[Optimization of preparation technology for Acanthopanax senticosus total saponins microemulsion by D-optimal mixture design and intestinal absorption characteristics].

Following the preparation of Acanthopanax senticosus total saponins microemulsion, the formulation and preparation technology were optimized and the quality was evaluated. The absorption characteristics of A. senticosus total saponins microemulsion by the self-microemulsifying drug delivery system(SMEDDS) were investigated in the unidirectional intestinal perfusion model in vivo. The oil phase, mass ratio(K_m), number of revolutions, and drug concentration were subjected to single-factor investigation with the area of pseudo-ternary phase diagram as the index. The process was optimized by D-optimal mixture design with the particle size as the index, and then the appearance, morphology, and particle size were investigated. The mass concentrations of eleutherosides B and E in the microemulsion were determined. The results showed that the optimum formulation of A. senticosus total saponins microemulsion was determined as follows: 20.8% of water phase, 31.2% of isopropyl palmitate, and 48.0% of soybean phospholipid and absolute ethanol(K_m=1∶1). As revealed by the observation under a transmission electron microscope, the microemulsion exhibited homogeneous dispersion and was a spherical emulsion droplet in the water-in-oil type. At room temperature, the pH value was 5.19, the refractive index 1.416 5, the average particle size(26.47±0.04)nm, and the polydispersity index(PDI) 0.118±0.03. The content of the eleutherosides B and E was 0.038 9 and 0.166 4 mg·mL~(-1), respectively. The preliminary stability study showed that the solution was clear and transparent within 30 d, without stratification or content change, indicating good stability. The absorption of microemulsion in each intestinal segment was significantly improved as compared with that of the A. senticosus total saponins, with the best absorption effect detected in the ileum, which has laid a foundation for further development and utilization of A. senticosus.

Adsorption and desorption characteristics of ginsenosides from Panax ginseng C. A. Meyer on middle-pressure chromatogram isolated gels.

Four types of middle-pressure chromatogram isolated gels are evaluated for adsorption or desorption characteristics of ginsenosides from Panax ginseng. Among them, SP207SS and SP2MGS were selected for dynamic investigations based on their static adsorption or desorption capacity of total ginsenoside. Their adsorption kinetics was better explained by pseudosecond-order model and isotherms were preferably fitted to Langmuir model. Dynamic breakthrough experiments indicated an optimum sample loading speed of 4 bed volume/h for either SP207SS or SP2MGS. Desorption speed was determined to be 2 bed volume/h according to desorption amount of total ginsenoside in their effluents. Eight ginsenosides were identified and quantified by high performance liquid chromatography-triple quadropole-mass spectrometry in total ginsenoside extract and different fractions during stepwise dynamic elution. For SP207SS, 27.62% of loaded ginsenosides was detected in 40% ethanol fraction, while 59.12% of them were found in 60% ethanol fraction. As on SP2MGS, the number went to 53.71 and 44.43%, respectively. Recovery rate of ginsenosides were calculated to 78.65% for SP207SS and 89.53% for SP2MGS, respectively. Intriguingly, content of Rg1 and Re in 40% ethanol fraction from SP207SS became 20.1 and 18.6 times higher than that in total ginsenoside extract by one-step elution, which could be leveraged for the facile enrichment of these two ginsenosides from natural sources.

Anti-Tumor Mechanisms Associated With Regulation of Non-Coding RNA by Active Ingredients of Chinese Medicine: A Review.

Cancer has become the second leading cause of death worldwide; however, its complex pathogenesis remains largely unclear. Previous research has shown that cancer development and progression are closely associated with various non-coding RNAs, including long non-coding RNAs and microRNAs, which regulate gene expression. Target gene abnormalities are regulated and engaged in the complex mechanism underlying tumor formation, thereby controlling apoptosis, invasion, and migration of tumor cells and providing potentially effective targets for the treatment of malignant tumors. Chemotherapy is a commonly used therapeutic strategy for cancer; however, its effectiveness is limited by general toxicity and tumor cell drug resistance. Therefore, increasing attention has been paid to developing new cancer treatment modalities using traditional Chinese medicines, which exert regulatory effects on multiple components, targets, and pathways. Several active ingredients in Chinese medicine, including ginsenoside, baicalin, and matrine have been found to regulate ncRNA expression levels, thus, exerting anti-tumor effects. This review summarizes the scientific progress made regarding the anti-tumor mechanisms elicited by various active ingredients of Chinese medicine in regulating non-coding RNAs, to provide a theoretical foundation for treating tumors using traditional Chinese medicine.

