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Induction of Mesenchymal-Epithelial Transitions in Sarcoma Cells
Published on: April 7, 2017
MicroRNA-200a inhibits epithelial-mesenchymal transition in human hepatocellular carcinoma cell line
Chong Zhong1, Ming-Yi Li2, Zhi-Yuan Chen3
1Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine Guangzhou 510405, P. R. China.
Objective:
Our study investigated the role of microRNA (miR)-200a and its molecular targets in hepatocellular carcinoma (HCC) cells.
Methods:
An inhibitor of miR-200a was transiently transfected into the hepatocellular carcinoma cell line, MHCC-97L. The effect of this transfection on mRNA levels of epithelial-mesenchymal transition (EMT)-related genes was measured by fluorescence-based quantitative real-time polymerase chain reaction (qRT-PCR). Further, protein levels of EMT-related genes, cell proliferation and apoptosis-related markers were assessed by Western blot analysis in these transfected cells. MTT and wound-healing assay were used to evaluate the proliferation and migration of MHCC-97L cells in presence and in absence of miR-200a inhibitor.
Results:
Compared with miR-NC control group, qRT-PCR results in anti-miR-200a group revealed a significant reduction in the mRNA levels of E-cadherin, with a concomitant increasing in vimentin mRNA level (all P < 0.05). Western blot results showed higher E-cadherin and Caspase-3 protein expressions in anti-miR-200a group compared to miR-NC group (P < 0.05). In addition, vimentin and Ki-67 protein expression was found sharply decreased in anti-miR-200a group compared to miR-NC group (P < 0.05). Consistent with this, wound-healing and MTT assay showed that migration and proliferation capacity of MHCC-97L cells in anti-miR-200a group is significantly increased compared with miR-NC group (both P < 0.05).
Conclusion:
Our study reveals an important role of miR-200a in inhibiting EMT, proliferation and migration in HCC cells, suggesting the possibility of miR-200a-based therapeutics in HCC.
Insights
MicroRNA-200a inhibits hepatocellular carcinoma (HCC) progression by suppressing epithelial-mesenchymal transition (EMT), proliferation, and migration. This suggests miR-200a as a potential therapeutic target for HCC treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- MicroRNAs (miRNAs) play critical roles in cancer development and progression.
- The specific role of miR-200a in HCC requires further elucidation.
Purpose of the Study:
- To investigate the function of miR-200a in hepatocellular carcinoma (HCC) cells.
- To identify the molecular targets of miR-200a involved in HCC progression.
- To explore the potential of miR-200a as a therapeutic agent for HCC.
Main Methods:
- Transfection of miR-200a inhibitor into MHCC-97L HCC cells.
- Quantitative real-time PCR (qRT-PCR) to assess mRNA levels of epithelial-mesenchymal transition (EMT)-related genes.
- Western blot analysis for protein expression of EMT markers, proliferation, and apoptosis.
- MTT and wound-healing assays to evaluate cell proliferation and migration.
Main Results:
- Inhibition of miR-200a led to decreased E-cadherin and increased vimentin mRNA levels.
- Protein analysis revealed increased E-cadherin and Caspase-3, with decreased vimentin and Ki-67.
- MHCC-97L cells with miR-200a inhibition showed significantly increased proliferation and migration.
Conclusions:
- miR-200a plays a crucial role in suppressing EMT, proliferation, and migration in HCC cells.
- Downregulation of miR-200a promotes HCC cell aggressiveness.
- miR-200a holds promise as a potential therapeutic strategy for HCC.
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The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...