assay. Apoptosis was measured by determining the levels and activity of caspase-3 as well as DNA fragmentation. Luciferase reporter assay was performed to determine the potential interaction of miR-421 with caspase-3.
Results:
The results showed that the expression of miR-421 in cancer tissues was higher than that in corresponding paracancerous tissues. Inhibition of miR-421 induced apoptosis, as shown by the upregulation of caspase-3 activity and expression as well as DNA fragmentation, which were attenuated by miR-421 mimic. We further showed that miR-421 targeted and inhibited CASP3 expression by targeting sites located in the 3'-untranslated region (3'-UTR) of CASP3 mRNA.
Conclusion:
This study demonstrated an anti-apoptotic role of miR-421 in CRC and identified caspase-3 gene as a direct target of miR-421. These findings provide a potential treatment strategy using miR-421 as a therapeutic target for CRC.
Insights
MicroRNA-421 (miR-421) promotes colorectal cancer (CRC) by inhibiting apoptosis. This study found miR-421 targets and downregulates caspase-3, suggesting miR-421 as a potential therapeutic target for CRC.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- MicroRNAs (miRNAs) are implicated in colorectal cancer (CRC) pathogenesis.
- miR-421 is known to regulate apoptosis in various cancers.
- The specific role of miR-421 and its interaction with caspase-3 in CRC remains unclear.
Purpose of the Study:
- To investigate the role of miR-421 in colorectal cancer.
- To determine the relationship between miR-421 and caspase-3 in CRC apoptosis.
- To explore miR-421 as a potential therapeutic target for CRC.
Main Methods:
- Detected miR-421 and caspase-3 expression in CRC tissues and cell lines.
- Utilized TUNEL assay for in situ apoptosis detection.
- Performed in vitro experiments with miR-421 mimics/antagomirs and luciferase reporter assays.
- Assessed apoptosis by measuring caspase-3 activity and DNA fragmentation.
Main Results:
- miR-421 expression was significantly higher in CRC tissues compared to adjacent tissues.
- Inhibition of miR-421 led to increased apoptosis, evidenced by upregulated caspase-3 and DNA fragmentation.
- miR-421 directly targets and inhibits the CASP3 gene by binding to its 3'-untranslated region.
Conclusions:
- miR-421 exhibits an anti-apoptotic function in colorectal cancer.
- Caspase-3 (CASP3) is identified as a direct downstream target of miR-421.
- Targeting miR-421 presents a potential therapeutic strategy for colorectal cancer treatment.
Related Concept Videos
01:22MicroRNAs
01:22MicroRNAs
01:31The Intrinsic Apoptotic Pathway
02:15Experimental RNAi