| More and more literatures have reported that the incidence of both type 2 diabetes and malignant tumor cases are rising, lots of malignant tumor patients are also suffering from type 2 diabetes. According to latest statistics, 47% of liver cancer patients have diabetes, and 16% breast cancer patients have diabetes. It is found that type 2 diabetes can increase the risk of liver cancer, breast cancer and other malignant tumor which is considered highly correlated with malignant tumor. The latest retrospective analysis found that, compared with patients without diabetes, cancer patients with diabetes have worse chemotherapy effects, higher recurrence and mortality rate. Although the relationship between diabetes and cancer has been attracted attention in clinical, the reasons and mechanisms of diabetes causing tumor resistance, influencing tumor recurrence rate and mortality are still poorly understood. It is short of effective interventions to reduce the recurrence rate of tumor with disease and increase the survival rate of patients with diabetes. Therefore, in view of liver cancer and breast cancer with high incidence and mortality rate causing by diabetes(hyperglycaemia and hyperinsulinemia), which are modeled after the chemotherapy sensitivity that the hyperglycemia and hyperinsulinemia will infuence liver cancer and breast cancer. Study the effect of diabetes on liver cancer and breast cancer chemotherapy as well as the mechanism, design and synthesis a series of organoseleniums to synergy the chemotherapy sensitization of first-lines of defense against tumors. Thus, we can improve the cure rate of liver cancer and breast cancer patients with diabetes effectively. The scratch test, tumour invasion assay were used to evaluate whether hyperglycemia and hyperinsulinemia can promote tumor cell proliferation in vitro; MTT assay was used to test the ability of organoselenium and/or doxorubicin, selenium nanoparticles stimulating the tumor cell proliferation under different glucose and insulin concentration. Choosing phenylbenzo [1,2,5] selenadiazole derivatives with different substituents to explore the cell signal harass and magnified in the process of apoptosis which was induced by anti-tumor drugs; Western blotting technique was used to discuss the potential mechanisms of phenylbenzo [1,2,5] selenadiazole derivatives enhance the first-line chemotherapy drugs effect under the condition of hyperglycemia; The confocal was used to test whether selenium nanoparticles can stimulate the occurrence of autophagy, to provide the scientific basis to broaden the design idea of organoselenium and selenium nanoparticles, develop more cleary targets and mechanisms of specific new low-toxic diabetes associated with liver cancer chemotherapy drugs or novel leading compounds. The main results were as follows:1. Chapter 2 study whether phenylbenzo [1,2,5] selenadiazole derivatives can reverse chemotherapy resistance of HepG2 cells to doxorubicin induced by hyperglycemia and discuss the involved mechanisms. This study found that hyperglycemia can stimulate the proliferation of HepG2 cells, reduce chemotherapy sensitivity to doxorubicin while phenylbenzo [1,2,5] selenadiazole derivatives 2a can induce HepG2 cells apoptosis by improving the intracellular uptake ability to doxorubicin, causing more ROS accumulation, DNA damage and inducing mitochondria fracture, which can antagonize the drug-resistant induced by hyperglycemia. To illuminate the sensitization mechanism, we found that 2a could targeted the AMPK and p53 protein kinase, activated AMPK and p53 signaling pathway to antagonize the the drug-resistant induced by hyperglycemia.2. Chapter 3 study whether phenylbenzo [1,2,5] selenadiazole derivatives can reverse chemotherapy resistance of MCF-7 cells to doxorubicin induced by hyperglycemia and discuss the involved mechanisms. This study found that hyperglycemia can stimulate the proliferation, migration and invasion of MCF-7 cells and reduce chemotherapy sensitivity to doxorubicin, while phenylbenzo [1,2,5] selenadiazole derivatives 2c can induce MCF-7 cells apoptosis by improving the intracellular uptake ability to doxorubicin, causing more ROS accumulation, DNA damage and inducing mitochondria fracture, which can antagonize the drug-resistant induced by hyperglycemia. Western blotting method was used to clarify the sensitization mechanisms, we found that all the phenylbenzo [1,2,5] selenadiazole derivatives we synergized can activate AMPK protein. For further study through a series experiments,we found that 2c also can target p53 protein kinase, finally activated related kinases(p53, Akt, ERK, protein, etc.) and the Bcl-2 family, AMPK pathway(AMPK, p-AMPK, mTORC1, p-mTORC1, p70S6K1, p-p70S6K1, 4E-BP1, p4E-BP1), to evaluate whether the organoselenium can improve the chemotherapy resistance to doxorubicin on breast cancer cells.3. This chapter study whether the selenium nanoparticles can induce HepG2 cells apoptosis in hyperinsulinemia by incresing the autophagy ability and the involved mechanisms. In this study, we found that hyperinsulinemia can promote tumor cell proliferation, migration and invasion, and the study also found that hyperinsulinemia can reduce liver cancer cells autophagy ability. And hyperinsulinemia can inhibit tumor autophagy occurring, we suspected that hyperinsulinemia would decrease the chemotherapy effect. MTT method was used to test antitumor ability of SeNPs, uPA-CS-SeNPs 、 ACPP-CS-SeNPs and uPA/ACPP-CS-SeNPs. Compared with SeNPs-CS, uPA/ACPP-CS-SeNPs can significantly inhibit the proliferation of tumor cells. Flow cytometry instrument analysis found that they can lead to Hep G2 cells apoptosis. The Selenium nanoparticles linked up with two targets uPA and ACPP which has higher selectivity to cancer cells than SeNPs-CS and the antitumor effect is better. Further experiment represented by uPA/ACPP-CS-SeNPs, through the MDC experiment and mRFP-GFP-LC3 double fluorescent probe experiment, we found that hyperinsulinemia can reduce the HepG2 cells autophagy level, while uPA/ACPP-CS-SeNPs can increase the occurrence level of autophagy. Those results suggested that targeted selenium nanoparticles can selectively kill HepG2 cells may has relationship with autophagy occuring, which can antagonize high insulin induced chemo-resistance in tumor cells.In general, this paper verified two pathologic features of diabetes including hyperglycemia and hyperinsulinemia, both of them can reduce tumor chemotherapy effect. Various of biological methods were used to study the mechanisms of seven phenylbenzo [1,2,5] selenadiazole derivatives sensitized the chemotherapy effects to doxorubicin under hyperglycemia. The results suggested that the phenylbenzo [1,2,5] selenadiazole derivatives has huge potential application value in reverse tumor drug resistance. We also modified the selenium nanoparticles aiming at improving the compounds’ selectivity. We found that the double targeting SeNPs can induce tumor cell apoptosis by increasing the occurrence of tumor cell autophagy, antagonize cell autophagy ability in the hyperinsulinemia condition. Then the relationship between autophagy and apoptosis has been discussed. This paper widen the design ideas of antitumor organoselenium compounds and selenium nanoparticles, and provided the scientific basis more clearly, new low-toxic anticancer associated with diabetes drugs which has more clearly mechanisms. |
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