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. 2021 May 1;22(5):1523-1529.
doi: 10.31557/APJCP.2021.22.5.1523.

Cytotoxicity Studies of the Crude venom and Fractions of Persian Gulf Snail (Conus textile) on Chronic Lymphocytic Leukemia and Normal Lymphocytes

Ahmad Salimi  1   2 , Shayan Salehian  3 , Akram Aboutorabi  4 , Amir Vazirizadeh  5 , Vahed Adhami  2 , Seyyed Hossein Sajjadi Alehashem  2 , Enayatollah Seydi  6   7 , Jalal Pourahmad  3
Affiliations

Cytotoxicity Studies of the Crude venom and Fractions of Persian Gulf Snail (Conus textile) on Chronic Lymphocytic Leukemia and Normal Lymphocytes

Ahmad Salimi et al. Asian Pac J Cancer Prev. .

Abstract

Background: Marine animals have been considered by many researchers due to their various pharmacological effects. One group of marine animals that have been studied is cone snails. The conotoxin obtained from these marine animals has various therapeutic effects.

Methods: This study was designed to investigate the apoptotic effects of crude venom of Conus textile and its fractions (A and B) on chronic lymphocytic leukemia (CLL) cells. Accordingly, parameters such as cell viability, reactive oxygen species (ROS) level, collapse in mitochondrial membrane potential (MMP), lysosomal membrane damage and caspase-3 activation were evaluated.

Results: The results showed that the crude venom (50, 100 and 200 µg/ml) from Conus textile and its fraction B (50, 100 and 200 µg/ml) significantly reduced viability in CLL B-lymphocyte. In addition, exposure of CLL B-lymphocyte to fraction B (50, 100 and 200 µg/ml) was associated with an increase in the level of ROS, the collapse of the MMP, damage to the lysosomal membrane, and activation of caspase-3.

Conclusion: According to results, it was concluded that fraction B from crude venom of Conus textile causes selective toxicity on CLL B-lymphocyte with almost no effect on a normal lymphocyte. Furthermore, this venom fraction could be a promising candidate for induction of apoptosis in patients with CLL through the mitochondrial pathway.

Keywords: Apoptosis; B-Lymphocyte; Conus textile; mitochondria; oxidative stress.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
Gel Filtration Diagram of Conus Textile Crude Venom (100 mg), Dissolved Venom in Ammonium Acetate Buffer and Loaded on a Sephadex G50 Column, Previously Equilibrated and Eluted with 0.2 M Ammonium Acetate, pH 7.5, and Detected at 280 nm
Figure 2
Figure 2
Cell Viability Assay. (A) Effect of crude venom on CLL B- lymphocytes in 12 hours, (B) effect of fraction A on CLL B- lymphocytes in 12 hours, (C) effect of fraction B on CLL B- lymphocytes in 12 hours, (D) effect of fraction B on normal cells in 12 hours. All data were presented as the mean ± S.D (n=3). *** p<0.001 significantly different from the control group
Figure 3
Figure 3
ROS Assay. Effect of fraction B (50, 100 and 200 µg/ml) on ROS generation in CLL B- lymphocytes. All data were presented as the mean ± S.D (n=3). *** p<0.001 significantly different from the control group. DCF-DA: Dichlorofluorescin Diacetate. FCS: Forward Channel Scatter. FL1: Fluorescence Channel 1
Figure 4.
Figure 4.
MMP Assay. Effect of fraction B (50, 100 and 200 µg/ml) on MMP collapse in CLL B- lymphocytes. All data were presented as the mean ± S.D (n=3). *** p<0.001 significantly different from the control group. FCS, Forward Channel Scatter; FL1, Fluorescence Channel 1
Figure 5
Figure 5
Lysosomal Damage Assay. Effect of fraction B (50, 100 and 200 µg/ml) on lysosomal damage in CLL B- lymphocytes. All data were presented as the mean ± S.D (n=3). *** p<0.001 significantly different from the control group. FCS, Forward Channel Scatter; FL1, Fluorescence Channel 1; AO, Acridine Orange
Figure 6
Figure 6
Caspase-3 Activity Assay. Effect of fraction B (50, 100 and 200 µg/ml) on caspae-3 activity in CLL B- lymphocytes. All data were presented as the mean ± S.D (n=3). * p<0.05 and *** p<0.001 significantly different from the control group
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