Targeting Cdk11 in osteosarcoma cells using the CRISPR‐cas9 system

Abstract

Osteosarcoma is the most common type primary malignant tumor of bone. Patients with regional osteosarcoma are routinely treated with surgery and chemotherapy. In addition, many patients with metastatic or recurrent osteosarcoma show poor prognosis with current chemotherapy agents. Therefore, it is important to improve the general condition and the overall survival rate of patients with osteosarcoma by identifying novel therapeutic strategies. Recent studies have revealed that CDK11 is essential in osteosarcoma cell growth and survival by inhibiting CDK11 mRNA expression with RNAi. Here, we apply the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system, a robust and highly efficient novel genome editing tool, to determine the effect of targeting endogenous CDK11 gene at the DNA level in osteosarcoma cell lines. We show that CDK11 can be efficiently silenced by CRISPR-Cas9. Inhibition of CDK11 is associated with decreased cell proliferation and viability, and induces cell death in osteosarcoma cell lines KHOS and U-2OS. Furthermore, the migration and invasion activities are also markedly reduced by CDK11 knockout. These results demonstrate that CRISPR-Cas9 system is a useful tool for the modification of endogenous CDK11 gene expression, and CRISPR-Cas9 targeted CDK11 knockout may be a promising therapeutic regimen for the treatment of osteosarcoma.

Cyclin-Dependent Kinase 11 (CDK11) Is Required for Ovarian Cancer Cell Growth In Vitro and In Vivo, and Its Inhibition Causes Apoptosis and Sensitizes Cells to Paclitaxel

“…CDK11 may also play a role in cancer cell migration and invasion, as the migration and invasion activities were significantly reduced in CRISPR-Cas9 CDK11 knockout cancer cells (42). Knockdown of CDK11 by CRISPR-based RNAi technology also showed growth-inhibiting effects in mammalian tuberous sclerosis complex 2 (TSC2)-deficient cell lines, including in human tumor-derived acute myeloid leukemia (AML) cells (20).…”

Aplicación de la nanobiotecnología con el sistema CRISPR-cas

“…En líneas celulares de osteosarcoma, el silenciamiento estable de Quinasa dependiente de ciclina (Cdk11), con CRISPR/Cas aumenta la muerte celular, disminuye la migración y reduce la invasión por células malignas (51) ; en otros experimentos, se corrigieron el gen Fah para la tirosinemia43 y el gen Dmd para la distrofia a muscular de Duchenne (52) y en células madre intestinales de pacientes con fibrosis quística, se logró la modificación del receptor de membrana CFTR (Cystic Fibrosis transmenbrane conductance regulator). Estas evidencias confirman que CRISPR/Cas es un sistema viable y eficaz, aplicable en modelos de animales adultos, para el desarrollo de terapias génicas (52) .…”

Development and potential applications of CRISPR-Cas9 genome editing technology in sarcoma

“…Several studies have revealed that CDK11 is essential in cancer, including in sarcoma cell growth and survival [21, 23, 33, 71, 86]. Our previous study identified that CDK11 is critical for the growth and proliferation of liposarcoma cells, suggesting that CDK11 may be a promising therapeutic target for treatment of liposarcoma patients [33].…”

“…Our previous study identified that CDK11 is critical for the growth and proliferation of liposarcoma cells, suggesting that CDK11 may be a promising therapeutic target for treatment of liposarcoma patients [33]. Recently, CRISPR-Cas9 was applied to determine the effect of targeting endogenous CDK11 at the DNA level in osteosarcoma cell lines [23]. Furthermore, the migration and invasion activity was markedly reduced by CDK11 knockout, indicating that CDK11 maybe a novel therapeutic target for osteosarcoma [23].…”

“…Recently, CRISPR-Cas9 was applied to determine the effect of targeting endogenous CDK11 at the DNA level in osteosarcoma cell lines [23]. Furthermore, the migration and invasion activity was markedly reduced by CDK11 knockout, indicating that CDK11 maybe a novel therapeutic target for osteosarcoma [23]. …”