Three New Triterpenoid Saponins from Notoginseng Medicinal Fungal Substance.

Three new triterpenoid saponins, named ginsenoside-Rh23 (1), ginsenoside-Rh24 (2), and ginsenoside-Rh25 (3), were isolated from notoginseng medicinal fungal substance. Their structures were elucidated by a combination of 1D- and 2D-NMR, MS and chemical analysis. Compounds 1 - 3 exhibited moderate cytotoxic activity against MCF-7 and NCI-H460 cancer cell lines.

[Analysis of spatial distribution of artemisinin in Artemisia annua in China].

Geo-herbals are preferred by long-term clinical application of Chinese medicine, they are produced in a specific area and affected by the specific production and processing methods. Geo-herbals have a high reputation, compared with other regions produced by the same kind of herbs with good quality, efficacy and stable quality characteristics. Geo-herbal is a unique concept that has been formed by the history of Chinese civilization for thousands of years and the history of Chinese medicine. According to the definition of geo-herbal, it can be seen that there are some differences in the quality and efficacy of the same kind of medicinal herbs in different regions, and there is some similarity in the specific area. In this study, the spatial distribution characteristics of artemisinin in Artemisia annua were analyzed by spatial statistical analysis. ①The spatial autocorrelation of artemisinin content was analyzed by "I coefficient". The results showed that the content of artemisinin in each province was significant and positive spatial autocorrelation. The content of artemisinin in each province was not randomly distributed, but with a clear spatial aggregation characteristics. ②The spatial variability of artemisinin content was analyzed by "G statistic". The results showed that the area with high content of artemisinin was distributed in the southwest region, and the content of artemisinin was concentrated in the space, which tends to accumulate at high value. The content of artemisinin in Tianjin and Liaoning province is low, and it tends to accumulate at low value, while the content of artemisinin in other provinces is the transition zone of high and low value. ③Based on the index of artemisinin in each province, the spatial correlation of artemisinin content in each region was analyzed by "moran scatter plot". The results showed that the content of artemisinin in nine provinces such as Guangxi and Chongqing belonged to high value gathering area. ④Based on the artemisinin content and ecological environment factor of sampling point, the "geophysical detector" was used to analyze the continuous quantitative data and discontinuous qualitative data. The results showed that the sunshine, temperature and precipitation were the main factors affecting the content of artemisinin.

[Spatial differences of Chinese material medicine resources species in Jilin province].

The differences in the spatial distribution of regional Chinese material medicine resources between regions are determined by differences in natural conditions and social resources among different regions. Spatiotemporal evolution of the distribution of Chinese material medicine resources is a manifestation of the development process of Chinese medicine industry in time and space. The spatiotemporal evolution of the distribution of regional Chinese material medicine resources can reveal the regularity of spatial distribution of Chinese material medicine resources. County as the basic unit of the national material medicine resources census, the difference of resource endowment is of great significance to the rational development of Chinese material medicine resources policy and the coordinated development of regional resources and environment. Based on the results of the pilot project of Jilin province, this study selected 51 counties that had completed the census of Chinese material medicine resources. According to the species of Chinese material medicine resources, we explored the diversity of Chinese material medicine resources in Jilin province by using statistical data analysis (ESDA), trend surface analysis and spatial variability function to analyze the spatial differences of Chinese material medicine resources. Chinese material medicine resources are distributed in the eastern and southeastern part of Jilin province, mostly in the Changbai Mountains. The species of Chinese material medicine resources tend to be low-value spatial aggregation; the overall cold spots are located in Changchun and Jilin city, near the administrative center, mostly urban built-up area. The rich areas of Chinese material medicine resources are rich in hot spots, mainly in broad-leaved forest. The low-abundance of Chinese material medicine resources is dominated by cultivated vegetation.

[Research on extraction of ginseng planting distribution information based on object-oriented classification--by case study of Fusong country in Jilin].

