4t-CHQ a Spiro-Quinazolinone Benzenesulfonamide Derivative Induces G0/G1 Cell Cycle arrest and Triggers Apoptosis Through Down-Regulation of Survivin and Bcl2 in the Leukemia Stem-Like KG1-a Cells

Author(s):

Arezoo Rahimian, Majid Mahdavi*, Reza Rahbarghazi and Hojjatollah N. Charoudeh   Pages 1340 - 1349 ( 10 )

Abstract:

Objective: Many experiments have revealed the anti-tumor activity of spiro-quinazolinone derivatives on different cell types. Exposing KG1-a cells to N-(4- tert- butyl- 4&#039;- oxo- 1&#039;H- spiro [cyclohexane- 1, 2&#039;- quinazoline]- 3&#039;(4&#039;H)- yl)- 4- methyl benzenesulfonamide (4t-CHQ), as an active sub-component of spiroquinazolinone benzenesulfonamides, the experiment investigated the possible mechanisms that manifest the role of 4t-CHQ in leukemic KG1-a progenitor cells. Mechanistically, the inhibitory effects of 4t-CHQ on KG1-a cells emerge from its modulating function on the expression of Bax/Bcl2 and survinin proteins. <p></p> Methods: Cell viability was assessed using MTT assay. The IC50 value of cells was calculated to be 131.3μM, after 72h-incubation with 4t-CHQ, ranging from 10 to 150μM. Apoptotic changes were studied using Acridine Orange/Ethidium Bromide (AO/EB) staining. DNA fragmentation was analyzed by agarose gel electrophoresis method. To evaluate the percentage of apoptotic cells and cell growth dynamic apoptotic features, we performed Annexin V/PI double staining assay and cell cycle analysis by flow cytometry. <p></p> Results: According to the results, apoptosis induction was initiated by 4t-CHQ in the KG1-a cells (at IC50 value). Cell dynamic analysis revealed that the cell cycle at the G1 phase was arrested after treatment with 4t- CHQ. Western blotting analysis showed enhancement in the expression ratio of Bax/Bcl-2, while the expression of survinin protein decreased in a time-dependent manner in the KG1-a cells. According to the docking simulation data, the effectiveness of 4t-CHQ on KG1-a cells commenced by its reactions with the functional domain of BH3 and Bcl2 and BIR domains of survivin protein. <p></p> Conclusion: These results demonstrate a remarkable role of 4t- CHQ in arresting leukemia KG1-a stem cells both by induction of apoptosis as well as by down-regulating survivin and Bcl2 proteins.

Keywords:

Apoptosis, spiro-quinazolinone, Bcl2, survivin, KG1-a cell, leukemia.

Affiliation:

Department of Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Department of Biology, Faculty of Natural Sciences, University of Tabriz, Tabriz, Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz

Graphical Abstract:

Read Full-Text article