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MicroRNA-200 family governs ovarian inclusion cyst formation and mode of ovarian cancer spread

Abstract

Epidemiologic and histopathologic findings and the laying hen model support the long-standing incessant ovulation hypothesis and cortical inclusion cyst involvement in sporadic ovarian cancer development. MicroRNA-200 (miR-200) family is highly expressed in ovarian cancer. Herewith, we show that ovarian surface epithelial (OSE) cells with ectopic miR-200 expression formed stabilized cysts in three-dimensional (3D) organotypic culture with E-cadherin fragment expression and steroid hormone pathway activation, whereas ovarian cancer 3D cultures with miR-200 knockdown showed elevated TGF-β expression, mitotic spindle disorientation, increased lumenization, disruption of ROCK-mediated myosin II phosphorylation, and SRC signaling, which led to histotype-dependent loss of collective movement in tumor spread. Gene expression profiling revealed that epithelial–mesenchymal transition and hypoxia were the top enriched gene sets regulated by miR-200 in both OSE and ovarian cancer cells. The molecular changes uncovered by the in vitro studies were verified in both human and laying hen ovarian cysts and tumor specimens. As miR-200 is also essential for ovulation, our results of estrogen pathway activation in miR-200-expressing OSE cells add another intriguing link between incessant ovulation and ovarian carcinogenesis.

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Fig. 1: Ovarian cell lines with aberrant miR-200 expression showed reduced growth rates and changes in marker expression.
Fig. 2: Overexpression of miR-200 stabilized OSE7 spheroids.
Fig. 3: Elevated expression of miR-200 in human and laying hen inclusion cysts and ovarian tumor tissues and association with estrogen response protein TFF1.
Fig. 4: Ovarian cancer spheroids with miR-200 KD had increased number of lumina and defects in mitotic spindle orientation.
Fig. 5: TGF-β regulated myosin II phosphorylation for the single-cell migration in ovarian cancer cells with miR-200 knockdown.
Fig. 6: miR-200-regulated SRC phosphorylation for cell motility and tumor spread.

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Acknowledgements

We acknowledge Dr. Gregory Goodall for the provision of lentiviral miR-200 expression constructs and control, Mr. Shi-Kin Ma for the help of constructing the polar graphs, the support of the CNOOC Grants to the first author, and the Robert and Deborah First Fund, the Sperling Family Fund Foundation, Ruth N. White Gynecologic Oncology Research Fund, Women’s Cancer Program and Gillette Center for Women’s Cancer from Dana–Farber Cancer Institute, Ovarian Cancer Research Foundation, Adler Foundation, Inc., and Friends of Dana–Farber Cancer Institute to The Laboratory of Gynecologic Oncology at Brigham and Women’s Hospital.

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Correspondence to Wing-Ping Fong.

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Choi, PW., So, W.W., Yang, J. et al. MicroRNA-200 family governs ovarian inclusion cyst formation and mode of ovarian cancer spread. Oncogene 39, 4045–4060 (2020). https://doi.org/10.1038/s41388-020-1264-x

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