Due to the particularity of ginseng cultivation, the soil fertility of cultivated ginseng is seriously depleted, so that the cultivated ginseng land can not be reused in the short term, and the land area available for cultivating ginseng becomes less and less with the growth of ginseng cultivation time. Therefore, in order to effectively manage ginseng cultivation, and achieve the sustainable use of ginseng land, it is necessary to obtain accurate information on the distribution of ginseng planting space. In this study, the object-oriented classification method based on rule set was used to extract ginseng planting area based on the ZY-3 satellite data in Fusong county, Jilin province. Firstly, multi-scale segmentation of ZY-3 remote sensing image in the study area was made, and the optimal segmentation scale was determined on the basis of multi-scale segmentation results. Secondly, a spectral curve according to the different feature type samples was generated. The similarities and differences between ginseng plot and other types of surface features were analyzed, and a rule set based on the results of spectral analysis was established to achieve the final extraction. The results show that the object-oriented classification method based on rule set can effectively extract the ginseng planting plots in the study area, and solve the problem that the extraction result is broken compared with the traditional pixel-based classification method.

Two new dammarane-type triterpenoid saponins from ginseng medicinal fungal substance.

Two new triterpenoid saponins, namely ginsenoside Re8 (1) and notoginsenoside ST-8 (2), were isolated from ginseng medicinal fungal substance. Their structures were elucidated by spectroscopic data and chemical analysis.

Two new dammarane-type triterpenoid saponins from notoginseng medicinal fungal substance.

Two new dammarane-type triterpenoid saponins, namely ginsenoside Rk6 (1) and ginsenoside-Rh22 (2), were isolated from notoginseng medicinal fungal substance. The structures of 1 and 2 were established as 3ß,6α,12ß,26-tetrahydroxydammar-20(21),24(25)(E)-diene-6-O-ß-D-glucopyranoside and 3ß,6α, 20(S)-trihydroxy-12(R),23(R)-expoxy-13(S),17(S)-dammar-24-ene-6-O-ß-D-glucopyranoside on the basis of spectroscopic analysis and chemical analysis, respectively.

Investigation of the anti-glioma activity of Oviductus ranae protein hydrolysate.

Oviductus Ranae is the dry oviducts of Rana temporaria chensinensis, and it has been reported to have a range of biological activities. This study aimed to investigate the effects of Oviductus Ranae protein hydrolysate (ORPH) on human glioma C6 cell proliferation and apoptosis in vitro and in vivo. Following in vitro treatment, cell viability and colony formation assays showed that ORPH inhibited C6 cell proliferation. In addition, the results of western blotting also demonstrated that ORPH effectively regulated the expression of the apoptosis related proteins, cleaved caspase-3, Bax and Bcl-2, DNA staining and flow cytometry analysis demonstrated that ORPH significantly promoted apoptosis in this cell line, a finding that was confirmed in vivo using terminal deoxynucleotidyl transferase dUTP nick end labeling. Further investigation demonstrated that ORPH increased apoptosis by modulating the release of inflammatory cytokines and the phosphoinositide 3-kinase (PI3K)/AKT signaling pathway; this was demonstrated using a PI3K/AKT inhibitor (NVP-BEZ235). In summary, the present study suggested that ORPH promoted apoptosis and inhibited glioma cell proliferation by influencing the PI3K/AKT signaling pathway.

Nontargeted metabolomic analysis and "commercial-homophyletic" comparison-induced biomarkers verification for the systematic chemical differentiation of five different parts of Panax ginseng.

A key segment in authentication of herbal medicines is the establishment of robust biomarkers that embody the intrinsic metabolites difference independent of the growing environment or processing technics. We present a strategy by nontargeted metabolomics and "Commercial-homophyletic" comparison-induced biomarkers verification with new bioinformatic vehicles, to improve the efficiency and reliability in authentication of herbal medicines. The chemical differentiation of five different parts (root, leaf, flower bud, berry, and seed) of Panax ginseng was illustrated as a case study. First, an optimized ultra-performance liquid chromatography/quadrupole time-of-flight-MS(E) (UPLC/QTOF-MS(E)) approach was established for global metabolites profiling. Second, UNIFI™ combined with search of an in-house library was employed to automatically characterize the metabolites. Third, pattern recognition multivariate statistical analysis of the MS(E) data of different parts of commercial and homophyletic samples were separately performed to explore potential biomarkers. Fourth, potential biomarkers deduced from commercial and homophyletic root and leaf samples were cross-compared to infer robust biomarkers. Fifth, discriminating models by artificial neutral network (ANN) were established to identify different parts of P. ginseng. Consequently, 164 compounds were characterized, and 11 robust biomarkers enabling the differentiation among root, leaf, flower bud, and berry, were discovered by removing those structurally unstable and possibly processing-related ones. The ANN models using the robust biomarkers managed to exactly discriminate four different parts and root adulterant with leaf as well. Conclusively, biomarkers verification using homophyletic samples conduces to the discovery of robust biomarkers. The integrated strategy facilitates authentication of herbal medicines in a more efficient and more intelligent manner.

