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. 2023 Jul 1;17(3):177-185.
doi: 10.18502/ijhoscr.v17i3.13307.

The Effect of the Persian Gulf Jellyfish (Cassiopea andromeda) Venom on the Expression of P15, P21, P53, DNMT1, and Bcl-2 in Acute Lymphoblastic Leukemia Jurkat Cells

Reza Dehghani  1   2 , Ali Amrooni  1   2 , Fatemeh Hosseinpour-Soleimani  1   2 , Gholamhossein Mohebbi  3 , Narges Obeidi  2   3
Affiliations

The Effect of the Persian Gulf Jellyfish (Cassiopea andromeda) Venom on the Expression of P15, P21, P53, DNMT1, and Bcl-2 in Acute Lymphoblastic Leukemia Jurkat Cells

Reza Dehghani et al. Int J Hematol Oncol Stem Cell Res. .

Abstract

Background: One of the acute hematologic malignancies is acute lymphoblastic leukemia (ALL), which is formed in B or T lymphocyte stem cells. Regarding the increasing tendency to herbal and marine studies and unclear characteristics of Cassiopea andromeda Venom, this study was performed to determine its effects on Jurkat cells as a model for T-ALL. Materials and Methods: In this experimental study, the cells were treated with a variety of concentrations of Cassiopea andromeda venom at different periods and times. Growth inhibition and toxic effects of Cassiopea andromeda Venom were evaluated by methyl thiazole tetrazolium salt reduction (MTT test). The flow cytometry analysis was carried out using 7-aminoactinomycin D (7AAD) and Annexin V stains to evaluate the venom's effect on apoptotic pathways. Besides, Real-Time PCR was performed to evaluate the relative gene expression. Results: Cassiopea andromeda venom inhibited the growth of Jurkat cells in a concentration and time manner. Jurkat cell growth was inhibited by 48.9% after 72 hours of treatment with 250µg/mL Cassiopea andromeda venom. The venom increased the apoptotic process through the upregulation of p15INK4b and P53 proteins and downregulation of Bcl-2, p21 WAF1/CIP1, and DNMT1 in the Jurkat cell line. Conclusion: Considering the growth inhibitory property of Cassiopea andromeda Venom, we recommend it as a part of combinational medication for treating ALL in animal trials and for other leukemias in vitro studies.

Keywords: Acute lymphoblastic leukemia; Apoptosis; Cnidaria.

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Figures

Figure 1
Figure 1
A) Cell survival comparison in Jurkat and normal lymphocytes at different concentrations in 24h. The difference in survival rate between the different concentrations of Jurkat (P = 0.066) and normal lymphocytes (P = 0.071) were not significant. B) Comparison of Jurkat cell survival and normal lymphocytes at different concentrations in 48h. The difference in survival rate between different concentrations of cell Jurkat (P = 0.999) was not significant and was significant in normal lymphocytes (P = 0.004). C) Comparison of Jurkat cell survival and normal lymphocytes at different concentrations in 72h. The difference in survival rate between the different concentrations of Jurkat (P < 0.001) and normal lymphocytes (P = 0.049) were significant.
Figure 2
Figure 2
Comparison of the effect of IC50 concentration with Cassiopea andromeda Venom in Jurkat cells. (×40) A: Jurkat control cell, B: Jurkat cell after 72 hours of treatment with 250 µg/mL concentration
Figure 3
Figure 3
The apoptotic effect of Cassiopea andromeda Venom (250 µg/mL) on Jurkat cell versus control group after 72 hours. AnnexinV(neg)/7AAD(neg), AnnexinV(pos)/7AAD(neg), AnnexinV(neg)/7AAD(pos), and AnnexinV(pos)/7AAD(pos), were considered as viable, early apoptotic, necrotic and late apoptotic, respectively. A) Control Jurkat. B) Treated Jurkat.
Figure 4
Figure 4
Gene expression fold change in Jurkat cells
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