Effect Of Overexpression Of Bmstat On Silkworm Cell Proliferation And Transcriptome

The JAK/STAT signaling is a multiple cytokines shared pathway. Despite the number and diversity of cytokines, the related proteins and fundamental functions appear to be highly conserved. In order to well study the JAK-STAT pathway in Bombyx mori effecting on cell proliferation, We constructed the transgenic vector pIZT-STAT, and transfected it into BmN cells of Bombyx mori. With continuously screening for three months after the adoption of Zecon, the transformed cells were obtained。Then the cell proliferation was assessed by MTT and flow cytometry. To better understand the JAK-STAT signal pathway, high-throughput transcriptome sequencing technology (RNA-Seq) was used to study comparative transcriptome of the stable transformed cells and normal BmN cells.The experimental results showed that when BmSTAT was over espressed, the cell proliferation rate was speeded, and the expression of level of the development related genes were up-regulated. The results of comparison of the transcriptome between stably transformed cells and normal BmN cells were as below:10853expresed genes were detected in the normal cells. Compared with the expression level of its actin A3, there are41high expression genes (Fold change>2). The productions of these genes are mainly involved in protein synthesis, innate immunity, signal transduction, environmental adaptation.10129expresed genes were detected in stably transformed cells. There were252differently expressed genes, which meted obvious fold change (only expressing in one sample of normal cells or stably transformed cells).123genes were up-regulated,129genes were down-regulated. However,800DEGs were detected, which meted T-test, expressing in both samples of of normal cells and stably transformed cells Among which787genes were up-regulated,13genes were down-regulated. The results of GO analysis showed that the up-regulated genes were mainly involved in cells, cell components, macromolecular complex, organelle, metabolic process, cellular process, catalytic activity, binding et al. The analysis of KEGG signal pathway showed up-regulated genes were mainly involved in carbon metabolism, amino acid synthesis, oxidative phosphorylation, purine metabolism, pyrimidine metabolism, spliceosome, ribosome, RNA transport, protein processing in endoplasmic reticulum, transcription, translation, Cell cycle and apoptosis et al.. The genes related to Ras, Rapl, MAPK, Wnt, Hippo, JAK/STAT, FoXo, Ca2+, P13-AKt, mTOR signaling pathway have been increased. We found that14up regulated genes related to the regulation of cell cycle. The results of GO analysis showed that the down-regulated genes were involved in cell, cell part, metabolic process, obsolete biological process, cellular process, catalytic activity, transporter activity et al.. The analysis of KEGG signal pathway showed down-regulated genes were involved in oxidative phosphorylatio, methane metabolism, drug metabolism-other enzymes, et al. JAK-STAT signaling pathway plays an important role in the regulation of cell growth and proliferation.The normal BmN cells and stable transformation of BmN cells, which overexpression of BmSTAT, were infected with BmCPV polyhedrosis lysate. The expression level of VP7was detected by by Western blotting in days2,6, respectively. Obvious change was not found between the stably transformed cells and normal cells. This result indicates the silkworm JAK/STAT signal pathway had no effection on BmCPV infection.