Humans ; Non-alcoholic Fatty Liver Disease

OBJECTIVETo explore the effect of microRNA-30a-5p (miRNA-30a-5p) on the biological behavior of human hepatoma cells.

METHODSThe liver cancer cell line SMCC-7721 cells and the normal liver cell line L02 cells (control) were transiently transfected with miRNA-30a-5p mimics and an miRNA-30a-5p inhibitor by Lipofectamine 2000 (Life Technologies). miR-30a-5p mRNA expression was detected by quantitative real-time (q)PCR. Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8) assay and apoptosis was assessed by flow cytometry.Invasion and migration were measured by transwell chamber assays. The SMCC-7721 cells was injected subcutaneously into nude mice to establish a tumor animal model.

RESULTSThe SMCC-7721 cells transfected with miRNA-30a-5p mimics showed significantly higher miRNA-30a-5p mRNA expression than the non-transfected SMCC-7721 cells and the transfected control L02 cells (P<0.01). The miRNA-30a-5p mRNA expression was significantly lower in the SMCC-7721 cells transfected with the miRNA-30a-5p inhibitor than the non-transfected SMCC-7721 cells the control L02 cells (P<0.01). The overexpression of miRNA-30a-5p inhibited the viability, colony formation rate, and invasion and migration abilities, as shown in the cells transfected with the miRNA-30a-5p mimics (P<0.05); in addition, the miRNA-30a-5p promoted proliferation of cells (P<0.05), as shown by more S phase cells detected by flow cytometry. SMCC-7122 cells transfected with miRNA-30a-5p mimics produced tumors with significantly higher average weight than tumors produced by SMCC-7122 cells that were untransfected or transfected with empty vector (both P<0.01).

CONCLUSIONOverexpression ofmiR-30a-5p had an inhibitory effect on cell proliferation, induced apoptosis, increased the number of cells in S phase, and markedly inhibited invasion and migration of SMCC-7721 HCC cells in vitro and in vivo.

Animals ; Apoptosis ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; Mice ; Mice, Nude ; MicroRNAs ; Neoplasm Invasiveness ; Transfection

Objective To prepare the Fe3O4-1oaded biodegradable liquid-solid phase inversion poly(lactic-co-glycolic acid) (PLGA) in situ implant for ultrasound-guided injection into nude mouse tumor model,and to investigate its clinical effect in thermomagnetic treatment of nude mice with human liver cancer SMMC-7721 cells in an alternating magnetic field.Methods An in situ implant containing 10％ Fe3O4 was prepared,and 50 μtl Fe3O4-PLGA-NMP gel was injected into the subcutaneous tissue of Kunming mice.The degradation of this material was observed for 2 consecutive months,and the changes in body weight were recorded.HE staining and Prussian blue staining were performed for the heart,liver,spleen,lung,and kidney of Kunming mice.Fresh ex vivo bovine liver was taken and cut into cubes with a dimension of 2 cmu×2 cm×2 cm and then 50 μl Fe3O4-PLGA-NMMP gel was injected;after phase inversion,the cubes ofex vivo bovine liver were heated for 1,2,3,4,and 5 minutes,respectively,and then cut open for observing the range of ablation;HE staining was also performed.Micro-CT scan was performed after ultrasound-guided injection of 50 μl Fe3O4-PLGA gel into the tumors of the nude mice,and then the nude mice were divided into treatment group and control group.The mice in the treatment group were given thermomagnetic treatment for 3 minutes,and tumor growth was observed daily.Results The biodegradation of Fe3O4-PLGA-NMMP implant showed that the subcutaneously injected material was gradually metabolized at 2 weeks after injection and that the nude mice were in good condition.The bovine liver ablation experiment showed that the range of ablation of 50 μl Fe3O4-PLGA implant reached 1.46 ± 0.11 cm.HE staining showed that part of bovine liver had coagulative necrosis.The phase inversion experiment of Fe3O4-PLGA gel showed quick liquid-solid phase inversion of the material after injection into the tumor,and the process of liquid-solid phase inversion could be monitored by ultrasound and CT.The detachment and incrustation of the tumor started at 2 days after treatment,the wound started to heal 15 days later,and the tumor tissue disappeared completely.Conclusion Ultrasound-guided injection of biodegradable Fe3O4-PLGA in situ implant combined with magnetic thermal ablation can effecfively treat human liver cancer SMMC-7721 cells in nude mice.