| Prostate cancer is a common disease in elderly men.The incidence of prostate cancer ranks second among all malignant tumors and fifth among malignant tumors in the world.In 2016,the number of prostate cancer patients worldwide reached 1.4million,which is one of the major tumors endangering the health of men worldwide.In our country,the incidence of prostate cancer is increasing year by year due to the change of lifestyle and the aggravation of aging process,and most of the new prostate cancer patients in our country are locally advanced or widely metastasized.Androgen blocking therapy has become the standard treatment for these patients with advanced prostate cancer.Drugs used in ADT therapy include luteinizing hormone-releasing hormone agonists and androgen receptor antagonists.After 12 to 24 months of remission after ADT treatment,prostate cancer cells tend to gradually weaken their response to ADT treatment and eventually turn into castration-resistant prostate cancer.At present,radiotherapy or drug therapy for CRPC is not effective and specific,so exploring drugs that can selectively induce CRPC cell death has far-reaching impact on the treatment of prostate cancer.There are many ways of cell death,including apoptosis,autophagy and necrosis.At present,the research of anti-cancer drugs is mostly concentrated in the field of apoptosis,but almost all of the cancer cells will appear apoptotic resistance,which limits the application of this kind of drugs.Therefore,exploring non-apoptotic cell death mode has gradually become a research hotspot to improve the efficacy of anti-cancer treatment.In 2012,Dixon et al.studied the antineoplastic drugs for fibrosarcoma,and found that the compound erastin could induce tumor cells to produce a cell death mode regulated by cystine transport pathway,characterized by mitochondrial shrinkage,increased bilayer membrane density,iron-dependent lipid peroxidation,and named Ferroptosis.Recent studies have shown that erastin can induce Ferroptosis in tumor cells activated by RAS signaling pathway in various tumors,including renal cell carcinoma.Dixon et al.pointed out that the mode of cell death depends on the source,target and extent of oxidative damage.Therefore,exploring the effect factors and regulation mechanism of oxidative damage is the key to understand cell death including Ferroptosis.Because of the high activity of antioxidant system in cancer cells,it determines that cancer cells have a strong tolerance to oxidative stress.Among them,transcription factor NF-E2-related factor 2 is the core transcription factor that regulates oxidative stress in cells,plays a strong anti-oxidant/anti-apoptotic role,and is an important mechanism of drug resistance in cancer cells.When cells or organisms are exposed to reactive oxygen species,ROS can induce modification of cytoskeleton-related inhibitor protein Keap1(e.g.phosphorylation),which can dissociate the Keap1-Nrf2 complex,transfer Nrf2 from cytoplasm to nucleus,and bind to antioxidant response elements of antioxidant enzyme genes regulated by ROS,thus increasing the expression level of antioxidant protein and promoting cell survival.Heme oxygenase 1,?-glutamyl cysteine synthase(?-GCS)and quinone oxidoreductase 1(NQO1)are the main target proteins of Nrf2 in regulating oxidative stress.In order to further explore the mechanism of Nrf2/HO-1 signaling pathway regulating Ferroptosis in CRPC cells,we intend to use erastin-induced Ferroptosis model in CRPC cells,and use gene silencing technology to regulate the expression level of Nrf2/HO-1 to reveal the regulatory role of Nrf2/HO-1 in Ferroptosis of CRPC cells.PART ?OBJECTIVE: To study the changes of peroxidation level and Nrf2/HO-1 protein level during Ferroptosis in CRPC cells by MDA kit detection,flow cytometry and Western blot.METHODS: Based on the model of Ferroptosis induced by erastin in CRPC cells(PC3 ?DU145cell lines),the levels of lipid peroxidation and Nrf2/HO-1 protein during Ferroptosis of CRPC cells were studied by MDA kit,flow cytometry and Western blot.RESULTS: Compared with the control group,the ROS level in the experimental group was increased by flow cytometry.(2)The level of lipid peroxidation in the experimental group was increased by MDA kit.(3)Western blot showed that the levels of Nrf2 and HO-1 protein in CRPC cells treated with erastin increased.CONCLUSION: The levels of intracellular peroxidation and Nrf2 and HO-1 protein increased during Ferroptosis in CRPC cells.PART ?OBJECTIVE: To study the changes of peroxidation level and Nrf2/HO-1 protein level during Ferroptosis in CRPC cells by MDA kit detection,flow cytometry and Western blot.METHODS: Based on the model of Ferroptosis induced by erastin in CRPC cells(PC3 ?DU145 cell lines),the levels of lipid peroxidation and Nrf2/HO-1 protein during Ferroptosis of CRPC cells were studied by MDA kit,flow cytometry and Western blot.RESULTS: Compared with the control group,the ROS level in the experimental group was increased by flow cytometry.(2)The level of lipid peroxidation in the experimental group was increased by MDA kit.(3)Western blot showed that the levels of Nrf2 and HO-1 protein in CRPC cells treated with erastin increased.CONCLUSION: The levels of intracellular peroxidation and Nrf2 and HO-1 protein increased during Ferroptosis in CRPC cells.PART ?OBJECTIVE: To study the changes of peroxidation level and Nrf2/HO-1 protein level during Ferroptosis in CRPC cells by MDA kit detection,flow cytometry and Western blot.METHODS: Based on the model of Ferroptosis induced by erastin in CRPC cells(PC3 ?DU145 cell lines),the levels of lipid peroxidation and Nrf2/HO-1 protein during Ferroptosis of CRPC cells were studied by MDA kit,flow cytometry and Western blot.RESULTS: Compared with the control group,the ROS level in the experimental group was increased by flow cytometry.(2)The level of lipid peroxidation in the experimental group was increased by MDA kit.(3)Western blot showed that the levels of Nrf2 and HO-1 protein in CRPC cells treated with erastin increased.CONCLUSION: The levels of intracellular peroxidation and Nrf2 and HO-1 protein increased during Ferroptosis in CRPC cells. |
PDF Full Text Request