Role of sialylation in 3D multicellular tumor spheroid formation and in vivo tumors

High-grade serous ovarian cancer (HGSOC) can originate in the fallopian tube and then spread to the ovary. Our objective was to evaluate the role of multicellular tumor spheroids (MTS) in ovarian metastasis. By testing a panel of murine oviductal epithelial (MOE) cells with genetic alterations mimicking those seen in HGSOC, we found that loss of PTEN allowed MTS formation under ultra-low adhesion conditions. Confirming these results in vivo, MTS-like structures were observed in the oviducts of PAX8Cre/+ PTENflox/flox mice. MOE PTENshRNA cells could incorporate up to 25% wild type cells into MTS, while higher percentages of wild type cells resulted in a loss of MTS formation. MTS formation allowed MOE PTENshRNA cells to survive better under ultra-low adhesion conditions than control cells. MTS also attached to the ovarian stroma, as would be exposed during ovulation. Interestingly, MTS more robustly cleared monolayers of murine ovarian surface epithelia than murine ovarian fibroblasts. When xenografted into the ovarian bursa, OVCAR8 MTS were able to form tumors in the ovary at a similar rate as an equal number of OVCAR8 cells grown on traditional cell culture plastic. In conclusion, loss of a single gene (PTEN) allows the fallopian tube epithelia to form MTS, which survive better under ultra-low adhesion conditions, attach to the extracellular matrix exposed during ovulation, and colonize the ovary. These results suggest that MTS may contribute to seeding of the ovary in HGSOC patients.

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Blocking the GITR-GITRL pathway to overcome resistance to therapy in sarcomatoid malignant pleural mesothelioma

Communications Biology ◽

10.1038/s42003-021-02430-5 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Meilin Chan ◽

Licun Wu ◽

Zhihong Yun ◽

Trevor D. McKee ◽

Michael Cabanero ◽

...

Keyword(s):

Tumor Growth ◽

Malignant Pleural Mesothelioma ◽

Neutralizing Antibodies ◽

Pleural Mesothelioma ◽

Spheroid Formation ◽

Resistance To Therapy ◽

Sarcomatoid Mesothelioma

AbstractMalignant pleural mesothelioma (MPM) is an aggressive neoplasm originating from the pleura. Non-epithelioid (biphasic and sarcomatoid) MPM are particularly resistant to therapy. We investigated the role of the GITR-GITRL pathway in mediating the resistance to therapy. We found that GITR and GITRL expressions were higher in the sarcomatoid cell line (CRL5946) than in non-sarcomatoid cell lines (CRL5915 and CRL5820), and that cisplatin and Cs-137 irradiation increased GITR and GITRL expressions on tumor cells. Transcriptome analysis demonstrated that the GITR-GITRL pathway was promoting tumor growth and inhibiting cell apoptosis. Furthermore, GITR+ and GITRL+ cells demonstrated increased spheroid formation in vitro and in vivo. Using patient derived xenografts (PDXs), we demonstrated that anti-GITR neutralizing antibodies attenuated tumor growth in sarcomatoid PDX mice. Tumor immunostaining demonstrated higher levels of GITR and GITRL expressions in non-epithelioid compared to epithelioid tumors. Among 73 patients uniformly treated with accelerated radiation therapy followed by surgery, the intensity of GITR expression after radiation negatively correlated with survival in non-epithelioid MPM patients. In conclusion, the GITR-GITRL pathway is an important mechanism of autocrine proliferation in sarcomatoid mesothelioma, associated with tumor stemness and resistance to therapy. Blocking the GITR-GITRL pathway could be a new therapeutic target for non-epithelioid mesothelioma.

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Multicellular tumor spheroid formation in duplex microcapsules for analysis of chemosensitivity

Cancer Science ◽

10.1111/j.1349-7006.2011.02187.x ◽

2012 ◽

Vol 103 (3) ◽

pp. 549-554 ◽

Cited By ~ 16

Author(s):

Shinji Sakai ◽

Kazuya Inamoto ◽

Yang Liu ◽

Shinji Tanaka ◽

Shigeki Arii ◽

...

