Cannabidiol: Assessing preclinical safety in ovarian and endometrial carcinoma cell lines.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e24130-e24130
Author(s):  
Shannon Rush ◽  
Arvinder K Kapur ◽  
Manish S. Patankar ◽  
Lisa Marie Barroilhet
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Symptomatic Relief ◽  
Preclinical Research ◽  
Cancer Symptoms ◽  
Carcinoma Cell Lines ◽  
Chemotherapy Side Effects ◽  
Cell Pellet ◽  
X 24 ◽  
Cayman Chemical

e24130 Background: Cancer patients use cannabidiol (CBD) for chemotherapy and cancer symptoms, though research of CBD safety and efficacy for these conditions are ongoing and mixed. We sought to determine endometrial (ECC1) and epithelial ovarian cancer (Kuramochi) cell proliferation when exposed to different concentrations of CBD, for the broader goal to establish if CBD can safely be utilized to treat the symptoms of cancer, including those caused by chemotherapy. Methods: ECC1 and Kuramochi cells were kept in media (RPMI with 10% bovine serum and 1% penicillin/streptomycin). We passaged cells when > 90% confluent by adding tryspin-EDTA, incubating at 37C for 3 minutes, then spinning down with media to harvest the cell pellet. Cells were re-suspended in media, counted and apportioned to 96 well plates. Plates were incubated at 37C x 24 hours. CBD (from Cayman Chemical) was suspended in DMSO per manufacturer instruction then used to treat cells x72 hours at different concentrations (2.5-50uM). MTT was added to cells, cells incubated at 37C x 3 hours, media and MTT were removed and DMSO was added. Optical depth (OD) was calculated for plates using SoftMaxPro version 6.2.2. ODs were used to calculate inhibitory concentration for 50% cell death (IC50). Results: Kuramochi and ECC1 demonstrated decreased cell proliferation when exposed to CBD for 72hours. ECC1 IC50 fell between 2.5-5uM. Kuramochi IC50 fell between 15-20uM. Nearly all ECC1 growth was inhibited at concentrations 10uM or greater. Kuramochi proliferation was 15% that of controls at concentrations of 40 and 50uM CBD. Conclusions: ECC1 and Kuramochi cells demonstrated decreased proliferation in the presence of CBD. This bodes well for future studies of concurrent exposure to CBD and cytotoxic chemotherapy. Further preclinical research needed on CBD effects in endometrial and ovarian cancer, as patients turn to CBD for symptomatic relief from cancer and chemotherapy side effects. [Table: see text]

scholarly journals Wnt5A modulates integrin expression in a receptor-dependent manner in ovarian cancer cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Vajihe Azimian-Zavareh ◽  
Zeinab Dehghani-Ghobadi ◽  
Marzieh Ebrahimi ◽  
Kian Mirzazadeh ◽  
Irina Nazarenko ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Cell Attachment ◽  
Ovarian Cancer Cells ◽  
Integrin Expression ◽  
Dependent Manner ◽  
Cancer Cell Proliferation ◽  
Expression Levels ◽  
Multicellular Aggregates ◽  
Focal Adhesion Kinase Activity

AbstractWnt5A signals through various receptors that confer versatile biological functions. Here, we used Wnt5A overexpressing human ovarian SKOV-3 and OVCAR-3 stable clones for assessing integrin expression, cell proliferation, migration, invasion, and the ability of multicellular aggregates (MCAs) formation. We found here, that Wnt5A regulates differently the expression of its receptors in the stable Wnt5A overexpressing clones. The expression levels of Frizzled (FZD)-2 and -5, were increased in different clones. However ROR-1, -2 expression levels were differently regulated in clones. Wnt5A overexpressing clones showed increased cell proliferation, migration, and clonogenicity. Moreover, Wnt5A overexpressing SKOV-3 clone showed increased MCAs formation ability. Cell invasion had been increased in OVCAR-3-derived clones, while this was decreased in SKOV-3-derived clone. Importantly, αv integrin expression levels were increased in all assessed clones, accompanied by increased cell attachment to fibronectin and focal adhesion kinase activity. Moreover, the treatment of clones with Box5 as a Wnt5A/FZD5 antagonist abrogates ITGAV increase, cell proliferation, migration, and their attachment to fibronectin. Accordingly, we observed significantly higher expression levels of ITGAV and ITGB3 in human high-grade serous ovarian cancer specimens and ITGAV correlated positively with Wnt5A in metastatic serous type ovarian cancer. In summary, we hypothesize here, that Wnt5A/FZD-5 signaling modulate αv integrin expression levels that could be associated with ovarian cancer cell proliferation, migration, and fibronectin attachment.

scholarly journals Structural Bases for the Synergistic Inhibition of Human Thymidylate Synthase and Ovarian Cancer Cell Growth by Drug Combinations

