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Experimental Study on Arsenic Trisulfide Induced Apoptosis and Necrosis of HL-60 Cells

wallpapers News 2021-12-01
Experimental Study on Arsenic Trisulfide Induced Apoptosis and Necrosis of HL-60 Cells
In order to study the role of PML RARα and RARα fusion protein in the apoptosis of human promyelocytic leukemia cell line NB4 induced by arsenic trisulfide (As2S3), this study used cell morphology observation, flow cytometry determination and DNA electrophoresis To observe the apoptosis-inducing effect of arsenic trisulfide on NB4 cells, and use chromosome G banding technology, reverse transcription PCR and Western blotting to determine the changes of PML RARα fusion gene and its expression product, and wild-type RARα protein during this process.The results showed that arsenic trisulfide can induce apoptosis in NB4 cells between 0.5 and 2 μmol/L. During this process, the PML RARα fusion gene did not change significantly, but the PML RARα fusion protein and wild-type RARα protein were significantly reduced.
Bortezomib (bortezomib) and arsenic disulfide (As2S2) inhibit the proliferation of chronic myeloid leukemia (CML) cell line K562 and the homologous cell line K562R resistant to imatinib, and induce apoptosis Its mechanism of action. Methods: Application of tetramethylazolium salt colorimetric method (MTT) proliferation inhibition test, Wright staining cell morphology observation method, Annexin Ⅴ (Annexin Ⅴ) flow cytometry to detect bortezomib and As2S2 Used alone or in combination to inhibit the proliferation and apoptosis of CML cell lines; Western blot to detect the effects of drugs on Bcr-Abl protein and apoptosis-related proteins; reverse transcription PCR (RT-PCR) to detect Ber -Abl mRNA expression. Results: Bortezomib combined with As2S2 can effectively inhibit the proliferation of CML cells, 6 nmol/L bortezomib and 3 μmol/L As2S2 combined with K562 cells for 48 h, the cell growth inhibition rate reached (76.4±3.9 )%, and the proportion of necrotic and apoptotic cells reached 54.0%, which was statistically significant (P<0.05) compared with the single-agent effect (bortezomib 13.9%; As2S2 8.2%). 6 nmol/L bortezomib Used in combination with 3 μmol/L As2S2 for K562R cells for 48 hours, the cell growth inhibition rate was only (55.7±5.5)%. 12 nmol/L bortezomib and 6 μmol/L As2S2 were used in combination with K562R cells for 48 hours, necrosis and apoptosis The proportion of dead cells was 38.0%, which was significantly higher than that of single drugs (bortezomib 24.0%; As2S2 11.2%) (P<0.05). The combined action of drugs synergistically induces apoptosis of CML cells through the mitochondrial pathway, and makes the Bcr of K562 and K562R cells. -Abl protein expression and protein phosphorylation levels decreased.
 

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