ECE2020 Audio ePoster Presentations Endocrine-related Cancer (14 abstracts)
Reversal effect of Vitamin D (VitD) on exosome-mediated everolimus resistance (EveR) in hepatocellular carcinoma (HCC) cell line: A molecular study
Annalisa Gentile
,
Mariarosaria Negri
,
Tatiana Montò
,
Roberta Patalano
,
Cristina de Angelis
,
Chiara Simeoli
,
Rosario Ferrigno
,
Maria Cristina De Martino
,
Renata Simona Auriemma
,
Annamaria Colao
,
Rosario Pivonello & Claudia Pivonello
Federico II University
,
Clinical Medicine and Surgery
,
Naples
&
Italy
Cancer exosomes emerged as important mediators of cancer drug resistance. Drug exposure-mediated EveR in HCC cell lines can be reversed by VitD pre- and co-treatment. The current study aimed to investigate whether cancer exosomes from EveR HCC cell line are able to induce EveR in parental cells and whether VitD may activate reversal effect on exosome-mediated EveR.
To this purpose, parental JHH-6 and EveR JHH-6 cell line were used. The uptakeof exosomes, staining with membrane dye PKH67, has been evaluated using immunofluorescence (IF). Recipient parental JHH-6 cells were treated for 16 days with exosomes from EveR cells (Exo EveR parental JHH-6) and cell proliferation was evaluated by DNA assay in parental, EveR and Exo EveR parental JHH-6 cells after treatment with everolimus (EVE) at escalating doses (from 10–11 M to 10–7 M) for 6 days, with and without VitD 10–7 M at lowest EVE concentrations used to better visualized the drug effect. Intracellular c-MYC and YAP-1 protein regulation was investigated by western blot (WB) after 6 days of VitD treatment in parental JHH-6, EveR JHH-6 and Exo EveR Parental JHH-6. JHH-6 parental cells were able to uptake exosomes from EveR JHH-6 cells within 18 hrs as shown by IF. Exosome’s uptake conferred EveR to parental JHH-6 cells. This effect was visible in particular at lowest EVE concentrations. Treatment with EVE 10–10 M induced 19.76% of inhibition (P = 0.05 vs control) in parental JHH-6 but no significant inhibition in EveR JHH-6 and Exo EveR parental JHH-6; EVE 10–9 M induced 56.53% (P = 0.0001 vs control) in parental JHH-6, 26.11% of inhibition (P = 0.05 vs control; p = 0.05 vs EVE 10–9 M in parental JHH-6) in Exo EveR parental JHH-6 but not in EveR JHH-6. VitD alone induced 50.19% of inhibition (P = 0.0001 vs control) in parental JHH-6 but no significant inhibition in EveR JHH-6 and Exo EveR parental JHH-6, while co-treatment of VitD with EVE 10–11 M and 10–10 M induced 54.78%, 56.09% and 29.61% of inhibition (P = 0.0001 and P = 0.01 vs EVE 10–11 M alone) and 28.06%, 25.37% and 38.60% of inhibition (P = 0.01, P = 0.05 and P = 0.0001 vs EVE 10–10 M alone)in parental, EveR and Exo EveR parental JHH-6, respectively. Moreover, EveR and Exo EveR parental JHH-6 showed c-Myc and YAP-1 protein upregulation compared to parental JHH-6 but 6 days VitD treatment downregulated their expression. In conclusion, these preliminary data demonstrated that VitD may regulate exosome-mediated drug-resistance mechanisms in JHH-6 cell line, although these mechanisms need to be confirmed in other HCC cell lines.
Article tools
My recent searches
My recently viewed abstracts
Authors
Endocrine Abstracts
ISSN 1470-3947 (print) | ISSN 1479-6848 (online)
© Bioscientifica 2024 |
Privacy policy |
Cookie settings
BiosciAbstracts
Bioscientifica Abstracts is the gateway to a series of products that provide a permanent, citable record of abstracts for biomedical and life science conferences.
Find out more