Co-loaded Cisplatin And Curcumin With Nano-liposomes For The Treatment Of Liver Cancer

Objective:The common treatment of cancer is surgery,radiotherapy,chemotherapy,targeted therapy and immunotherapy,and chemotherapy is one of the most important treatment.However,the toxicity of chemotherapy drugs and the drug resistance of the tumor are affecting the therapeutic effect of chemotherapy drugs,therefore,a variety of chemotherapy drugs are combined to treat cancer.In clinical treatment,the most common method is the combination of drugs acting on different targets or exerting different mechanisms.Compared with a single treatment with chemotherapeutic drugs,the combination of multiple drugs acting on different targets or with different mechanisms exerting a additive or even synergistic anti-cancer effect,while reducing drug dose,tolerance and side effects.Studies have shown that curcumin?CUR?can enhance the chemosensitivity of cytotoxic chemotherapy drugs?such as cisplatin,CDDP?by regulating a variety of signaling pathways.Therefore,the combination of CUR and CDDP will enhance the anti-cancer effect of CDDP.But the effect of drug combination in a large extent depends on the proportion of drugs and their effects in the process of the body,the time and proportion of each drug reaching the tumor site will be different due to the drug in vivo transport,metabolic processes are different,therefore,the treatment effect may be greatly reduced.In this subject,we try to be a certain proportion of CDDP and CUR were loaded into the liposome,to synchronize the two drugs in the body of the process,so that the two drugs can reach the tumor in the proportion of synergies,in order to achieve enhanced efficacy,reduce toxicity and prevent drug resistance.Methods:First,the optimal ratio of CDDP and CUR to hepatoma HepG2 cells was screened by median-effect method,and the reverse microemulsion method and membrane dispersion method were used to establish the nano-liposome?CCL?,along with the morphology,particle size,potential and in vitro release properties of CCL were investigated.Then,the uptake of CDDP and CUR in CCL was quantitatively analyzed in HepG2 cells,and the anti-tumor effect of CCL was evaluated by MTT method,Hoechst 33258 nuclear staining cells method and Annexin V-FITC/PI double staining method,and DCFH-DA fluorescent probe and Western blotting were used to study the molecular mechanism of CUR by increasing ROS in the cells to enhance the anti-cancer effect of CDDP.Finally,the pharmacokinetic properties of CCL in mice were studied by using Kunming mice as model,and the pharmacokinetics of CCL in vivo was studied by using mouse hepatoma H22 and human hepatocellular carcinoma HepG2 Evaluation.Results:The synergistic effect of CDDP and CUR on HepG2 cells in the ratio of1:8 was the best.The encapsulation rates of CDDP and CUR were 99.5%and 93.4%,respectively.The CCL potential was 10.8±2.5 mv and the particle size was about 100nm under TEM.In the solution of pH 7.4,the cumulative release of CDDP and CUR at 24h was 68.2%and 74.1%,respectively.CDDP and CUR in CCL reached the maximum uptake at 1 h on HepG2 cells,and then the intake was slowly reduced.The IC₅₀ of F-CDDP,CDDP-L,F-CUR,CUR-L,FCC and CCL was 5.6,2.56,20.24,20.33,1.18 and 0.62?M,respectively.The IC₅₀ of the combination group was significantly lower than that of the single drug administration group,that is,CDDP and CUR combination can produce stronger cell proliferation inhibition,and CCL group cell proliferation inhibition better than FCC.Nuclear staining cells and Annexin V-FITC/PI double staining once again showed that the combination of CDDP and CUR on HepG2 cells had the strongest inhibitory effect on proliferation,and the inhibitory effect was further enhanced by liposome loading.Mechanism studies show that CUR can activate p-ERK1/2 and suppress Sp1 by regulating ROS-related pathway,thus enhancing the inhibitory effect of CDDP on HepG2 cells.Pharmacokinetics experiments showed that CDDP and CUR in CCL compared with F-CDDP and F-CUR,AUC increased significantly,MRT and t_1/2z/2z were prolonged and CLz decreased.The pharmacodynamics of the two transplanted tumor models showed that CDDP and CUR combination could effectively inhibit the growth of tumor volume and prolong the survival time of mice,and the effects of them were further enhanced by liposome co-delivery,accompany by same weight gain with control,and the doses of CCL reduced by half could still be achieved considerable treatment effect with CDDP-L,indicating that CDDP and CUR co-loaded nano-liposomes can also reduce CDDP toxicity by reducing the doses of CDDP.Conclusion:The combination of CDDP and CUR can enhance the anti-tumor effect.The prepared cisplatin and curcumin co-loaded nano-liposomes?CCL?can further enhance the anti-tumor effect,and CCL can reduce the side effects of CDDP treatment by reducing doses of CDDP.