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Hemp Extract with Specific Anti‐Cancer Properties against Ovarian Cancer
Author(s) -
Sumanasekera Wasana,
Duff Adam,
Biela Sara,
Wang Annie,
Riggs Kayla,
Croley Ashley,
Cox Arthur
Publication year - 2021
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2021.35.s1.02877
Subject(s) - ovarian cancer , cancer , cancer cell , medicine , viability assay , metastasis , cytotoxicity , cancer research , oncology , pharmacology , traditional medicine , cell , biology , in vitro , biochemistry
Ovarian cancer is one of the most aggressive gynecological cancers (Matz et al., 2017) and the fifth leading cause of cancer‐related deaths among women (Siegel et al., 2015; American Cancer Society, 2019). The current treatment options include Cisplatin and Liposomal Doxorubicin. However, due to limited efficacy and unwanted side effects of the existing therapeutics, it is important to look for alternative therapies. Due to growing interest in medicinal treatments with natural substances, we began a search for natural products with anti‐cancer activity. Most of the existing literature is about the use of medical cannabis or Dronabinol for cancer‐related side effects management. Some other studies are beginning to emerge on the anti‐cancer effects of certain cannabinoids or mixtures of cannabinoids on few cancers. However, no one has reported the anti‐cancer properties on Hemp Extract (HE) that we utilized. The objective of this study is to investigate whether HE has anti‐cancer properties against ovarian cancer cells (OCCs) while being safe to normal ovary epithelial cells (NHOECs). Our specific aims were to investigate 1) Dose response relationship and half maximal effective concentration (EC 50) for HE; 2) HE's ability to specifically decrease ovarian cancer cell (OCC) viability; 3) Specifically cause cytotoxicity to OCCs; 4) Specifically decrease OCC death; and 5) Decrease metastasis of OCCs. Our methods include preparation and purification of HE, cannabinoid amount and molarity calculations, cell culture, HE Treatments, EC 50 calculations, and multiple cell‐based assays / detection methods including cell viability assays (MTT, CALCEIN), cell death detection (ETHD‐1), apoptosis assays (CASPASE), phase contrast microscopy, cytotoxicity assays (LDH), and the cell migration assay. The HE was produced using supercritical carbon dioxide extraction method and given to the laboratory. This extract was made from the flowers, stalks, and leaves of the seedless hemp plants. HE stocks and different working concentrations were made and purified in the laboratory. We have treated cultured OCCs (A2780 and MES‐OV) and NHOECs with different concentrations of HE for 24 hours. Cannabinoid amounts and molarity present in each hemp treatment concentration were calculated. Dose repose relationships were established, and EC 50 was calculated for A2780 OCCs treated with HE. Several cell‐based assays were conducted to fulfill each of the specific aims. We found that the EC 50 value of the HE equals to 13.59 µg/ml HE (containing 12.5 µM total CBD), HE caused OCC specific death, caused OCC specific cytotoxicity, and attenuated OCC proliferation while being safe to NHOECs. HE also decreased OCC migration. Here we concluded that HE has specific anti‐cancer activity against ovarian cancer while being safe to NHOECs. To the best of our knowledge, the ovarian cancer‐cell specific anti‐cancer properties of HE has not been reported to date. The mechanisms behind this specific anti‐cancer properties are currently under investigation.