Flavonoids Extraction from Propolis Attenuates Pathological Cardiac Hypertrophy through PI3K/AKT Signaling Pathway.

Propolis, a traditional medicine, has been widely used for a thousand years as an anti-inflammatory and antioxidant drug. The flavonoid fraction is the main active component of propolis, which possesses a wide range of biological activities, including activities related to heart disease. However, the role of the flavonoids extraction from propolis (FP) in heart disease remains unknown. This study shows that FP could attenuate ISO-induced pathological cardiac hypertrophy (PCH) and heart failure in mice. The effect of the two fetal cardiac genes, atrial natriuretic factor (ANF) and ß-myosin heavy chain (ß-MHC), on PCH was reversed by FP. Echocardiography analysis revealed cardiac ventricular dilation and contractile dysfunction in ISO-treated mice. This finding is consistent with the increased heart weight and cardiac ANF protein levels, massive replacement fibrosis, and myocardial apoptosis. However, pretreatment of mice with FP could attenuate cardiac dysfunction and hypertrophy in vivo. Furthermore, the cardiac protection of FP was suppressed by the pan-PI3K inhibitor wortmannin. FP is a novel cardioprotective agent that can attenuate adverse cardiac dysfunction, hypertrophy, and associated disorder, such as fibrosis. The effects may be closely correlated with PI3K/AKT signaling. FP may be clinically used to inhibit PCH progression and heart failure.

Two new triterpenoid saponins from ginseng medicinal fungal substance.

Two new dammarane-type triterpenoid saponins, namely ginsenosides Rb4 (1) and Rb5 (2), were isolated from ginseng medicinal fungal substance. The structures of 1 and 2 were established as 3ß,12ß,20(S)-trihydroxydammar-24(25)-ene-3-O-[α-d-glucopyranosyl-(1→4)-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl]-20-O-ß-d-glucopyranoside and 3ß,12ß,20(S)-trihydroxydammar-24(25)-ene-3-O-[α-d-glucopyranosyl-(1→4)-α-d-glucopyranosyl-(1→4)-ß-d-glucopyranosyl-(1→2)-ß-d-glucopyranosyl]-20-O-ß-d-glucopyranoside on the basis of spectroscopic analysis and chemical analysis, respectively.

Angelica sinensis reduced Aß-induced memory impairment in rats.

BACKGROUND: Studies have shown that Angelica sinensis (JiLin AoDong Medicine Industry Groups Co., Ltd., Jilin, China) root (AS) ameliorates various diseases, although its effects in Alzheimer's disease (AD) have not been elucidated. PURPOSE: The present study examined the effects of AS in a rat model of AD. METHODS: Positional Aß injections were administered to rats. The behavioral effects of AS administration were examined using the Morris water maze, and the molecular effects on gene and protein expression, and apoptosis, were determined. RESULTS: AS reversed the social behavioral impairments observed in this rat model of Aß-induced memory impairment. Western blot analysis also revealed lower hippocampal levels of Aß and ß-site amyloid precursor protein-cleaving enzyme. Terminal deoxynucleotidyl transferased UTP nick end labeling indicated that AS significantly inhibited apoptosis via effects on nuclear factor kappa B (NF-κB) signaling. Real-time PCR, enzyme-linked immunosorbent assay, and immunohistochemical staining indicated that AS effectively inhibited inflammation and upregulated expression of glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) in the hippocampus of this rat AD model. DISCUSSION: AS effectively rescued the symptoms of AD in a rat model by inhibiting inflammation, apoptosis, and NF-κB signaling pathway. CONCLUSION: These findings suggested that AS could provide a potential drug for the treatment of AD.

[Exploring pharmacological principle of Artemisia carvifolia with textmining technology].

