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MYOCD and SMAD3/SMAD4 form a positive feedback loop and drive TGF-β-induced epithelial–mesenchymal transition in non-small cell lung cancer

Oncogene volume 39, pages 2890–2904 (2020)Cite this article

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Abstract

Myocardin (MYOCD) promotes Smad3-mediated transforming growth factor-β (TGF-β) signaling in mouse fibroblast cells. Our previous studies show that TGF-β/SMADs signaling activation enhances epithelial–mesenchymal transition (EMT) in human non-small cell lung cancer (NSCLC) cells. However, whether and how MYOCD contributes to TGF-β-induced EMT of NSCLC cells are poorly elucidated. Here, we found that TGF-β-induced EMT was accompanied by increased MYOCD expression. Interestingly, MYOCD overexpression augmented EMT and invasion of NSCLC cells induced by TGF-β, whereas knockdown of MYOCD expression attenuated these effects. Overexpression and knockdown of MYOCD resulted in the upregulation and downregulation of TGF-β-induced Snail mRNA, respectively. Moreover, MYOCD overexpression promoted TGF-β-stimulated NSCLC cell metastasis in vivo. MYOCD was highly expressed and positively correlated with Snail in metastatic NSCLC tissues. Mechanistically, MYOCD directly interacted with SMAD3 and sustained the formation of TGF-β-induced nuclear SMAD3/SMAD4 complex, facilitating TGF-β/SMAD3-induced transactivation of Snail. Importantly, MYOCD was transcriptionally activated by TGF-β-induced SMAD3/SMAD4 complex and CRISPR/Cas9-mediated silencing of SMAD3/SMAD4 led to a reduction in MYOCD mRNA expression. Taken together, our findings indicate that MYOCD promotes TGF-β-induced EMT and metastasis of NSCLC and identify a positive feedback loop between MYOCD and SMAD3/SMAD4 driving TGF-β-induced EMT.

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Fig. 1: MYOCD expression is increased in TGF-β-induced EMT of NSCLC cells.
Fig. 2: MYOCD is transcriptionally activated by TGF-β-induced SMAD3/4 complex in NSCLC cells.
Fig. 3: MYOCD enhances the sustained activation of TGF-β-induced nuclear SMAD3/SMAD4 complex.
Fig. 4: MYOCD promotes TGF-β/SMAD3-induced transcriptional activity by interacting with SMAD3.
Fig. 5: MYOCD knockdown impairs TGF-β1-activated recruitment of p-SMAD3 to the Snail promoter, and Snail knockdown inhibits TGF-β1-induced migration and invasion of A549 cells.
Fig. 6: MYOCD overexpression boosts TGF-β-induced EMT and invasion of NSCLC cells.
Fig. 7: MYOCD overexpression promotes TGF-β-stimulated NSCLC cell metastasis in vivo.
Fig. 8: MYOCD expression is positively correlated with Snail expression in metastatic NSCLC tissues.

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Acknowledgements

We are grateful for participation and cooperation from the patients with NSCLC. Funding was provided by grants from National Natural Science Foundation of China (81872343, 81672277), and Suzhou Key Laboratory for Molecular Cancer Genetics (SZS201209), and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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  1. These authors contributed equally: Xin Tong, Shengjie Wang, Zhe Lei, Chang Li

Authors and Affiliations

  1. Soochow University Laboratory of Cancer Molecular Genetics, Medical College of Soochow University, Suzhou, 215123, Jiangsu, China

    Xin Tong, Shengjie Wang, Zhe Lei, Cuijuan Zhang, Zhiyue Su, Xia Liu & Hong-Tao Zhang

  2. Department of Genetics, School of Biology and Basic Medical Sciences, Medical College of Soochow University, Suzhou, 215123, Jiangsu, China

    Xin Tong, Shengjie Wang, Zhe Lei, Cuijuan Zhang, Zhiyue Su, Xia Liu & Hong-Tao Zhang

  3. Department of Basic Medicine, Kangda College of Nanjing Medical University, Lianyungang, 222000, China

    Shengjie Wang

  4. Department of Thoracic Surgery, The First Affiliated Hospital of Soochow University, Medical College of Soochow University, Suzhou, 215006, Jiangsu, China

    Chang Li & Jun Zhao

  5. Suzhou Key Laboratory for Molecular Cancer Genetics, Suzhou, 215123, Jiangsu, China

    Hong-Tao Zhang

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Contributions

Conception and design: XT and H-TZ. Development of methodology: XT, SW, and ZL. Acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.): XT, SW, ZL, CL, CZ, ZS, XL, and JZ. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, computational analysis): XT, SW, ZL, CL, and CZ. Writing, review, and/or revision of the manuscript: XT and H-TZ. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): XT, SW, ZL, CL, and CZ. Study supervision: H-TZ.

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Tong, X., Wang, S., Lei, Z. et al. MYOCD and SMAD3/SMAD4 form a positive feedback loop and drive TGF-β-induced epithelial–mesenchymal transition in non-small cell lung cancer. Oncogene 39, 2890–2904 (2020). https://doi.org/10.1038/s41388-020-1189-4

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