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[Effects of and Knockout on Ferroptosis Sensitivity of RPMI-8226 Cells]. | LitMetric

[Effects of and Knockout on Ferroptosis Sensitivity of RPMI-8226 Cells].

Zhongguo Shi Yan Xue Ye Xue Za Zhi

Institute of Hematology, Xuzhou Medical University; Department of Hematology, Xuzhou Medical University Affiliated Hospital, Xuzhou 221002, Jiangsu Province, China.

Published: October 2024

AI Article Synopsis

  • - The study aimed to explore how the
  • genes affect the sensitivity of multiple myeloma (MM) cells to a process called ferroptosis, using the RPMI-8226 cell line as a model.
  • - Researchers utilized CRISPR/Cas9 technology to knock out these key
  • genes and examined subsequent changes in cellular characteristics, including autophagy-related protein levels and cell viability under different drug conditions.
  • - Results indicated that knocking out the
  • genes increased cell viability in response to ferroptosis-inducing agents, suggesting that the LAP pathway may play a role in regulating this process in MM cells.

Article Abstract

Objective: To investigate the effects of and genes on the sensitivity of multiple myeloma (MM) cell line RPMI-8226 cells to ferroptosis.

Methods: CRISPR/Cas9 technology was used to knock out the autophagy key genes and in RPMI-8226 cells. Western blot was used to identify gene knockout cells, and detect the expression changes of autophagy-related proteins P62 and LC3B. Flow cytometry was used to detect the change of sensitivity of gene knockout cells to RSL3. The content of intracellular ferrous ions and reactive oxygen species (ROS) level in gene knockout cells were detected.

Results: Western blot result confirmed that and genes were knocked out successfully in RPMI-8226 cells. The result of flow cytometry showed that the cell viability of RPMI-8226 cells was dose-dependent on different concentrations of RSL3 ( =-0.969). RSL3 (10 μmol/L) was used to induce ferroptosis in cells of control group and gene knockout groups, then the cell viability in gene knockout groups were both higher than control group after 48 hours (both < 0.001). After knocking out the and genes, the content of intracellular Fe decreased significantly compared with control group (both < 0.01), and the ROS level also decreased (both < 0.001).

Conclusion: Knockout of and genes can inhibit the ferroptosis of MM cells, and LAP pathway may be involved in the regulation.

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Source
http://dx.doi.org/10.19746/j.cnki.issn.1009-2137.2024.05.022DOI Listing

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