To explore the pharmacological principle of Artemisia carvifolia,the text mining technique was used. All the references of A. carvifolia were collected from PubMed database, and then the rules of the main ingredient,relative diseases, organs, tissues, proteins and metabolites were analyzed. Finally, a network was set up. Then it was found that the main ingredients included sesquiterpenoids,flavonoids,and volatileoils.The diseases such as malaria, cerebral malaria, falciparum malaria, visceral leishmaniasis and systemic lupus erythematosus were often treated with A. carvifolia. In association in organ were the liver, skin, trachea,lungs,and spleen.Correlations with tissues were mainly including macrophages, T lymphocytes, blood vessels, epithelial cells.The protein was correlation with it involved CYP450, PI3K, TNF-α, AASDPPT, DNA polymerase and so on. Comprehensive and systematic treatment principle of A. carvifolia was obtained by text mining, which was helpful in clinical application.

[Review on study of Dao-di herbs Artemisiae Annuae Herba].

Artemisiae Annuae Herba has used as a medicine for more than 2 000 years. To infer based on the modern study results, Artemisiae Annuae Herba used for the treatment of malaria recorded in Zhou Hou Bei Ji Fang before 1 700 years should come from Artemisia annua. Based on the data of Chinese materia medica, from the field of treatment hotness and preventing attack of malaria etc., the Dao-di producing district of Artemisiae Annuae Herba should at Jingzhou (now Hubei) and surrounding areas in history. From the view of anti-malaria components artemisinin content, the Dao-di growing producing district of Artemisiae Annuae Herba should locate at Chongqing, Guangxi and its surrounding provinces. The study results showed that A. annua was harvested in flower bloom at autumn, and in this time it also had higher artemisinin content. If A. annua was stored exceed six months, artemisinin could be degraded about thirty percent. So it should be stored in a cool and dry place generally. Wild A. annua had a rich genetic diversity. Artemisinin content of A. annua breeding in experimental field could reache to two percent.

[Anti-tumor mechanism of Artemisia annua based on bioinformatics].

This paper was aimed to explore the relationship between Artemisia annua and tumor, and investigate its anti-tumor mechanism based on the bioinformatics molecular network analysis, text mining technique and other related methods. Text mining tool Polysearch database was used to get the information, and after formatting conversion, the information was imported into the bioinformatics analysis software Cytoscape3.2.1 for visualization and subsequent bioinformatics molecular network analysis .It was shown that the A. annua was associated with the tumor by 8 key proteins： TNF,VEGF,PI3K, ALDH1, Bcl-2, MicroRNA, p38 and CASP3 by text mining technique. The main biological processes involved in its anti-tumor effect included cell cycle, post-translational protein modification, cell cycle regulation, protein ubiquitination and organelle tissues regulation. The key network analysis showed that the action may be achieved by regulation of triglyceride metabolic process, positive regulation of triglyceride metabolic process, positive regulation of triglyceride catabolic process, regulation of budding cell apical bud growth, negative regulation of mitotic cell cycle, negative regulation of meiotic cell cycle, and positive regulation of transcription factors. The results showed that the anti-tumor effect of A. annua may be associated with regulating lipid metabolism of tumor cells, decomposing large amounts of lipids, releasing energy, reducing the rate of tumor cell division and accelerating tumor cell apoptosis.

Highly Selective Bioconversion of Ginsenoside Rb1 to Compound K by the Mycelium of Cordyceps sinensis under Optimized Conditions.

Compound K (CK), a highly active and bioavailable derivative obtained from protopanaxadiol ginsenosides, displays a wide variety of pharmacological properties, especially antitumor activity. However, the inadequacy of natural sources limits its application in the pharmaceutical industry. In this study, we firstly discovered that Cordyceps sinensis was a potent biocatalyst for the biotransformation of ginsenoside Rb1 into CK. After a series of investigations on the biotransformation parameters, an optimal composition of the biotransformation culture was found to be lactose, soybean powder and MgSO4 without controlling the pH. Also, an optimum temperature of 30 °C for the biotransformation process was suggested in a range of 25 °C-50 °C. Then, a biotransformation pathway of Rb1→Rd→F2→CK was established using high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS). Our results demonstrated that the molar bioconversion rate of Rb1 to CK was more than 82% and the purity of CK produced by C. sinensis under the optimized conditions was more than 91%. In conclusion, the combination of C. sinensis and the optimized conditions is applicable for the industrial preparation of CK for medicinal purposes.

Dihydromyricetin inhibits migration and invasion of hepatoma cells through regulation of MMP-9 expression.

