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Updated: Jun 22, 2026
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MicroRNA-9 reduces cell invasion and E-cadherin secretion in SK-Hep-1 cell
Hao-Xiang Tan1, Qian Wang, Lian-Zhou Chen
1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, 510080, China.
Abstract:
MicroRNAs (miRNAs) are an abundant class of short noncoding RNAs that can posttranscriptionally regulate gene expression in animals. They are also involved in cancer initiation and progression, and their expression profiles serve as phenotypic signatures of different cancers. The roles played by microRNAs specifically in "micromanagement of metastasis" has been addressed only recently. The molecular mechanisms of hepatocellular carcinoma (HCC) metastasis are still poorly understood. Recent evidence implies genetic determinants of cancer metastasis. Because gene expression signature significantly differs between primary metastasis-free HCC and primary HCC with intrahepatic metastases, miRNA expression in those primary HCC may change correspondingly. The 28 up-regulated miRNAs, part of the reported miRNA profiles of HCC, were compared in primary HCC with or without metastases. Only eight miRNAs were found to be significantly up-regulated in primary HCC with metastases while miR-9 had the highest hold change. miR-9 was highly expressed in SK-Hep-1 cell when compared with other hepatoma cell lines and downregulation of miR-9 reduced SK-Hep-1 cell invasion. E-cadherin, a tumor invasion suppressor in HCC, was found to be a putative gene target of miR-9. E-cadherin was up-regulated by miR-9 inhibitor. The findings suggest miR-9 could be involved in HCC metastasis.
Insights
MicroRNAs (miRNAs) regulate gene expression and are implicated in cancer. This study found miR-9 is highly expressed in hepatocellular carcinoma (HCC) with metastasis, suggesting its role in cancer spread.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- MicroRNAs (miRNAs) are short noncoding RNAs regulating gene expression posttranscriptionally.
- miRNA expression profiles are cancer-specific and play roles in cancer initiation, progression, and metastasis.
- The molecular mechanisms of hepatocellular carcinoma (HCC) metastasis are not fully understood, but genetic factors are implicated.
Purpose of the Study:
- To investigate the role of specific microRNAs in the metastasis of hepatocellular carcinoma (HCC).
- To identify differentially expressed miRNAs in primary HCC with and without intrahepatic metastases.
- To explore the functional role of miR-9 and its potential targets in HCC metastasis.
Main Methods:
- Comparison of miRNA expression profiles between primary HCC with and without metastases.
- Quantitative analysis of specific miRNA expression in HCC cell lines.
- Functional assays to assess the effect of miR-9 modulation on cell invasion.
- Bioinformatic prediction and experimental validation of miR-9 gene targets.
Main Results:
- Eight miRNAs were significantly up-regulated in primary HCC with metastases compared to metastasis-free HCC.
- miR-9 showed the highest fold change among the up-regulated miRNAs.
- High miR-9 expression in SK-Hep-1 cells correlated with increased invasion; its downregulation reduced invasion.
- E-cadherin, a known tumor suppressor, was identified as a putative target of miR-9 and was upregulated by a miR-9 inhibitor.
Conclusions:
- miR-9 is significantly up-regulated in metastatic HCC and promotes cell invasion.
- miR-9 may contribute to HCC metastasis by down-regulating E-cadherin expression.
- Targeting miR-9 could be a potential therapeutic strategy for inhibiting HCC metastasis.
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