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MST4 Regulates Epithelial-Mesenchymal Transition of Choriocarcinoma by Mediating TGF-β1 Expression
Hanxi Yu1, Weichen Zhang2, Peilin Han1
1Department of Gynecology, The First Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou 310006, People's Republic of China.
Background:
Mammalian Ste20-like kinase 4 (MST4), also known as serine/threonine kinase 26 (STK26), promotes development of several cancers and is found to be highly expressed in the placenta. However, in choriocarcinoma that originated from the placenta, the expression of MST4 was undetermined and its mechanism was unknown. In this study, the expression of MST4 in choriocarcinoma as well as the underlying mechanism was explored.
Purpose:
To detect the expression of MST4 in patient samples and mechanism of mediating EMT by MST4 in choriocarcinoma.
Patients And Methods:
The metastatic lesions of choriocarcinoma (n=17) and volunteer villus (n=17) were collected to determine MST4 expression using immunohistochemistry and H&E staining. We use siRNA and lentiviral vector to knockdown MST4 and use plasmid to overexpress MST4 in choriocarcinoma. Then, we apply real-time polymerase chain reaction (RT-PCR), Western blot assay and immunofluorescence assay to detect target protein expressions. Cell invasion and migration and cell proliferation were detected by transwell assay and wound healing assay and CCK-8 and cell colony formation.
Results:
MST4 is lowly expressed in the metastatic lesions of choriocarcinoma patients when compared with normal villus. Knockdown of MST4 activated epithelial-mesenchymal transition (EMT) process, significantly increasing the ability of invasion and migration in choriocarcinoma cell lines (JAR and JEG-3). In contrast, the EMT process was restrained in choriocarcinoma cell lines with overexpressed MST4. Meanwhile, genome-wide gene expression array, Western blot and ELISA revealed that tumor growth factor-beta 1 (TGF-β1) has significantly increased. The EMT process and metastatic prompting biofunction were reversed after using TGF-β1 inhibitor (LY364947) in the choriocarcinoma cell lines with MST4 knockdown.
Conclusion:
Our studies demonstrated that MST4 was lowly expressed in patient samples. Additionally, JAR and JEG-3 increase cell invasion and migration ability while there is no influence on cell proliferation with MST4 knockdown. Conversely, the metastatic ability of JAR and JEG-3 was decreased with overexpressed MST4. Moreover, TGF-β1 was a key factor after MST4 knockdown. In conclusion, MST4 affects choriocarcinoma EMT by mediating TGF-β1 expression.
Insights
Mammalian Ste20-like kinase 4 (MST4) is lowly expressed in choriocarcinoma, promoting cancer cell invasion and migration by mediating TGF-β1. Overexpressing MST4 restrains metastasis, suggesting MST4 as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Mammalian Ste20-like kinase 4 (MST4), also known as serine/threonine kinase 26 (STK26), is implicated in cancer development and highly expressed in the placenta.
- The expression and mechanism of MST4 in choriocarcinoma, a cancer originating from placental tissue, remained largely undetermined.
- This study investigates MST4 expression and its role in choriocarcinoma pathogenesis.
Purpose of the Study:
- To determine MST4 expression levels in choriocarcinoma patient samples.
- To elucidate the mechanism by which MST4 mediates epithelial-mesenchymal transition (EMT) in choriocarcinoma.
- To assess the impact of MST4 modulation on choriocarcinoma cell behavior.
Main Methods:
- Immunohistochemistry and H&E staining were used to analyze MST4 expression in choriocarcinoma tissues and normal villi.
- In vitro studies involved siRNA-mediated MST4 knockdown and lentiviral/plasmid-based overexpression in choriocarcinoma cell lines (JAR and JEG-3).
- RT-PCR, Western blot, immunofluorescence, transwell assays, wound healing assays, CCK-8, and colony formation assays were employed to evaluate gene/protein expression and cellular functions.
Main Results:
- MST4 expression was significantly lower in metastatic choriocarcinoma lesions compared to normal villi.
- MST4 knockdown enhanced choriocarcinoma cell invasion and migration by activating EMT, while MST4 overexpression suppressed these metastatic properties.
- Tumor growth factor-beta 1 (TGF-β1) expression increased upon MST4 knockdown, and its inhibition reversed the pro-metastatic effects.
Conclusions:
- MST4 is downregulated in choriocarcinoma, and its reduction promotes cell invasion and migration, independent of proliferation.
- MST4 overexpression inhibits the metastatic potential of choriocarcinoma cells.
- MST4 influences choriocarcinoma EMT by regulating TGF-β1 expression, highlighting MST4 as a potential therapeutic target.
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