Inhibition of silkworm vacuolar‐type ATPase activity by its inhibitor Bafilomycin A1 induces caspase‐dependent apoptosis in an embryonic cell line of silkworm
Archives of Insect Biochemistry and Physiology
Published online on September 24, 2018
Abstract
---
- "\nAbstract\nVacuolar‐type ATPase (V‐ATPase) is a type of hydrogen ion transporter
located in the vesicular membrane‐like system, which mediates active transport and
intracellular acidification in various compartments. In mammals, V‐ATPase has been
reported to play a key role in cell proliferation and apoptosis. The studies of
V‐ATPase in silkworm mainly focus on the acidification regulation of midgut and
silk gland and immune resistance. However, there are few reports about the function
of silkworm V‐ATPase on cell proliferation, autophagy, and apoptosis. Thus, the
function of V‐ATPase in a cell line of Bombyx mori (BmE) was investigated by treating
the cell line with bafilomycin A1, a specific inhibitor of V‐ATPase. Cell counting
kit 8 (CCK8) and flow cytometry analysis showed that bafilomycin A1 treatment decreased
the cell proliferation activity, affected the cell cycle progression and induced
cell apoptosis. LysoTracker Red staining showed that the target of bafilomycin A1
is lysosome. The expression of all autophagy‐related genes (\nBmATG5, \nBmATG6,
and \nBmATG8) decreased, indicating that cell autophagy was inhibited. The analysis
of the apoptosis pathway demonstrated that inhibiting the activity of V‐ATPase of
BmE cells could promote mitochondria to release cytochrome C, inhibit the expression
of \nBmIAP, and activate the caspase cascade to induce apoptosis. All these findings
systematically illustrate the effects of V‐ATPase on the proliferation, autophagy,
and apoptosis in BmE cells, and provide new ideas and a theoretical basis for further
study on the function of V‐ATPase in BmE."
- Archives of Insect Biochemistry and Physiology, EarlyView.