Experimental Research Of MAGL-shRNA Carried By Ultrasound Microbubble On Hepatocellular Carcinoma In Rats

Objective To research the effect after the transfection ofMonoacylglycerol lipase shRNA by using ultrasound-targeted microbubbledestruction on rats’hepatocellular carcinoma.Methods CBRH7919rats’ hepatoma cells were cultivated with1640supplemented with10%fetal bovine serum.Then,these cells wereimplanted subcutaneously in nude mice for about two weeks.Finely,we cutthe tumors into small organizations about1mm3and transplant theseorganizations into liver lobe of rats. The successful establishment ofmodels were testified by AFP and two-dimensional ultrosound.50ratswere randomly divided into4groups: phosphate-buffered solution group,lipid microbubbles loaded MAGL-shRNA (MAGL-shRNA+MB group),pure lipid microbubbles+ultrasound (MB+US group), and lipidmicrobubbles loaded MAGL-shRNA+ultrasound (MAGL-shRNA+MB+US group). Rats of each group were injected with1mlcorresponding reagents through the tail vein. The expression of MAGLprotein was detected by Western-blot and immunohistochemistry(IHC). MAGL mRNA levels were determined by2807TaqMan quantitativepolymerase chain reaction (qPCR) assay. Tumor size and tumor metastasisrate of rats were compared.Results Tissue of rat liver cancer was offwhite, its section is fish likechange, HE staining accords with the pathological manifestations ofprimary liver cancer pathology. The protein expression of MAGL in HCCtissue was higher than in normal tissue. Detection byImmunohistochemistry and Western Blot show MAGL expression in ratliver tissue was obviously higher than its expression in normal liver tissue;liver tissue protein,expression trend of each groups,protein expression ofMAGL-shRNA+MB group decreased significently.Expression ofMAGL-mRNA in MAGL-shRNA+MB+US group was lower than otherssignificently(P<0.05). Tumor size of MAGL-shRNA+MB+US group wassmaller than others(P<0.05). Tumor metastasis was found in all groups,tumor metastasis rate was lowest in MAGL-shRNA+MB+US group.Conclusions MAGL-shRNA transfected into liver tumor of rats couldinhibits the growth and the metastasis of tumors. Ultrasound targetedmicrobubble destruction can effectively transfect MAGL-shRNA into targettissues, which enhanced the effect of gene therapy in liver cancer.