Cancers ◽  
2021 ◽  
Vol 13 (9) ◽  
pp. 2061
Author(s):  
Cecilia Pozzi ◽  
Matteo Santucci ◽  
Gaetano Marverti ◽  
Domenico D’Arca ◽  
Lorenzo Tagliazucchi ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Growth ◽  
Cancer Cell ◽  
Thymidylate Synthase ◽  
Ovarian Cancer Cell ◽  
Ovarian Cancer Cells ◽  
Preclinical Research ◽  
Cancer Cell Growth ◽  
X Ray ◽  
Synergistic Inhibition

Combining drugs represent an approach to efficiently prevent and overcome drug resistance and to reduce toxicity; yet it is a highly challenging task, particularly if combinations of inhibitors of the same enzyme target are considered. To show that crystallographic and inhibition kinetic information can provide indicators of cancer cell growth inhibition by combinations of two anti-human thymidylate synthase (hTS) drugs, we obtained the X-ray crystal structure of the hTS:raltitrexed:5-fluorodeoxyuridine monophosphate (FdUMP) complex. Its analysis showed a ternary complex with both molecules strongly bound inside the enzyme catalytic cavity. The synergistic inhibition of hTS and its mechanistic rationale were consistent with the structural analysis. When administered in combination to A2780 and A2780/CP ovarian cancer cells, the two drugs inhibited ovarian cancer cell growth additively/synergistically. Together, these results support the idea that X-ray crystallography can provide structural indicators for designing combinations of hTS (or any other target)-directed drugs to accelerate preclinical research for therapeutic application.

scholarly journals The Epithelial–Mesenchymal Transcription Factor SNAI1 Represses Transcription of the Tumor Suppressor miRNA let-7 in Cancer

Cancers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1469
Author(s):  
Hanmin Wang ◽  
Evgeny Chirshev ◽  
Nozomi Hojo ◽  
Tise Suzuki ◽  
Antonella Bertucci ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Stem Cell ◽  
Cancer Cells ◽  
Epithelial Mesenchymal Transition ◽  
Tumor Burden ◽  
Cell Phenotype ◽  
Mesenchymal Transition ◽  
Carcinoma Cell Lines ◽  
Future Work

We aimed to determine the mechanism of epithelial–mesenchymal transition (EMT)-induced stemness in cancer cells. Cancer relapse and metastasis are caused by rare stem-like cells within tumors. Studies of stem cell reprogramming have linked let-7 repression and acquisition of stemness with the EMT factor, SNAI1. The mechanisms for the loss of let-7 in cancer cells are incompletely understood. In four carcinoma cell lines from breast cancer, pancreatic cancer, and ovarian cancer and in ovarian cancer patient-derived cells, we analyzed stem cell phenotype and tumor growth via mRNA, miRNA, and protein expression, spheroid formation, and growth in patient-derived xenografts. We show that treatment with EMT-promoting growth factors or SNAI1 overexpression increased stemness and reduced let-7 expression, while SNAI1 knockdown reduced stemness and restored let-7 expression. Rescue experiments demonstrate that the pro-stemness effects of SNAI1 are mediated via let-7. In vivo, nanoparticle-delivered siRNA successfully knocked down SNAI1 in orthotopic patient-derived xenografts, accompanied by reduced stemness and increased let-7 expression, and reduced tumor burden. Chromatin immunoprecipitation demonstrated that SNAI1 binds the promoters of various let-7 family members, and luciferase assays revealed that SNAI1 represses let-7 transcription. In conclusion, the SNAI1/let-7 axis is an important component of stemness pathways in cancer cells, and this study provides a rationale for future work examining this axis as a potential target for cancer stem cell-specific therapies.

Lysine-specific demethylase 1 (LSD1) promotes ovarian cancer cell progression by Forkhead box O 3a (FOXO3a) inhibition

2020 ◽  
Vol 10 (4) ◽  
pp. 594-602 ◽  
Author(s):  
Li Liu ◽  
Fuxing Hao ◽  
Anping Wang ◽  
Xiaolan Chen ◽  
Bin Zhang ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cell Proliferation ◽  
Cancer Cell ◽  
Ovarian Cancer Cell ◽  
Soft Agar ◽  
Ovarian Cancer Cells ◽  
Cancer Cell Growth ◽  
Forkhead Box ◽  
Lysine Specific Demethylase ◽  
Cell Progression

Recently, LSD1 is considered as a possible therapeutic mark for ovarian epithelial cancer (OEC). Though, the underlying molecular mechanism by which LSD1 endorses the oncogenesis of OEC has not been fully understood. Here, we revealed that overexpression of LSD1 downregulated Forkhead box O 3a (FOXO3a), while knockdown or pharmacological inhibition of LSD1 upregulated FOXO3a expression. Specifically, LSD1 interacted with demethylated FOXO3a. The LSD1-demethylated FOXO3a degraded via an ubiquitin-proteasome pathway. Biologically, LSD1 destabilized FOXO3a to abrogate its functions in the suppression of soft agar colony and cell proliferation formation in HO8910 ovarian cancer cells. Knockdown of FOXO3a rescued the restricted cell proliferation by LSD1 downregulation. As a whole, our study clarifies a way in ovarian cancer cell growth through the negative regulation of FOXO3a by LSD1.

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