Assessment of Ovarian Cancer Spheroid Attachment and Invasion of Mesothelial Cells in Real Time

Abstract Background Metastasis is the key cause of death in ovarian cancer patients. To figure out the biological nature of cancer metastasis is essential for developing effective targeted therapy. Here we investigate how long non-coding RNA (lncRNA) SPOCD1-AS from ovarian cancer extracellular vesicles (EVs) remodel mesothelial cells through a mesothelial-to-mesenchymal transition (MMT) manner and facilitate peritoneal metastasis. Methods EVs purified from ovarian cancer cells and ascites of patients were applied to mesothelial cells. The MMT process of mesothelial cells was assessed by morphology observation, western blot analysis, migration assay and adhesion assay. Altered lncRNAs of EV-treated mesothelial cells were screened by RNA sequencing and identified by qRT-PCR. SPOCD1-AS was overexpressed or silenced by overexpression lentivirus or shRNA, respectively. RNA pull-down and RNA immunoprecipitation assays were conducted to reveal the mechanism by which SPOCD1-AS remodeled mesothelial cells. Interfering peptides were synthesized and applied. Ovarian cancer orthotopic implantation mouse model was established in vivo. Results We found that ovarian cancer-secreted EVs could be taken into recipient mesothelial cells, induce the MMT phenotype and enhance cancer cell adhesion to mesothelial cells. Furthermore, SPOCD1-AS embedded in ovarian cancer-secreted EVs was transmitted to mesothelial cells to induce the MMT process and facilitate peritoneal colonization in vitro and in vivo. SPOCD1-AS induced the MMT process of mesothelial cells via interacting with G3BP1 protein. Additionally, G3BP1 interfering peptide based on the F380/F382 residues was able to block SPOCD1-AS/G3BP1 interaction, inhibit the MMT phenotype of mesothelial cells, and diminish peritoneal metastasis in vivo. Conclusions Our findings elucidate the mechanism associated with EVs and their cargos in ovarian cancer peritoneal metastasis and may provide a potential approach for metastatic ovarian cancer therapeutics.

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Clinical Utility of Real-Time Targeted Molecular Profiling in the Clinical Management of Ovarian Cancer: The ALLOCATE Study

JCO Precision Oncology ◽

10.1200/po.19.00019 ◽

2019 ◽

pp. 1-18

Author(s):

Olga Kondrashova ◽

Gwo-Yaw Ho ◽

George Au-Yeung ◽

Leakhena Leas ◽

Tiffany Boughtwood ◽

...

Keyword(s):

Ovarian Cancer ◽

Real Time ◽

Targeted Therapies ◽

Clinical Management ◽

Clinical Utility ◽

Advanced Ovarian Cancer ◽

Molecular Profiling ◽

Molecular Testing ◽

Low Grade ◽

Ovarian Carcinomas

PURPOSE The ALLOCATE study was designed as a pilot to demonstrate the feasibility and clinical utility of real-time targeted molecular profiling of patients with recurrent or advanced ovarian cancer for identification of potential targeted therapies. PATIENTS AND METHODS A total of 113 patients with ovarian cancer of varying histologies were recruited from two tertiary hospitals, with 99 patient cases suitable for prospective analysis. Targeted molecular and methylation profiling of fresh biopsy and archived tumor samples were performed by screening for mutations or copy-number variations in 44 genes and for promoter methylation of BRCA1 and RAD51C. RESULTS Somatic genomic or methylation events were identified in 85% of all patient cases, with potentially actionable events with defined targeted therapies (including four resistance events) detected in 60% of all patient cases. On the basis of these findings, six patients received molecularly guided therapy, three patients had unsuspected germline cancer–associated BRCA1/ 2 mutations and were referred for genetic counseling, and two intermediate differentiated (grade 2) serous ovarian carcinomas were reclassified as low grade, leading to changes in clinical management. Additionally, secondary reversion mutations in BRCA1/ 2 were identified in fresh biopsy samples of two patients, consistent with clinical platinum/poly (ADP-ribose) polymerase inhibitor resistance. Timely reporting of results if molecular testing is done at disease recurrence, as well as early referral for patients with platinum-resistant cancers, were identified as factors that could improve the clinical utility of molecular profiling. CONCLUSION ALLOCATE molecular profiling identified known genomic and methylation alterations of the different ovarian cancer subtypes and was deemed feasible and useful in routine clinical practice. Better patient selection and access to a wider range of targeted therapies or clinical trials will further enhance the clinical utility of molecular profiling.

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Senescent Peritoneal Mesothelial Cells Promote Ovarian Cancer Cell Adhesion

American Journal Of Pathology ◽

10.2353/ajpath.2009.080613 ◽

2009 ◽

Vol 174 (4) ◽

pp. 1230-1240 ◽

Cited By ~ 43

Author(s):

Krzysztof Ksiazek ◽

Justyna Mikula-Pietrasik ◽

Katarzyna Korybalska ◽

Grzegorz Dworacki ◽

Achim Jörres ◽

...

Keyword(s):

Ovarian Cancer ◽

Cell Adhesion ◽

Cancer Cell ◽

Ovarian Cancer Cell ◽

Mesothelial Cells ◽

Peritoneal Mesothelial Cells ◽

Cancer Cell Adhesion

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Abstract AP10: REAL-TIME ASSESSMENT OF HGSC DNA DAMAGE REPAIR DEFECTS AND DEFECT-INDUCED RESPONSE TO THERAPY IN OVARIAN CANCER ORGANOIDS

10.1158/1557-3265.ovcasymp18-ap10 ◽

2019 ◽

Author(s):

Sarah J. Hill ◽

Brennan Decker ◽

Emma A. Roberts ◽

Chunyu Yang ◽

Neil S. Horowitz ◽

...

