Chloride intracellular channel gene knockdown induces insect cell lines death and level increases of intracellular calcium ions

Jiqiang Song; Yanping Yu; Zhichao Yan; Shan Xiao; Xianxin Zhao; Fang Wang; Qi Fang; Gongyin Ye

doi:10.3389/fphys.2023.1217954

Chloride intracellular channel gene knockdown induces insect cell lines death and level increases of intracellular calcium ions

Front Physiol. 2023 Jul 6:14:1217954. doi: 10.3389/fphys.2023.1217954. eCollection 2023.

Authors

Jiqiang Song¹, Yanping Yu¹, Zhichao Yan², Shan Xiao¹, Xianxin Zhao¹, Fang Wang¹, Qi Fang¹, Gongyin Ye¹

Affiliations

¹ State Key Laboratory of Rice Biology and Breeding & Ministry of Agricultural and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, Zhejiang University, Hangzhou, China.
² Department of Entomology, Nanjing Agricultural University, Nanjing, China.

Abstract

Chloride intracellular channel (CLIC) is a member of the chloride channel protein family for which growing evidence supports a pivotal role in fundamental cellular events. However, the physiological function of CLIC in insects is still rarely uncovered. The ovary-derived High Five (Hi-5) cell line isolated from the cabbage looper (Trichoplusia ni) is widely used in laboratories. Here, we studied both characteristics and functions of CLIC in Hi-5 cells (TnCLIC). We identified the TnCLIC gene in Hi-5 cells and annotated highly conserved CLIC proteins in most insect species. After RNA interference of TnCLIC, the phenomenon of significantly increased cell death suggests that the TnCLIC protein is essential for the survival of Hi-5 cells. The same lethal effect was also observed in Spodoptera frugiperda 9 and Drosophila melanogaster Schneider 2 cells after CLIC knockdown. Furthermore, we found that this kind of cell death was accompanied by increases in intracellular calcium ions after TnCLIC knockdown with the transcriptomic analyses and the detection of calcium levels. Our results provide insights into insect CLIC as a key factor for cell survival and lay the foundation for the cell death mechanism.

Keywords: RNA interference; cell death; chloride intracellular channel; insect cell lines; intracellular calcium ions.

Grants and funding

This work was supported by Key Program of National Natural Science Foundation of China (NSFC) (Grant no. 31830074 to GY), the Joint Funds of the National Natural Science Foundation of China (NSFC) (Grant no. U21A20225 to GY), the Program of National Natural Science Foundation of China (Grant no. 32072480 to QF), Zhejiang Provincial Natural Science Foundation of China (Grant no. LTGN23C140001 to QF) and the Fundamental Research Funds for the Central Universities (Grant no. 2021FZZX001-31 to GY and QF).