CD44 Contributes to the Regulation of MDR1 Protein and Doxorubicin Chemoresistance in Osteosarcoma.
Monserrat Gerardo-RamírezFriederike L KeggenhoffVanessa GiamDiana BeckerMarco GrothNils HartmannBeate Katharina StraubHelen MorrisonPeter Robert GalleJens-Uwe MarquardtPeter HerrlichMonika HartmannPublished in: International journal of molecular sciences (2022)
Osteosarcoma is the most common type of pediatric bone tumor. Despite great advances in chemotherapy during the past decades, the survival rates of osteosarcoma patients remain unsatisfactory. Drug resistance is one of the main reasons, leading to treatment failure and poor prognosis. Previous reports correlated expression of cluster of differentiation 44 (CD44) with drug resistance and poor survival of osteosarcoma patients, however the underlying mechanisms are poorly defined. Here, we investigated the role of CD44 in the regulation of drug chemoresistance, using osteosarcoma cells isolated from mice carrying a mutation of the tumor suppressor neurofibromatosis type 2 ( Nf2 ) gene. CD44 expression was knocked-down in the cells using CRISPR/Cas9 approach. Subsequently, CD44 isoforms and mutants were re-introduced to investigate CD44-dependent processes. Sensitivity to doxorubicin was analyzed in the osteosarcoma cells with modified CD44 expression by immunoblot, colony formation- and WST-1 assay. To dissect the molecular alterations induced by deletion of Cd44 , RNA sequencing was performed on Cd44 -positive and Cd44 -negative primary osteosarcoma tissues isolated from Nf2 -mutant mice. Subsequently, expression of candidate genes was evaluated by quantitative reverse transcription PCR (qRT-PCR). Our results indicate that CD44 increases the resistance of osteosarcoma cells to doxorubicin by up-regulating the levels of multidrug resistance (MDR) 1 protein expression, and suggest the role of proteolytically released CD44 intracellular domain, and hyaluronan interactions in this process. Moreover, high throughput sequencing analysis identified differential regulation of several apoptosis-related genes in Cd44 -positive and -negative primary osteosarcomas, including p53 apoptosis effector related to PMP-22 ( Perp ). Deletion of Cd44 in osteosarcoma cells led to doxorubicin-dependent p53 activation and a profound increase in Perp mRNA expression. Overall, our results suggest that CD44 might be an important regulator of drug resistance and suggest that targeting CD44 can sensitize osteosarcoma to standard chemotherapy.
Keyphrases
- poor prognosis
- cell cycle arrest
- induced apoptosis
- nk cells
- crispr cas
- oxidative stress
- emergency department
- type diabetes
- squamous cell carcinoma
- gene expression
- radiation therapy
- skeletal muscle
- high resolution
- endoplasmic reticulum stress
- young adults
- multidrug resistant
- inflammatory response
- single molecule
- high throughput
- ejection fraction
- autism spectrum disorder
- immune response
- regulatory t cells
- postmenopausal women
- replacement therapy
- high fat diet induced
- cancer therapy