沉默DJ-1对DADS抑制MGC803细胞增殖与迁移侵袭的影响
[Abstract]:Aim: to study the effect of silencing DJ-1 on the proliferation, migration and invasion of MGC803 cells inhibited by DADS. It is confirmed that DJ-1 is one of the targets of DADS in inhibiting migration and invasion of human gastric cancer cells. Methods: DJ-1 silencing plasmid was constructed and transfected into MGC803 cells. Stable DJ-1 silencing MGC803 cell line. Dads was established to treat MGC803 cells. After empty vector group and DJ-1 silencing MGC803 cells, MTT, flat clone formation test, scratch test and Transwell invasion test were used. The effects of DADS and DJ-1 silencing on the proliferation, migration and invasion of MGC803 cells and the tumorigenic ability of nude mice were detected by tumorigenic assay in nude mice. The expression of DJ-1 in MGC803 cells before and after DADS treatment. Western Blot was used to detect the changes of PTEN,Akt,p-Akt expression in MGC803 cells induced by DADS and DJ-1 silencing. The result is 1: 1. Stable DJ-1 silencing MGC803 cells were successfully constructed. DJ-1 interference plasmids and negative control plasmids were transfected into MGC803 cells respectively. Western Blot,q RT-PCR was used to verify the interference effect of plasmid A to DJ-1. After transfection, Puromycin was used to screen. Establishment of stable DJ-1 silencing MGC803 cell line. 2. The effects of DADS and DADS on the proliferation and invasion of MGC803 cells. The results showed that both DJ-1 silencing and DADS could inhibit the proliferation of MGC803 cells. DJ-1 silencing can enhance the inhibitory effect of DADS on the proliferation of MGC803 cells. The results of scratch test and Transwell invasion test showed that the migration and invasion ability of MGC803 cells was decreased after DJ-1 silencing and DADS treatment (P0.05), and DJ-1 silencing enhanced the ability of DADS to inhibit tumor migration and invasion. 3. The DJ-1 expression of MGC803 cells was increased before and after dads treatment. The results of cytoimmunofluorescence Q RT-PCR,Western Blot assay showed that DJ-1 mainly distributed in cytoplasm of MGC803 cells treated with dads significantly decreased the expression of DJ-1,p-Akt (P0.05) .4.The silencing of DJ-1 and the effect of DADS on the tumorigenic ability of MGC803 cells in nude mice Tumorigenic assay in nude mice showed that DJ-1 silencing and DADS could significantly inhibit the tumorigenesis of MGC803 cells in nude mice. The immunohistochemical results showed that the positive expression of DJ-1 was significantly decreased and the expression of PTEN was significantly increased in the transplanted tumor tissue after silencing and DADS treatment. In addition, compared with the control group, the positive expression of Ki-67,CD-34,Vimentin was decreased and E-cadherin was significantly increased after DJ-1 silencing and DADS treatment. Conclusion: 1. Dads can down-regulate the inhibition of MGC803 cell proliferation and migration invasion by DJ-1 via PTEN/Akt pathway. 2. DJ-1 silencing can inhibit the proliferation and migration of MGC803 cells. Enhancing the inhibitory effect of DADS on proliferation and migration and invasion of MGC803 cells. 3. Both dads and silencing DJ-1 can inhibit the tumorigenesis of MGC803 cells of human gastric cancer, and both of them have synergistic effects.
【学位授予单位】:南华大学
【学位级别】:硕士
【学位授予年份】:2016
【分类号】:R735.2
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