CRISPR/Cas9-Mediated Knockout of ZFP36L1 Impairs Cell Proliferation, Alters Cell-Cycle Progression, and Enhances DNA Damage Responses in MDA-MB-231 Triple-Negative Breast Cancer Cells

Zinc finger protein 36-like 1 (ZFP36L1) is a protein that plays a role in regulating gene expression and has been linked to tumor progression and DNA damage responses. However, the impact of ZFP36L1 knockout on triple-negative breast cancer (TNBC) cells is not well understood. This study aimed to examine the consequences of CRISPR/Cas9-mediated ZFP36L1 knockout on cell proliferation, sensitivity to the chemotherapy drug doxorubicin (DOX), cell-cycle progression, and DNA damage responses in MDA-MB-231 TNBC cells.

The researchers created both wild-type (WT) and CRISPR/Cas9-generated ZFP36L1 knockout (KO) MDA-MB-231 cells, which were then cultured under standard conditions over a three-week period. Cell proliferation was measured by counting the number of cells at regular intervals. To assess the effect of DOX treatment on cell viability, the MTT assay was employed, and the half-maximal inhibitory concentration (IC50) values were calculated.

Cell-cycle distribution was determined through propidium iodide flow cytometry performed 24 hours after DOX exposure. DNA damage was quantified using {gamma}-H2AX flow cytometric analysis.

The study findings indicate that the CRISPR/Cas9-mediated ZFP36L1 knockout impaired cell proliferation, altered cell-cycle progression, and enhanced DNA damage responses in MDA-MB-231 TNBC cells.