A Preliminary Study Combined AuNcs With MiRNA To The Gene Therapy Of Liver Cancer

Hepatocellular carcinoma (HCC) is one of the most common cancers in theworld, which has highly mortality and morbidity in recent years, especially in China.In the past few decades, traditional treatments of HCC have been improvedsignificantly, such as surgery, chemotherapy and radiotherapy, but the prognosis ofthese methods are still not optimistic. At the same time, the drawbacks of thesemethods are very obvious. Recently, with the rapid development of molecular biology,more and more people pay attention to the gene targeting treatment of liver cancer,which bring a lot of hope to cure cancers and other serious diseases.Objective:According to the gene and Au Nanocages together, observing the inhibition ofhuman hepatoma cell line SMMC-7721, and exploring the molecules and biologicmechanisms of the gene therapy combined with Au Nanocages simultaneously.Methods:According to the miRNA design software: miRWalk, the four23bpanti-miR-181were designed, and transfection is mediated by LipofectaminTM2000inliver cancer SMMC-7721cells. SRB method was applied to observe the effects ofanti-miR-181on cell growth; cell cycle phase change was detected by PI. In addition,cell apoptosis was detected by Annexin V-FITC/PI. Anti-miR-181b as therapeuticagents, mixed with1-tetradecanol.According to DSC to analyse the impact of1-tetradecanol with anti-miR-181b. In vitro studies, the stability and release propertiesof the mixture were observed by heating or ultrasonic. Whether the heating affects thecell growth was also tested. The selected PCM-anti-miR-181b was encapsulated inAuNCs, and encapsulation efficiency was calculated according to the relevantmathematical formula, After which, the inhibition of drug-loaded AuNCs, negativecontrol and blank AuNCs on SMMC-7721were observed by SRB.Results:Four anti-miR-181sequences were designed, and one effective sequences,anti-miR-181b, was selected; The SMMC-7721cells were transfected withanti-miR-181b, which induced cell apoptosis, and blocked the SMMC-7721cell cycledue to inhibition on the growth of hepatoma cells; The optimum mixing ratio ofPCM-anti-miR-181b was2:1, and1-tetradecanol did not change the nature of the anti-miR-181b, The complex could keep stable when it was stored for one month.Ofcourse, heating has no effect on the SMMC-7721cells, which can be used forhyperthermia; AuNCs can be used to deliver drugs, and1-tetradecanol occupies53%of the interior volume of AuNCsand the mole of anti-miR-181b encapsulated in eachAuNCs is3.06×10-25mol/cage.When they were heated to a temperature of40℃,anti-miR-181b obviously released from AuNCs, but it did not happen at roomtemperature and37℃. Cultured with anti-miR-181b-loaded AuNCs after24h,48hand72h, the cell viability were measured by the method of SRB,and the inhibition ofanti-miR-181b was30.457±1.56%、33.257±1.37%and27.490±1.19%, respectively atthe above time.Conelusions:we conclude that the AuNcs are promising gene delivery systems, which may becombined with gene therapy used to treat the liver cancer.