Survivin Antisense Oligodeoxynucleotide Loaded Liposome Enhanced The Apoposis Of Hepatoma Carcinoma Cell Line Hepg2 Induced By Adriamycin

Objects Hepatocellular carcinoma(HCC) is one of the most common malignant tumors in our country, which is gravely threatening people's health because of its high malignant degree and bad prognosis. Exairesis is still the first-line and most effective treatment to HCC nowadays. Chemotherapy, as an important adjunctive treatment to HCC, has to settle the two tough difficulties of cytotoxicity to normal cells and drug resistance of tumor cells. It is researched that Survivin is the most powerful inhibiting apoptosis factor ever been found with the dual function of inhibiting apoptosis and regulating cellcycle, and it has a close relationship with chemoresistance. Nanoparticle gene transporter, an non-viral gene transporter,is well developed in recent years because of its safety, efficacy and fine property. In this paper:to use Survivin antisense oligodeoxynucleotide (ASODN) loaded liposome to enhance the apoposis of HCC induced by adriamycin(ADM).Methods To transfect the Survivin ASODN loaded liposome into hepatoma carcinoma cells, then to detect the change of Survivin by methods of RT-PCR and Western blot, to detect cytostasis using MTT assay, and to detect apoptosis rate of HCC using flow cytometry(FCM). After acting with ADM, to detect cytostasis using MTT assay and to observe the change of cell apoptotic index using FCM.Results 24 hours after all the groups having been transfected by Survivin ASODN, the mRNA and expression of proteins of them all decreased, especially the one of the group with the concentration of 800ng/ml of Survivin ASODN, and there was significant difference with the control group(P<0.05); MTT assay results showed that all cells group's proliferation activity degraded disparity after having been transfected by survivn ASODN. FCM detection results showed that 24 hours after the efficient of ASODN the apoptosis rates were 7.45±0.14%,13.15±0.37%, 35.75±0.68% respectively in the HepG2 groups with concentration of 400ng/ml, 600ng/ml and 800ng/ml, and there was significant difference contrasting to blank group with apoptosis rates of 0.54±0.05% and SODN transfected group with apoptosis rates of 0.56±0.08%(P<0.05). After the acting of ADM, apoptosis rate of HepG2 was 41.35+0.68% in the group with concentration of 600ng/ml,and there was significant difference contrasting to the apoptosis rates of 16.94+0.54% or 15.07+0.35% respectively in groups using ADM or ASODN transfected only.MTT assay results showed that the cytostasis degree had obvious difference (P<0.05) between blank control group with ADM group or ASODN group 24 hours after having been transfected.,and there was significant difference between ADM+ASODN group with each of other three groups (P<0.05).Conclusions The transfection induced by Survivin ASODN which has efficient function of blocking loaded liposome gene carrier which is safe and efficient has high transfection efficiency. Using liposome carrier to induce the transfection of Survivin ASODN, it can be more efficient to block survivin and down regulate the expressions of protein and mRNA of surviving which can promote the apoptosis of HepG2 finally. Using ASODN to block expression of survivin can inhibit the growth of HCC efficiently and increase the apoptosis of cell. Also, that can improve the chemosensibility of ADM and the effect of routine chemotherapy. All these make it potential in clinical application.