Role of Bm30kc6 gene in cell apoptosis and the silk gland degradation signaling pathway in Bombyx mori L

Arch Insect Biochem Physiol. 2020 Nov;105(3):e21741. doi: 10.1002/arch.21741. Epub 2020 Oct 1.

Abstract

Apoptosis is a process of programmed cell death that is regulated by genes independently. The Bm30kc6 gene is a kind of small molecular lipoprotein about 30 kDa, expressed highly in the late stage of the silkworm hemolymph. Our study showed that overexpression of Bm30kc6 could decrease caspase-3 activation. Meanwhile, activation of caspase-3 increased when Bm30kc6 expression was disturbed by small interfering RNA (siRNA). Cell apoptosis was decreased when Bm30kc6 was overexpressed under UV treatment. The apoptosis rate induced by actinomycin D is similar to the trend by UV. It was inferred that Bm30kc6 has an inhibitory effect on the apoptosis of silkworm cells. The apoptosis-related genes, such as BmFadd, BmDredd, and BmDaxx were increased after overexpression of Bm30kc6 or decreased after interference of siRNA. It was speculated that there was an interactive relationship between Bm30kc6, BmDaxx, BmFadd, and BmDredd in the apoptosis signaling pathways. We investigated the transcription expression of the Bm30kc6 gene in different growth stages and tissues of the silkworm. The results showed that Bm30kc6 reached its peak in the hemolymph during the 6th to 7th days of the 5th instar, or in spinning post 24 h of the silk gland. In the silkworm BmN cells treated with caspase-3/7 inhibitor, the caspase-3 enzyme activity, and the expression levels of Bm30kc6, BmFadd, BmDredd, and BmDaxx were significantly reduced. The expression levels of Bm30kc6 increased sharply when silkworms were treated by molting hormone at Day 3 or 5 of the 5th instar. The results indicated that the expression of the Bm30kc6 gene was affected by the molting hormone and was likely to be its downstream target. In conclusion, the results suggest that the Bm30kc6 gene is involved in the regulation of the apoptotic signaling pathway and plays a role in the apoptotic process.

Keywords: Bm30kc6; RNAi; apoptosis; overexpression; silkworm Bombyx mori.

MeSH terms

  • Animals
  • Apoptosis / genetics*
  • Bombyx / genetics*
  • Bombyx / growth & development*
  • Bombyx / metabolism
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Caspase Inhibitors / pharmacology
  • Dactinomycin / pharmacology
  • Ecdysone / pharmacology
  • Gene Expression Regulation, Developmental
  • Hemolymph / metabolism
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • RNA, Small Interfering
  • Signal Transduction
  • Ultraviolet Rays

Substances

  • Caspase Inhibitors
  • Insect Proteins
  • RNA, Small Interfering
  • Dactinomycin
  • Ecdysone
  • Caspase 3