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Published on: September 12, 2019
miR-204-5p suppresses hepatocellular cancer proliferation by regulating homeoprotein SIX1 expression
Yi Chu1, Mingzuo Jiang1, Feng Du1
1State Key Laboratory of Cancer Biology & Institute of Digestive Diseases Xijing Hospital The Fourth Military Medical University Xi'an China.
Abstract:
Fewer than 30% of patients with hepatocellular carcinoma (HCC) are eligible to receive curative therapies, and so a better understanding of the molecular mechanisms of HCC is needed to identify potential therapeutic targets. The role of microRNA (miRNA) in modulating tumour progression has been demonstrated, and therapies targeting miRNA appear promising. miR-204-5p has been shown to function in numerous types of cancer, but its role in HCC remains unclear. In this study, we found that miR-204-5p expression was downregulated in cancerous HCC tissues compared to nontumour tissues. Kaplan-Meier survival curve analysis also showed that low expression of miR-204-5p predicted worse outcomes of HCC patients. In addition, miR-204-5p expression was significantly lower in HCC cell lines. The function of miR-204-5p was also assessed both in vitro and in vivo. We demonstrated that ectopic expression of miR-204-5p in HCC cell lines inhibited HCC cell proliferation and clonogenicity using CCK8, BrdU and colony-forming assays, while the inhibition of miR-204-5p enhanced proliferation and clonogenicity. Further in vivo studies in mice further confirmed the proliferation capacity of miR-204-5p. We also identified sine oculis homeobox homologue 1 (SIX1) as a direct target of miR-204-5p and showed that it was inversely correlated with miR-204-5p in both human and mouse HCC tissues. Transfection of miR-204-5p mimics in BEL-7404 cells blocked the cell cycle by inhibiting the expression of cyclin-D1 and cyclin-A1, cell cycle-related factors regulated by SIX1. More importantly, overexpression of the 3'UTR mutant SIX1 but not the wild-type SIX1 abolished the suppressive effect of miR-204-5p, and downregulated SIX1 in BEL-7402 cells that transfected with miR-204 inhibitors could partly block the inhibitory effect of miR-204-5p on proliferation. Thus, we have demonstrated that miR-204-5p suppresses HCC proliferation by directly regulating SIX1 and its downstream factors.
Insights
MicroRNA-204-5p (miR-204-5p) is downregulated in hepatocellular carcinoma (HCC), suppressing tumor growth by targeting SIX1. Low miR-204-5p predicts poor HCC patient outcomes, highlighting its therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Hepatocellular carcinoma (HCC) has limited curative treatment options, necessitating research into molecular mechanisms for novel therapeutic targets.
- MicroRNAs (miRNAs) are recognized regulators of tumor progression, with miRNA-targeted therapies showing promise.
- The specific role of miR-204-5p in HCC pathogenesis remains largely uncharacterized.
Purpose of the Study:
- To investigate the expression levels and functional role of miR-204-5p in hepatocellular carcinoma.
- To identify the molecular targets and pathways regulated by miR-204-5p in HCC.
- To evaluate the potential of miR-204-5p as a therapeutic agent for HCC.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to assess miR-204-5p expression in HCC tissues and cell lines.
- In vitro assays (CCK8, BrdU, colony-forming assays) to evaluate the effect of miR-204-5p on HCC cell proliferation and clonogenicity.
- In vivo mouse models to confirm the role of miR-204-5p in tumor growth.
- Luciferase reporter assays and Western blotting to validate SIX1 as a direct target of miR-204-5p.
- Cell cycle analysis to determine the impact of miR-204-5p/SIX1 axis on cell cycle progression.
Main Results:
- miR-204-5p expression was significantly downregulated in HCC tissues and cell lines compared to non-tumorous tissues.
- Low miR-204-5p expression correlated with poorer patient survival.
- Ectopic expression of miR-204-5p inhibited HCC cell proliferation and clonogenicity, while inhibition of miR-204-5p promoted these processes.
- SIX1 was identified as a direct target of miR-204-5p, with inverse correlation in HCC tissues.
- miR-204-5p suppressed HCC cell cycle progression by inhibiting SIX1 and its downstream effectors, cyclin-D1 and cyclin-A1.
Conclusions:
- miR-204-5p functions as a tumor suppressor in hepatocellular carcinoma.
- The tumor-suppressive activity of miR-204-5p is mediated through direct inhibition of SIX1 and subsequent downregulation of cell cycle regulators.
- miR-204-5p represents a potential biomarker for HCC prognosis and a promising therapeutic target.
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