Knockdown of EIF3G inhibits the intracellular protein translation of H1299 cells
eIF3 is the largest translation initiation factor in mammalian cells, consisting of 13 subunits. This translation initiation factor is involved in multiple processes of protein translation in cells, including translation initiation, termination, and ribosome recycling. Several studies have reported that multiple subunits of eIF3 exhibit abnormal expression in tumor cells and play an important role in the occurrence and development of tumors. In this study, it was found that changes in the expression level of EIF3G significantly affected the growth of non-small cell lung cancer. Knockdown of EIF3G inhibited the intracellular protein translation process of non-small cell lung cancer cells H1299. Through the study of translatome, it was found that knockdown of EIF3G significantly affected the cell cycle processes in H1299 cells. In vitro cell experiments also showed that changes in the expression level of EIF3G influences the cell cycle distribution of H1299 cells. This study is the first to explore the impact of knockdown of EIF3G on the translatome of H1299 cells. Overall design: The H1299 cells treated with siEIF3Gs or siCtrl were lysed, and the cell supernatants were collected after centrifugation. Ten percent of the total lysate was taken as input control. The cell lysate supernatants were then subjected to sucrose density gradient centrifugation, and the resulting products were separated using Gradient108 and Fractionator (BIOCOMP, Canada). The centrifuged products of the polysomal fraction were collected for RNA extraction. Subsequently, both the input control and the mRNA bound to the polysomes were extracted and sequenced.