Chicken Egg White Extract Induces High Expression of tRFs&tiRNAs Molecules in 293T Cells and Its Cell Function Verification
RUAN Guangping, YAO Xiang, HE Jie, MO Ping, WANG Kai, YANG Zailing, PANG Rongqing, ZHU Xiangqing, PAN Xinghua*
tRFs (tRNA-derived fragments) and tiRNAs (tRNA-derived stress-induced RNAs) are derived fragments of tRNAs, which belong to the short non-coding RNA family and participate in complex biological reactions by transcription, translation and signaling pathways. To verify the cell function of the three tRFs&tiRNAs molecules raised after the chicken egg white extract induces 293T cells. 293T cells were added to a 6-well plate, among which 3 wells added with ordinary medium, and 3 wells added with 50% chicken egg white extract medium, and cultured for 3 days. Three samples in the control group and three samples in the induction group were subjected to high-throughput sequencing to detect the differential expression of tRFs&tiRNAs molecules in the two groups. It was verified by testing that the induced cells had 3 tRFs&tiRNAs molecules steadily increasing. The up-regulated expression of these 3 molecules is statistically significant. These three molecules were synthesized and transfected into 293T cells. The changes of pluripotency factors OCT4 and NANOG were detected by WB. The changes of pluripotency genes OCT4 and NANOG and the relative expression of telomeres were detected by quantitative PCR, and the expression variety of pluripotency factors OCT4 and NANOG were detected by flow cytometry. Simultaneously detect the changes of cell proliferation, apoptosis and cell cycle after transfection of these three molecules into 293T cells. The results indicated after the 3 molecules were transfected into 293T cells; the expression of pluripotent factors OCT4 and NANOG was significantly higher than that of untransfected cells detected by WB, and the relative expression levels of pluripotent genes OCT4 and NANOG were significantly higher than that of untransfected cells detected by quantitative PCR. Telomeres are significantly increased compared with untransfected cells. Flow cytometry detected a significant increase in cells expressing pluripotency factors OCT4 and NANOG compared with untransfected cells. After these 3 molecules were transfected into 293T cells, cell viability increased, cell apoptosis decreased, and cell cycle changed to a certain extent. It was proved that the overexpression of these 3 molecules could promote the increase of the expression of pluripotent factors OCT4 and NANOG in 293T cells, promote the growth of telomeres, and make cells younger. At the same time, overexpression of these 3 molecules can increase cell viability and decrease cell apoptosis.
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