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Updated: Apr 1, 2026

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA‑144 inhibits migration and proliferation in rectal cancer by downregulating ROCK‑1
Shang-Dang Cai1, Jian-She Chen2, Zuo-Wu Xi1
1Anorectal Branch, Henan Province Hospital of TCM, Zhengzhou, Henan 450002, P.R. China.
Abstract:
Cancer of the colon and rectum are two distinct entities, which require different treatment strategies and separate treatment. MicroRNAs (miRNAs) act as critical regulators of genes involved in several biological processes. Aberrant alterations of miRNAs have been found in several types of cancer, including colon cancer and rectal cancer. Extensive catalogues of downregulated miRNAs have been identified for colon cancer, whereas only limited data are available for rectal cancer. An example of miRNA profiling in a previous study found that miRNA (miR)‑144 showed aberrant expression and appeared to be rectal cancer‑specific, its expression not being reported in colon cancer. In the present study, the role of miR‑144 in rectal cancer was investigated. SW837 and SW1463 cell lines were selected as rectal cell carcinoma cells. Using reverse transcription-quantitative polymerase chain reaction, western blot, BrdU, cell migration and cell viability assays, it was found that the expression levels of miR‑144 were significantly reduced in the SW837 and SW1463 cell lines, and the overexpression of miR‑144 suppressed rectal cancer cell viability, migration and proliferation. In addition, Rho‑associated coiled‑coil containing protein kinase 1 (ROCK1) was identified as a target of miR‑144 in the rectal cancer cells. The supplementation of ROCK1 markedly restored the cell migration and proliferation, which was inhibited by miR‑144. Together, the data of the present study demonstrated that miR‑144 acts as a tumor suppressor by targeting ROCK1, and indicates the potential of miR‑144 as a novel biomarker and target in the treatment of rectal cancer.
Insights
MicroRNA-144 (miR-144) functions as a tumor suppressor in rectal cancer. This study found miR-144 inhibits rectal cancer cell viability, migration, and proliferation by targeting ROCK1, suggesting its potential as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Rectal and colon cancers are distinct, requiring different treatments.
- MicroRNAs (miRNAs) regulate genes and are implicated in cancer development.
- Limited data exists on miRNA alterations in rectal cancer compared to colon cancer.
Purpose of the Study:
- To investigate the role of microRNA-144 (miR-144) in rectal cancer.
- To identify potential molecular targets of miR-144 in rectal cancer cells.
Main Methods:
- Quantitative polymerase chain reaction and western blot to assess miR-144 and ROCK1 expression.
- Cell viability, proliferation (BrdU), and migration assays were performed.
- Functional rescue experiments with Rho-associated coiled-coil containing protein kinase 1 (ROCK1) supplementation.
Main Results:
- miR-144 expression was significantly reduced in rectal cancer cell lines (SW837, SW1463).
- Overexpression of miR-144 suppressed rectal cancer cell viability, migration, and proliferation.
- ROCK1 was identified as a direct target of miR-144, and its restoration reversed miR-144's inhibitory effects.
Conclusions:
- miR-144 acts as a tumor suppressor in rectal cancer by targeting ROCK1.
- miR-144 demonstrates potential as a novel biomarker and therapeutic target for rectal cancer treatment.
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