Keyword(s):

Tumor Spheroid ◽

Spheroid Formation ◽

Multicellular Tumor Spheroid

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A multicellular tumor spheroid formation and extraction chip

10th IEEE International Conference on Nanotechnology ◽

10.1109/nano.2010.5697838 ◽

2010 ◽

Author(s):

Hye-Jin Jin ◽

Taeyoon Kim ◽

Young-Ho Cho ◽

Jin-Mo Gu ◽

Jhingook Kim ◽

...

Keyword(s):

Tumor Spheroid ◽

Spheroid Formation ◽

Multicellular Tumor Spheroid

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In vitro lung cancer multicellular tumor spheroid formation using a microfluidic device

Biotechnology and Bioengineering ◽

10.1002/bit.27114 ◽

2019 ◽

Vol 116 (11) ◽

pp. 3041-3052 ◽

Cited By ~ 8

Author(s):

Sang Woo Lee ◽

Soyoung Hong ◽

Boyoung Jung ◽

Soo Yeon Jeong ◽

Jae Hee Byeon ◽

...

Keyword(s):

Lung Cancer ◽

Microfluidic Device ◽

Tumor Spheroid ◽

Spheroid Formation ◽

Multicellular Tumor Spheroid

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CHI3L1 results in poor outcome of ovarian cancer by promoting properties of stem-like cells

Endocrine Related Cancer ◽

10.1530/erc-18-0300 ◽

2019 ◽

Vol 26 (1) ◽

pp. 73-88 ◽

Cited By ~ 3

Author(s):

Han-Wei Lin ◽

Ying-Cheng Chiang ◽

Nai-Yun Sun ◽

Yu-Li Chen ◽

Chi-Fang Chang ◽

...

Keyword(s):

Ovarian Cancer ◽

Cancer Cells ◽

Signaling Pathways ◽

Cancer Patients ◽

Side Population ◽

Ovarian Cancer Cells ◽

Tumor Spheroid ◽

Spheroid Formation ◽

Poor Outcome

The role of chitinase-3-like protein 1 (CHI3L1) in ovarian cancer and the possible mechanisms were elucidated. CHI3L1 is a secreted glycoprotein and associated with inflammation, fibrosis, asthma, extracellular tissue remodeling and solid tumors. Our previous study showed CHI3L1 could be a potential prognostic biomarker for epithelial ovarian cancer and could protect cancer cells from apoptosis. Therefore, clinical data and quantitation of CHI3L1 of ovarian cancer patients, tumor spheroid formation, side-population assays, Aldefluor and apoptotic assays, ELISA, RT-PCR, immunoblotting and animal experiments were performed in two ovarian cancer cells lines, OVCAR3 and CA5171, and their CHI3L1-overexpressing and -knockdown transfectants. High expression of CHI3L1 was associated with poor outcome and chemoresistance in ovarian cancer patients. The mRNA expression of CHI3L1 in CA5171 ovarian cancer stem-like cells was 3-fold higher than in CA5171 parental cells. CHI3L1 promoted the properties of ovarian cancer stem-like cells including generating more and larger tumor spheroids and a higher percentage of ALDH+ in tumor cells and promoting resistance to cytotoxic drug-induced apoptosis. CHI3L1 could induce both the Akt (essential) and Erk signaling pathways, and then enhance expression of β-catenin followed by SOX2, and finally promote tumor spheroid formation and other properties of ovarian cancer stem-like cells. OVCAR3 CHI3L1-overexpressing transfectants were more tumorigenic in vivo, whereas CA5171 CHI3L1-knockdown transfectants were not tumorigenic in vivo. CHI3L1 critically enhances the properties of ovarian cancer stem-like cells. CHI3L1 or CHI3L1-regulated signaling pathways and molecules could be potential therapeutic targets in ovarian cancer.

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