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Updated: Nov 29, 2025

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
miR-141 Promotes Colon Cancer Cell Proliferation by Targeted PHLPP2 Expression Inhibitionn
Fazhuang Fang1, Ling Cheng2, Xiaotang Wu2
1Department of Hepatobiliary, Pancreatic and Gastric Surgery, Jinhua Guangfu Hospital, Jinhua 321000, People's Republic of China.
Objective:
Colon cancer (CC) is the third most common cancer with a high rate of incidence and mortality. Therefore, it is highly necessary to explore novel targets of CC.
Methods:
The miRNA-seq and RNA-seq data of CC were accessed from the TCGA database. Differential analysis was performed using the "edgeR" package to identify differentially expressed miRNAs (DE_miRNAs). The downstream target genes of the target miRNA were then predicted by miRNA target prediction databases to identify the target mRNA. Normal colon cell line CCD-18Co and CC cell lines HCT-116, HT-29, SW620 and SW480 were chosen, and qRT-PCR was conducted to detect miR-141 expression in these cell lines. qRT-PCR and Western blot were carried out to determine PHLPP2 mRNA and protein expression, respectively. Dual-luciferase reporter gene assay was performed to verify the targeting relationship between miR-141 and PHLPP2 3'UTR. CCK-8 assay and colony formation assay were carried out to detect cell proliferation. Meanwhile, tumor xenograft model in nude mice was constructed to assess CC cell tumorigenic ability in vivo.
Results:
miR-141 was markedly up-regulated in CC tissue. CC cell proliferation and in vivo tumorigenic ability were suppressed by miR-141 silencing but promoted by miR-141 over-expression. PHLPP2 was significantly down-regulated in cancer tissue. Dual-luciferase reporter gene assay indicated that miR-141 could bind to PHLPP2 3'UTR. PHLPP2 expression was noticeably elevated upon miR-141 deficiency but significantly inhibited upon miR-141 over-expression. CCK-8 and colony formation assay suggested that miR-141 facilitated CC cell proliferation by silencing PHLPP2.
Conclusion:
miR-141 promotes CC cell proliferation by targeted silencing PHLPP2.
Insights
MicroRNA-141 (miR-141) promotes colon cancer (CC) cell proliferation by targeting and silencing PH-like गर्नुहोस् protein phosphatase 2 (PHLPP2). This finding identifies a novel therapeutic target for colon cancer treatment.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- Colon cancer (CC) is a leading cause of cancer incidence and mortality.
- Identifying novel therapeutic targets is crucial for effective CC treatment.
- MicroRNAs play significant roles in cancer development and progression.
Purpose of the Study:
- To investigate the role of miR-141 in colon cancer.
- To identify the downstream target of miR-141 involved in CC.
- To explore the therapeutic potential of targeting the miR-141/PHLPP2 axis in CC.
Main Methods:
- Analysis of miRNA-seq and RNA-seq data from The Cancer Genome Atlas (TCGA) database.
- Differential expression analysis using edgeR to identify differentially expressed miRNAs (DE_miRNAs).
- Validation of miR-141 and PHLPP2 expression in CC cell lines using qRT-PCR and Western blot.
- Dual-luciferase reporter gene assay to confirm the targeting relationship between miR-141 and PHLPP2.
- In vitro (CCK-8, colony formation assays) and in vivo (tumor xenograft model) experiments to assess cell proliferation and tumorigenic ability.
Main Results:
- miR-141 was significantly upregulated in colon cancer tissues and cell lines.
- miR-141 silencing suppressed CC cell proliferation and in vivo tumor growth, while overexpression promoted them.
- PHLPP2 was downregulated in CC tissues, and miR-141 directly targeted and silenced PHLPP2.
- Restoration of PHLPP2 expression inhibited miR-141-induced CC cell proliferation.
Conclusions:
- miR-141 acts as an oncomiR in colon cancer by promoting cell proliferation.
- The oncogenic role of miR-141 is mediated through the targeted silencing of PHLPP2.
- The miR-141/PHLPP2 pathway represents a potential therapeutic target for colon cancer.
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