| Background and objective:Liver cancer is one of the most common malignant tumors.At present,the most important clinical treatment is surgical resection.And there is still no effective treatment for advanced liver cancer.Traditional chemotherapeutic drugs do not have the targeting of liver cancer tissues,even often cause serious side effects.As a result,some patients gave up treatment,which greatly affected the treatment effect and prognosis.On the other hand,in the use of a carrier for drug delivery,artificially synthesized materials such as nano gold,liposome have immunogenicity and certain toxicity.In vivo,it can cause the body,s inflammatory response and decline the drug efficiency.In this study,peptides were binding with exosomes to enable them to target liver cancer tissues.Many researches show that microRNA is involved in many life activities,and miR-125b plays an important role in inhibiting the occurrence and development of liver cancer.We used miR-125b as the therapeutic drug and modified exosome as a earrier to treat hepatoeellular eancer.It is expected to make breakthroughs in the development of new treatment strategies for liver cancer.Experimental methods:1.The expression of miR-125b in serum exosomes of normal liver cancer patients and normal human serum exosomes was searched in the database,and analyzed the correlation with liver cancer;The real-time PCR was applied to test the expression of miR-125b in liver cancer cell line;We used miR-125b mimic to transfect liver cancer cells.Then CCK-8 kit,wound healing assay and transwell migration assy were applied to test the proliferation,migration of liver cancer cell lines.Westrn blot was used to examine the change of epithelial-mesenchymal transition related proteins.2.The expression of GPC-3 was detected by pathological histochemistry in clinical specimens.The expression of GPC-3 protein in liver cancer cell lines was detected by Western blot.In vitro we examined the affinity between L5(amino acid sequence RLNVGGTYFLTTRQ)and liver cancer lines.3.Collecting and identifying exosomes that secreted by mouse embryonic fibroblasts;L5 isbound to exosomes by click chemistry;Synthesis efficiency was detected by fluorescence microscopy;miR-125b was packed into exosomes by electroporation.4.Mouse hepatocellular carcinoma in situ model was established,and the distribution of above-mentioned exosomes were detected in invo by using the small animal living imaging device;The progression of the liver cancer in situ model was analyzed after the tumor-bearing mice were treated with above-mentioned exosomes by tail intravenous injection.Results:1.The expression of miR-125b in serun exosomes of patients with liver cancer was decreased,and it was negatively correlated with the prognosis of liver cancer(P<0.01).After transfection with miR-125b mimic in liver cancer cell lines,EMT-related protein showed interstitial-epithelial transformation(MET);wound healing assay and Transwell migration experiments demonstrated a weakened tumor migration capacity.2.L5 has a targeting property for GPC-3 high expression of liver cancer cells.3.L5 enabled exosomes to target liver cancer cells;miR-125b mimic can inhibit tumor migration through exosomes entering cells.4.Tumor-bearing mice which injected with L5-linked exosomes,compared to the common exosomes,had a better therapeutic effect in inhibition of tumor.Conclusions:The exosomes obtain the ability to target GPC?3 high expression after lined to L5;miR-125b has an inhibitory effect on the migration and growth of hepatocellular carcinoma;the engineered exosomes can increase the accumulation of drugs in liver cancer tissues.There are significant prospects for cancer treatment. |
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