Screening And Functional Identification Of Long Noncoding RNA Associated With Autophagy In Silkworm

With the deep study of long non-coding RNAs(LncRNAs),the functional identification of LncRNA of silkworm will provide basis for genetic,evolutionary and genomic research of silkworm.The abundance and specificity of LncRNAs have important effects on neurodevelopment,cognitive activity and neurodegenerative processes.Because the molecular mechanism of cell autophagy is conservative in different species,autophagy pathway participates in the occurrence and development of many diseases,and LncRNAs has stage expression patterns.Studying the regulation mechanism of LncRNAs in autophagy signaling pathway of silkworm,which will be of great value and referential significance for the functional research of LncRNAs as a drug target in the diagnosis and treatment of diseases.In this study,the silkworm was the research object.The autophagic changes in silkworm tissues and cell were detected after injecting 20-hydroxyecdysone,also known as molting hormone,into silkworm individuals.Then the LncRNAs related to autophagy were selected by high-throughput sequencing and Real-time Quantitative PCR(RT-qPCR),and the sequence and preliminary function were studied.This study will provide basis for the molecular mechanism of LncRNAs in silkworm autophagy signaling pathway,and provide reference for the research of LncRNAs in diseases and tumors.The results are as follows:(1)Molting hormone treatment and fat body staining showed that molting hormone promoted autophagy in the fat body,and fat body tissue before and after treatmen for different time were used high throughput sequencing of LncRNAs.(2)After real-time quantitative PCR analysis,Lnc560 gene was selected from 10 LncRNAs related to autophagy differently expressed,after molting hormone treatment,the results showed that Lnc560 may be involved in the autophagy process of fat body.(3)Atg4B was predivted as the target gene of Lnc560 by bioinformatics analysis was,and the expression analysis of the Lnc560 gene and predicted target gene Atg4 B showed that they co-expressed in all the samples.(4)The expression of Lnc560 gene in BmN cells treated with molting hormone was detected by real-time quantitative PCR,and the results were consistent with the results of individual experiments.(5)In situ hybridization of Lnc560 gene in BmN cells showed that Lnc560 gene was expressed in both nucleus and cytoplasm.