Keyword(s):

Ovarian Cancer ◽

Dna Damage ◽

Real Time ◽

Dna Damage Repair ◽

Response To Therapy ◽

Induced Response ◽

Damage Repair ◽

Time Assessment

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Multicolorin vivotargeted imaging to guide real-time surgery of HER2-positive micrometastases in a two-tumor coincident model of ovarian cancer

Cancer Science ◽

10.1111/j.1349-7006.2009.01133.x ◽

2009 ◽

Vol 100 (6) ◽

pp. 1099-1104 ◽

Cited By ~ 38

Author(s):

Michelle Longmire ◽

Nobuyuki Kosaka ◽

Mikako Ogawa ◽

Peter L. Choyke ◽

Hisataka Kobayashi

Keyword(s):

Ovarian Cancer ◽

Real Time ◽

Her2 Positive

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Increased Expression of the Pro-Protein Convertase Furin Predicts Decreased Survival in Ovarian Cancer

Analytical Cellular Pathology ◽

10.1155/2007/930321 ◽

2007 ◽

Vol 29 (4) ◽

pp. 289-299

Author(s):

Robert E. Page ◽

Andrés J. P. Klein-Szanto ◽

Samuel Litwin ◽

Emmanuelle Nicolas ◽

Raid Al-Jumaily ◽

...

Keyword(s):

Ovarian Cancer ◽

Tumor Progression ◽

Real Time ◽

Cell Lines ◽

Cancer Cell ◽

Real Time Pcr ◽

Ovarian Tumor ◽

Cancer Cell Lines ◽

Rt Pcr ◽

Primary Tumors

Background: Proprotein convertases (PCs) are serine proteases that after restricted proteolysis activate many proteins that play a crucial role in cancer such as metalloproteinases, growth factors and growth factor receptors, adhesion molecules, and angiogenic factors. Although the expression of several PCs is increased in many tumors, their expression in primary ovarian tumors has not been studied in detail. We sought to determine if there was an association between the expression of the ubiquitously expressed PCs, furin, PACE-4, PC-5 and PC-7, and ovarian tumor progression. Methods: We assessed their expression by RT-PCR, Real-time PCR, Western blot, and immunohistochemistry using cells derived from normal human ovarian surface epithelium (HOSE) and cancer cell lines as well as ovarian epithelial cancer specimens (45 RT-PCR/Real-time PCR, and 120 archival specimens for Immunohistochemistry). Results: We found that furin expression was restricted to the cancer cell lines. In contrast, PACE-4 and PC-7 showed expression only in normal HOSE cells lines. Furthermore, furin was predominantly expressed in primary tumors from patients who survived for less than five years. The other PCs are either expressed in the group of survivors (PC-7 and PACE4) or expressed in low amounts (PC-5). Conclusions: Our studies point to a clear relationship between furin and ovarian cancer. In addition, these results show that furin exhibits the closest association with ovarian cancer among the ubiquitously expressed PCs, arguing against the redundancy of these proteases. In summary, furin may constitute a marker for ovarian tumor progression and could contribute to predict the outcome of this disease.

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Monitoring of ovarian cancer cell invasion in real time with frequency-dependent impedance measurement

AJP Cell Physiology ◽

10.1152/ajpcell.00211.2016 ◽

2016 ◽

Vol 311 (6) ◽

pp. C1040-C1047 ◽

Cited By ~ 4

Author(s):

Chun-Min Lo ◽

Jun-Chih Lo ◽

Priscila Y. Sato ◽

Tsz-Lun Yeung ◽

Samuel C. Mok ◽

...

Keyword(s):

Ovarian Cancer ◽

Cancer Cells ◽

Real Time ◽

Cell Invasion ◽

Umbilical Vein ◽

Ovarian Cancer Cells ◽

Adherent Cells ◽

Frequency Dependent ◽

Average Cell ◽

Cell Substrate

The conventional approach to assessing cancer invasion is primarily for end-point analysis, which does not provide temporal information on the invasion process or any information on the interactions between invading cells and the underlying adherent cells. To alleviate these limitations, the present study exploited electric cell-substrate impedance sensing (ECIS) to monitor the invasion of ovarian cancer cells (SKOV-3) through an adherent monolayer of human umbilical vein endothelial cells (HUVECs). Impedance was measured at 4 kHz of AC voltage or was measured as a function of AC frequency (25 Hz to 60 kHz). By measuring impedance at 4-kHz AC, we found that the invasion of SKOV-3 cells through the HUVEC monolayer was manifested as a rapid decrease in transendothelial electrical resistance in real time. The invasion was augmented in the presence of hepatocyte growth factor (HGF). The enhancing effect of HGF was attenuated by c-Met inhibitor (SU11274). By measuring the frequency-dependent impedance of SKOV-3 cells over time, we found that HGF-enhanced SKOV-3 cell invasion was accomplished with reduced junctional resistance ( Rb), increased average cell-substrate separation ( h), and increased micromotion. SU11274 attenuated the effects of HGF on Rb, h, and micromotion in the SKOV-3 monolayer. SU11274 also increased the barrier function of the HUVEC monolayer by increasing Rb and decreasing h. In conclusion, this study demonstrated an improved method for monitoring and studying the interactions between cancer cells and the underlying adherent cells during invasion in real time. Alterations in cellular biophysical properties ( Rb, h) associated with cancer transendothelial invasion were detected.

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