| Objective: Ovarian cancer is a serious threat malignant tumor to the women.It’s first mortality in the gynecology malignant tumor.The expression of activating transcriptional factor 4(ATF4)in tumor cells is higher than normal,and it’s can be increased by tumor micoenvironment signals(such as anoxic/oxidative stress and endoplasmic reticulum stress,etc).Some studies have found ATF4 involved in tumor growth,invasion and drug resistance of important signaling pathways.ATF4 can combine with VEGF-A,and u PA promoter,enhance its transcriptional activity to increase the expression.The studies suggested that VEGF-A and u PA in play an important role in the growth of ovarian cancer metastasis.Ribosomal protein L41(RPL41),Is a kind of small molecular peptides,is composed of 25 amino acid basic peptide,The present study shows that lower rpl41 is associated with a variety of tumor cells.Our team early research has shown that RPL41 can induce ATF4 phosphorylation,and transferred to the protease weight rapid degradation.And RPL41 molecular weight is small,can freely through the cell membrane,has the congenital advantage of drugs.This experiment using RPL41 induced ATF4 expression,to explore the inhibition effect of RPL41 for ovarian cancer cells and mechanism of action,aims to provide new targets for the treatment of ovarian cancer.Methods: 1.RPL41 can inhibit the growth of ovarian cancer cell line A2780 cells,CAOV3 cells,and cause ovarian cancer cell apoptosis morphological changes and influence on ovarian cancer cell cycle.(1)Using the Cell Titer-Glo ? fluorescent to detect cells activity with the treatment of RPL41.(2)Using Hoechst 33258 staining to observe cells apoptosis morphology(3)Using flow cytometry instrument to determinate the apoptosis of ovarian cancer cells.2.Wound healing assay and transwell cell invasion assays can research RPL41 effect on ovarian cancer cell migration ability.(1)Using the Wound healing assay can observe ovarian cancer cell migration ability with the treatment of RPL41.(2)Using the cell invasion assays can observe ovarian cancer cell invasion ability with the treatment of RPL41.3.Explore the molecular mechanisms of RPL41 inhibit the ovarian cancer cells growth,invasion and metastasis.(1)Using Luciferase reporter assay study ATF4 to readjustment of the promoter VEGF-A and u PA.(2).Using Western blot and RT-PCR to verify RPL41 can reduced ATF4 degradation,whether can down-regulated the expression of VEGFA and u PAResults: Hoechst33258 fluorescent staining and Annexin V/Pl double staining to detect apoptosis results showed that after the treatment of different concentration RPL4124 h,with the increase of RPL41 concentration,cell apoptosis rate are increased,When RPL41 concentrations greater than 100μM,difference statistically difference(p <0.05).However,after 48 h,And the trend of apoptosis is not clear,have no statistical difference.Cell Titer-Glo ? experimental results show that cell activity increased with the RPL41 increase of concentration,and the decrease of time.The results of Wound healing assay and transwell cell invasion assays show that : RPL41 can effectively inhibit ovarian cancer cell migration and invasion ability,statistically significant difference(p <0.01).Dual Luciferase reporter assay suggest that ATF4 can regulate the promoter of VEGF-A and u PA,.and Western blot and RT-PCR results show: ATF4,VEGF-A and uPA as RPL41 concentration increased,lower expression.Conclusion: 1 RPL4 induced ovarian cancer cells changes in cell morphology,inhibit the growth of cells,promote cell apoptosis,and acts on the G2-M phase cells.2.RPL41 inhibition of ovarian cancer cell migration and invasion ability.3.RPL41 induced ATF4down-regulation,and then regulate the VEGF-A and u PA promoter regulation.4.RPL41 can serve as a medium to down-regulation the ATF4... |
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