Molecular Biology Function Of Thyroid Hormone Receptor Associated Protein 3

Background Autophagy is a process that eats up its cytoplasmic protein or organtocytator and makes its bags enter vesicles,and it is used to merge with lysosomes to form autophagy enzymes,which is confirmed by an important way of death of tumor cells,but the role of autophagy in tumor formation and development is still not entirely clear.T3 promotes the growth,development and metabolism of the body,and the combination of the nucleus and the nuclear receptor of the nucleus of the body.The thyroid hormone receptor associated protein 3 is a nuclear receptor coactivation factor,which interacts with multiple nuclear receptors in a ligand dependency,and is stimulated by the inclusion of the thyroid hormone receptor associated protein mediation complex to the hormone response promoter region.However,the regulation mechanism between the T3 and thyroid hormone receptor,THRAP3 is still not fully explained.The study of the related treatment of tumor disease is still further exploration,the recurrence rate of tumor disease is low,the cure rate is low,the disease is serious harm,the current treatment mainly use surgical removal,radiotherapy or chemotherapy,but the recurrence of chemotherapy and postoperative tumor and the risk of the human body are still subject to the debate.The destruction of the cell’s energy metabolism has had a significant impact on cell damage and cancer cells,and the THRAP3 expression in the liver,gastrointestinal tract and breast cancer cells is different,and according to the literature report,the treatment of liver cell tumors can be used in the treatment of liver cells,so we suspect that there is a correlation between the T3,the tumor and the autophagy,and whether the intercorrelation of THRAP3 can be the therapeutic target of the value of the research.And by studying a lot of literature we propose a scientific hypothesis that can T3 affect the cell autophagy by regulating THRAP3.MethodsWe use the Western blotting method to detect the expression of protein,which is a hybrid technique that combines high score discrimination rate gel electrophoresis and immunochemical analysis technology.Using immunofluorescence detection protein positioning;Real-time quantitative PCR is a method of measuring the total amount of the product of the polymerase chain in the DNA amplification reaction.Finally,we completed the research on the subject of gene editing,high throughput sequencing,flow cytometry and enzyme marker detection.ResultsThe ratio size of the cell autophagy can be used to assess the level of autophagy,and found that the ratio of the increase in the level of THRAP3 levels is increased,and then we use the T3 processing cells of different concentration gradients,and we find that with the increase of the T3 concentration gradient and the level of THRAP3 decreases,and then we start to explore the degradation pathway of THRAP3,which is the decrease of synthesis and the increase of degradation.We used a high concentration of T3 processing cells and the joint CHX,MG132 processing cells,and found that cells that were treated with a high concentration of T3 were compared to the cells that were processed after T3 joint MG132,and the levels of the original THRAP3 increased.We further explore the degradation mechanism of THRAP3 and expect to find more.At the same time,we used CRISPR/Cas9 gene editing technology to build the Hep G2 cell THRAP3 gene,which was able to identify genotype and detection protein expression by sequencing,confirming the success of the cell series.When the cell was synchronized to the G2/M period,the detection cell cycle found that the reduction of the amount of cells in the G1 period compared to the WT in the G1 period,using CCK-8 detection cell proliferation found to inhibit the growth of Hep G2 cells in comparison to the WT.Finally,the results of the data difference gene analysis of high-throughput sequencing data were analyzed,and the genes and cancer related genes were selected by the sequence of the transcription group of the cell line,and the correlation gene difference was found by the QPCR test.ConclusionTo sum up,we hope to further explore the potential molecular mechanisms between T3 and autophagism and THRAP3,providing more theoretical basis for the occurrence,development and treatment of related tumor diseases.