AIM: To investigate the effects of dihydromyricetin (DHM) on the migration and invasion of human hepatic cancer cells. METHODS: The hepatoma cell lines SK-Hep-1 and MHCC97L were used in this study. The cells were cultured in RPIM-1640 medium supplemented with 10% fetal bovine serum at 37 °C in a humidified 5% CO2 incubator. DHM was dissolved in dimethyl sulfoxide and diluted to various concentrations in medium before applying to cells. MTT assays were performed to measure the viability of the cells after DHM treatment. Wound healing and Boyden transwell assays were used to assess cancer cell motility. The invasive capacity of cancer cells was measured using Matrigel-coated transwell chambers. Matrix metalloproteinase (MMP)-2/9 activity was examined by fluorescence analysis. Western blot was carried out to analyze the expression of MMP-2, MMP-9, p-38, JNK, ERK1/2 and PKC-δ proteins. All data were analyzed by Student's t tests in GraphPad prism 5.0 software and are presented as mean ± SD. RESULTS: DHM was found to strongly inhibit the migration of the hepatoma cell lines SK-Hep-1 (without DHM, 24 h: 120 ± 8 µmol/L vs 100 µmol/L DHM, 24 h: 65 ± 10 µmol/L, P < 0.001) and MHCC97L (without DHM, 24 h: 126 ± 7 µmol/L vs 100 µmol/L DHM, 24 h: 74 ± 6 µmol/L, P < 0.001). The invasive capacity of the cells was reduced by DHM treatment (SK-Hep-1 cells without DHM, 24 h: 67 ± 4 µmol/L vs 100 µmol/L DHM, 24 h: 9 ± 3 µmol/L, P < 0.001; MHCC97L cells without DHM, 24 h: 117 ± 8 µmol/L vs 100 µmol/L DHM, 24 h: 45 ± 2 µmol/L, P < 0.001). MMP2/9 activity was also inhibited by DHM exposure (SK-Hep-1 cells without DHM, 24 h: 600 ± 26 µmol/L vs 100 µmol/L DHM, 24 h: 100 ± 6 µmol/L, P < 0.001; MHCC97L cells without DHM, 24 h: 504 ± 32 µmol/L vs 100 µmol/L DHM 24 h: 156 ± 10 µmol/L, P < 0.001). Western blot analysis showed that DHM decreased the expression level of MMP-9 but had little effect on MMP-2. Further investigation indicated that DHM markedly reduced the phosphorylation levels of p38, ERK1/2 and JNK in a concentration-dependent manner but had no impact on the total protein levels. In addition, PKC-δ protein, a key protein in the regulation of MMP family protein expression, was up-regulated with DHM treatment. CONCLUSION: These findings demonstrate that DHM inhibits the migration and invasion of hepatoma cells and may serve as a potential candidate agent for the prevention of HCC metastasis.

MicroRNA-520c-3p inhibits hepatocellular carcinoma cell proliferation and invasion through induction of cell apoptosis by targeting glypican-3.

AIM: Glypican-3 (GPC3) is a membrane-associated heparan sulfate proteoglycan involved in regulation of cell proliferation, cell survival, cell migration and differentiation process. MicroRNAs (miRNAs) are single-stranded, non-coding functional RNAs that are important in many biological processes. GPC3 and miRNAs have been found to play essential roles in the development and progression of hepatocellular carcinoma (HCC). However, little information about the relationship between GPC3 and miRNAs is available nowadays. Therefore, this study aims to examine the relationship between GPC3 and miRNAs. METHODS: Dual-luciferase reporter assay was used to validate the direct target of GPC3. Fluorescence quantitative PCR and Western blotting were used to examined the gene expression at mRNA and protein levels. Cell apoptosis was evaluated by flow cytometric analysis and Annexin V-FITC staining. Invasion of cells was evaluated by Transwell matrigel assay. RESULTS: The results showed that miR-520c-3p could specifically target GPC3 in HCC cells. GPC3 protein levels decreased with unchanged transcription efficiency after miRNA transfection, and there was negative correlation of miR-520c-3p expression in HCC in relate to GPC3 protein levels. Moreover, miR-520c-3p not only induced HCC cell apoptosis, but also inhibited the growth and invasion of the cells. Interestingly, overexpression of GPC3 could effectively reverse apoptosis induced by miR-520c-3p transfection in HCC. CONCLUSIONS: Taken together, these results supported that miR-520c-3p may decrease GPC3 protein levels to inhibit proliferation of HCC cells. Therefore, GPC3 could be a new target for genetic diagnosis and treatment of HCC.