Cellular apoptosis: An alternative mechanism of action for caspofungin against Candida glabrata

Date

2019-06

Authors

Aryamloo, Parisa
Asgarian-Omran, Hossein
Aslani, Narges
Hossein-Nataj, Hadi
Shokohi, Tahereh
Badali, Hamid
Nabili, Mojtaba
Abdollahi Gohar, Atefeh
Moazeni, Maryam

Journal Title

Journal ISSN

Volume Title

Publisher

Mazandaran University of Medical Sciences

Abstract

Background and Purpose: Although the mechanism of action for echinocandins is known, the physiological mechanisms by which these antifungal agents cause cell death via the classical apoptotic pathways are not well-defined yet. Regarding this, the present study aimed to evaluate the mechanisms of caspofungin-induced Candida glabrata cell death.

Materials and Methods: For the purpose of the study, the minimum inhibitory concentration (MIC) of caspofungin against C. glabrata (ATCC 90030) was determined using the broth microdilution reference method (CLSI M27-A2 and M27-S4). The annexin V and propidium iodide staining was performed to determine the way through which caspofungin acts against C. glabrata (i.e., through the induction of apoptosis and/or necrosis). Additionally, the possible effect of caspofungin on inducing the expression of two apoptotic genes, namely MCA1 and NUC, was studied using the real-time polymerase chain reaction assay.

Results: According to the obtained MIC value (0.5 μg/mL), C. glabrata, exposed to 0.25, 0.5, and 1 μg/mL of caspofungin, exhibited the features of late apoptosis/necrosis after 18 h of incubation. Furthermore, the use of 0.25, 0.5, and 1 μg/ml caspofungin induced apoptosis (early/late) in 14.67%, 17.04%, and 15.89% of the cells, respectively. The results showed a significant difference between the percentages of early-apoptotic cells at the three concentrations (p <0.05). In addition, the rate of necrosis was significantly greater than that of apoptosis in response to caspofungin. Accordingly, necrosis occurred in 71.26%, 71.26%, and 61.26% of the cells at the caspofungin concentrations of 0.25, 0.5, and 1 μg/mL, respectively (p <0.05). The analysis of the data in the REST software demonstrated a significant increase in the expression of MCA1 and NUC1 genes (p <0.05).

Conclusion: As the findings of the present study indicated, caspofungin promoted both necrosis and apoptosis of C. glabrata cells at concentrations higher than or equal to the MIC value.

Description

Keywords

Candida glabrata, caspofungin, flow cytometry, MCA1, NUC1

Citation

Aryamloo, P., Asgarian-Omran, H., Aslani, N., Hossein-Nataj, H., Shokohi, T., Badali, H., . . . Moazeni, M. (2019). Cellular apoptosis: An alternative mechanism of action for caspofungin against Candida glabrata. Current Medical Mycology, 5(2), 9-15. doi:10.18502/cmm.5.2.1155

Department

Molecular Microbiology and Immunology