Design and Evaluation of Nanomedecine Targeting Cancer Stem Cells Therapy and Biosafety

Author:Li Bin

Supervisor:zhang zhi hong


Degree Year:2019





Objective: To determine the iron metabolism in cancer stem cells,and to screen the iron metabolism genes that play an important role in tumor initiation and the mechanism.To clarify the effect of graphene oxide on chemosensitivity treatment of prostate cancer,and to discover the effect of low concentration of graphene oxide on EMT of prostate cancer.Methods: We examined the iron metabolism in cancer stem cells and found that many genes related to iron regulation have undergone significant changes.According to the method of gene transcriptome sequencing,the abnormal expression of genes in our cancer stem cells and primitive tumor cells was studied.The main regulatory genes;we then found through relevant animal experiments that cancer stem cells have strong tumor-initiating ability,we constructed BMPR1 A,TFRC stable and low-expressing cell lines,and further tested tumor stem cell tumorigenic ability;The growth curve was recorded.The iron metabolism-regulated proteins in tumor tissues were detected by Western blotting PC3 cells were treated with graphene oxide at 10μg/ml,and the effects of graphene oxide exposure on cell migration,invasion and apoptosis were studied.Transwell experiment,flow cytometry,q-PCR and Western Blot were tested to study the effect of graphene oxide exposure on the expression of chemo-sensitive ABC protein family members in PC3 cells.Results: Tumor stem cells have strong tumor initiating ability and tumors.According to the methods of RT-qPCR and WB protein detection,the expression levels of TFRC,BMPR1 A,HFE and other proteins were significantly increased(P <0.05).We verified that tumor stem cells have strong tumor initiating ability and constructed by animal experiments.The low expression plasmids such as TFRC and BMPR1 A were established.After establishing the experimental model of tumor-bearing mice,the tumor formation ability of subcutaneous implanted tumor volume in the control group and the experimental group was significantly changed,and the results were statistically significant(P <0.05).According to the WB protein assay,graphene oxide can significantly reduce the expression of ABC protein family proteins and reduce the chemosensitivity of prostate cancer cells.Flow cytometry revealed that graphene oxide can significantly increase the apoptosis of prostate cancer cells.Conclusion: Tumor stem cells have a strong tumor-initiating ability.We found that there is a significant change in the homeostasis of iron metabolism in cancer stem cells.Among them,TFRC,BMPR1 A and other genes may play an important role in the maintenance of tumor stem cell iron homeostasis and tumor stem cell capacity.Graphene oxide can promote the efficacy of chemosensitivity treatment of prostate cancer by inhibiting the expression of ABC protein family members associated with chemosensitivity.Abnormal iron metabolism plays an important role in the maintenance of tumor stem cell capacity.Therefore,we synthesized DFCAF small molecule compounds and liposomes loaded with DFO,and verified that DFCAF has strong ability to inhibit tumor stem cells and EMT processes,and lipids.The plastid-loaded DFO can significantly reduce the iron load of cells and high-iron mice.In conclusion,this study evaluated the role of iron metabolism in the metabolism of cancer stem cells,and synthesized related molecular targeted drugs for the treatment of cancer stem cells.