Arwa Alkaraki1,2, Walhan Alshaer3, Suha Wehaibi1, Lobna Gharaibeh4, Duaa Abuarqoub1,5, Dana A Alqudah1, Hafsa Al-Azzawi1, Hadil Zureigat6, Mamoun Souleiman6, Abdalla Awidi7,8,9. 1. Cell Therapy Center, The University of Jordan, PO Box: 5825, Amman, 11942, Jordan. 2. Department of Pharmacology, Faculty of Medicine, The University of Jordan, Amman, 11942, Jordan. 3. Cell Therapy Center, The University of Jordan, PO Box: 5825, Amman, 11942, Jordan. walhan.alshaer@ju.edu.jo. 4. Department of Pharmaceutical Sciences, Faculty of Pharmacy, The University of Jordan, Amman, Jordan. 5. Department of Biomedical Sciences and Pharmacology, Faculty of Pharmacy, University of Petra, Amman, 11180, Jordan. 6. Faculty of Medicine, The University of Jordan, Amman, 11942, Jordan. 7. Cell Therapy Center, The University of Jordan, PO Box: 5825, Amman, 11942, Jordan. abdalla.awidi@gmail.com. 8. Faculty of Medicine, The University of Jordan, Amman, 11942, Jordan. abdalla.awidi@gmail.com. 9. Department of Hematology and Oncology, Jordan University Hospital, The University of Jordan, Amman, 11942, Jordan. abdalla.awidi@gmail.com.
Abstract
BACKGROUND/ OBJECTIVE: The absence of receptors in triple-negative breast cancer limits therapeutic choices utilized in clinical management of the disease. Doxorubicin is an important member of therapeutic regimens that is hindered by emergence of resistance. The current work aim to investigate of therapeutic potential of single and combinations of siRNA molecules designed for silencing STAT 3, Notch-1, and β-catenin genes in wild type and doxorubicin resistant MDA-MB-231 triple negative breast cancer cell line. METHODS: Doxorubicin resistant MDA-MB-231 cell line was developed and characterized for the expression of multidrug resistance-related genes, CD44/CD24 markers, inflammatory cytokines, and the expression of STAT 3, Notch-1, and β-catenin targeted genes. Further, the effect of single and combinations of siRNA on cell viability and chemosensitivity of both wild type MDA-MB-231 cells (MDA-MB-231/WT) and doxorubicin resistant MDA-MB-231 cells (MDA-MB-231/DR250) were assessed by MTT assay. RESULTS: The IC50 of doxorubicin was 10-folds higher in MDA-MB-231/DR250 resistant cells compared to MDA-MB-231/WT control cells, 1.53 ± 0.24 μM compared to 0.16 ± 0.02 μM, respectively. The expression of targeted genes was higher in resistant cells compared to control cells, 3.6 ± 0.16 folds increase in β-catenin, 2.7 ± 0.09 folds increase in Notch-1, and 1.8 ± 0.09 folds increase in STAT-3. Following treatment with siRNAs, there was a variable reduction in mRNA expression of each of the targeted genes compared to scrambled siRNA and a reduction in IC50 in both cell lines. The effect of a combination of three genes produced the largest reduction in IC50 in resistant cell line. CONCLUSION: Our study showed that the silencing of single and multiple genes involved in drug resistance and tumor progression by siRNA can enhance the chemosensitivity of cancer cells to conventional chemotherapy.
BACKGROUND/ OBJECTIVE: The absence of receptors in triple-negative breast cancer limits therapeutic choices utilized in clinical management of the disease. Doxorubicin is an important member of therapeutic regimens that is hindered by emergence of resistance. The current work aim to investigate of therapeutic potential of single and combinations of siRNA molecules designed for silencing STAT 3, Notch-1, and β-catenin genes in wild type and doxorubicin resistant MDA-MB-231 triple negative breast cancer cell line. METHODS:Doxorubicin resistant MDA-MB-231 cell line was developed and characterized for the expression of multidrug resistance-related genes, CD44/CD24 markers, inflammatory cytokines, and the expression of STAT 3, Notch-1, and β-catenin targeted genes. Further, the effect of single and combinations of siRNA on cell viability and chemosensitivity of both wild type MDA-MB-231 cells (MDA-MB-231/WT) and doxorubicin resistant MDA-MB-231 cells (MDA-MB-231/DR250) were assessed by MTT assay. RESULTS: The IC50 of doxorubicin was 10-folds higher in MDA-MB-231/DR250 resistant cells compared to MDA-MB-231/WT control cells, 1.53 ± 0.24 μM compared to 0.16 ± 0.02 μM, respectively. The expression of targeted genes was higher in resistant cells compared to control cells, 3.6 ± 0.16 folds increase in β-catenin, 2.7 ± 0.09 folds increase in Notch-1, and 1.8 ± 0.09 folds increase in STAT-3. Following treatment with siRNAs, there was a variable reduction in mRNA expression of each of the targeted genes compared to scrambled siRNA and a reduction in IC50 in both cell lines. The effect of a combination of three genes produced the largest reduction in IC50 in resistant cell line. CONCLUSION: Our study showed that the silencing of single and multiple genes involved in drug resistance and tumor progression by siRNA can enhance the chemosensitivity of cancer cells to conventional chemotherapy.
Entities:
Keywords:
Drug resistance; Notch 1; STAT 3; Triple negative breast cancer (TNBC); β-Catenin
Authors: Alaa A A Aljabali; Mohammad A Obeid; Hamid A Bakshi; Walhan Alshaer; Raed M Ennab; Bahaa Al-Trad; Wesam Al Khateeb; Khalid M Al-Batayneh; Abdulfattah Al-Kadash; Shrouq Alsotari; Hamdi Nsairat; Murtaza M Tambuwala Journal: Molecules Date: 2022-03-11 Impact factor: 4.411