我们近来发现胸苷酸合成酶（TYMS）在复层细胞中呈稳定的非细胞周期依赖

We have recently discovered that thymidylate synthase (TYMS) exhibits a stable cell cycle-independent nuclear localization in stratified cells. This is obviously different from the previous view that TYMS was SUMO-modified and translocated in the G1 / S phase and located in the S-phase cell nucleus to participate in the de novo synthesis of dTMP. We thus speculate that the difference in the expression of SUMOylation-related molecules in different types of cells may be the molecular mechanism that causes the different nuclear localization of TYMS. To this end, this project intends to first use RT-PCR, immunohistochemistry / immunofluorescence technology to detect the expression differences and subcellular localization of molecules related to SUMO modification in human tissue samples and stratified cells, monolayer cells, and screen directly for TYMS After related SUMO-modified molecules, construct the corresponding molecules and their microRNA eukaryotic expression vectors respectively, and use gene transfection and (or) RNA interference technology to transfer them into stratified and monolayer cells to upregulate or silence SUMO, respectively Modified related molecules, and then used immunofluorescence, immunoprecipitation and mass spectrometry to study their correlation with TYMS subcellular localization, and finally used Balb / c mice to verify, in order to clarify that SUMO chemical modification regulates TYMS nuclear translocation in different cell types Molecular mechanism.

We have recently found that thympathisal synthase (TYMS) presents a stable non-cell cycle-dependent nucleus in complex cells. This is significantly different from the previous view that TYMS was modified by SUMO to be nuclearally transpositioned in the G1/S phase and positioned in the s-stage cell nucleus to participate in dTMP synthesis from the head. We conclude by suggesting that differences in expression of THE SUMO-modified related molecules in different types of cells may be the molecular mechanism that causes TYMS to differ in approved bits. To this end, this project aims to first use RT-PCR, immunohistization/ immunofluorescent technology to detect human tissue samples and complex cells, single-layer cells in SUMO-modifying related molecules expression differences and subcellular positioning, screening out the DIRECTly related to TYMS SUMO modified molecules, respectively, to build the corresponding molecules and microRNA eukaryotic expression vector, and the use of gene transfection And/or RNA interference technology transferred it to complex and single-layer cells to raise or silence SUMO-modifying related molecules, and then use immunofluorescent, immunoprecipitation and mass spectrometry technology to study its correlation with TYMS subcellular positioning, and finally use Balb/c mice to verify, with a view to clarifying the SUMO modification to regulate the molecular mechanism of TYMS nuclear translocation in different types of cells.

Recently, we found that TYMS has a stable non cell cycle dependent nuclear localization in multilayered cells. This is different from the previous view that TYMS was modified by SUMO in G1 / S phase and located in S phase to participate in the de novo synthesis of dTMP. We speculated that the different expression of SUMO related molecules in different types of cells might be the molecular mechanism of different nuclear localization of TYMS. In this project, RT-PCR, immunohistochemistry / immunofluorescence technology are used to detect the expression difference and subcellular localization of SUMO modified molecules in human tissue samples, multilayered cells and monolayer cells. After selecting the sumo modified molecules directly related to TYMS, corresponding eukaryotic expression vectors of molecules and microRNA are constructed respectively, and gene transfection and / or RNA stem are used In order to elucidate the molecular mechanism of SUMO modification in the regulation of TYMS nuclear translocation in different cell types, BALB / c mice were used to test the correlation between SUMO modification and TYMS subcellular localization.