# Impact of trehalose and hydroxychloroquine on the function of BRAF (V600E)-siRNA in the A375 melanoma cells

## TL;DR

This study explores how trehalose and hydroxychloroquine affect BRAF (V600E)-siRNA function in melanoma cells, finding that hydroxychloroquine enhances apoptosis and BRAF suppression.

## Contribution

The study introduces a novel combination therapy approach using hydroxychloroquine to improve BRAF (V600E)-siRNA efficacy in melanoma.

## Key findings

- Hydroxychloroquine reduced cell viability and increased apoptosis more than trehalose in A375 cells.
- Combined HCQ and BRAF (V600E)-siRNA significantly down-regulated BRAF and up-regulated CASP3 expression.
- TRE showed limited efficacy and may counteract the benefits of HCQ in promoting apoptosis.

## Abstract

Melanoma, a lethal form of skin cancer, is closely linked to mutations in melanocytes. Due to increased resistance to chemotherapy, innovative strategies, including gene and combination therapies, are being explored. This study evaluates the effects of trehalose (TRE) and hydroxychloroquine (HCQ) in enhancing the efficacy of BRAF (V600E)-siRNA in A375 cells.

The A375 cells were treated, and changes in cell viability were assessed using MTT assays. Apoptosis was evaluated using flow cytometry. Additionally, gene expression analysis of B-Raf proto-oncogene, serine/threonine kinase (BRAF), Caspase 3 (CASP3), and Phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3) was performed using quantitative RT-PCR.

HCQ treatment reduced cell viability and increased apoptosis compared to both control cells and cells treated with TRE. Gene expression analysis showed a significant down-regulation of BRAF expression in cells treated with HCQ and BRAF (V600E)-siRNA compared to siRNA-only treated cells. CASP3 expression was significantly up-regulated in cells treated with combined HCQ and siRNA, indicating a stronger apoptotic response. PIK3R3 expression showed no significant change in the transfected groups.

TRE, either alone or combined with siRNA, showed limited efficacy and may counteract the apoptotic benefits of HCQ. Conversely, HCQ, whether used alone or in combination with siRNA, enhanced apoptosis, suggesting promise as a potential treatment for melanoma.