Targeting Liver Cancer Research Articles

Abstract 1569: Development of a noninvasive and sensitive urine screening test for liver cancer targeting circulation-derived cancer DNA biomarkers

Abstract Hepatocellular carcinoma (HCC), or liver cancer, is an aggressive disease and is regarded as one of the fastest growing cancers in the United States in incidence. The 5-year survival rate of HCC patients drops dramatically from a poor 26%, in early-stage cancer, to a mere 2% in late stages of this disease. The only available early detection biomarker is serum alpha-fetoprotein, which can detect only 40-60% of cases. Our goal is to develop a screening test for early detection of liver cancer, in order to meet this urgent need for new and better biomarkers for HCC. Cancer is a disease of the genome and epigenome, and if we can detect these underlying genetic mutations and epigenetic modifications in the periphery, we can effectively detect cancer early. Hence, we chose a panel of both HCC-associated genetic and DNA methylation biomarkers. We have previously shown that urine contains fragmented cell-free DNA derived from the circulation and that we can detect cancer-related DNA, both mutated and methylated DNA, in the urine of such patients. We have also demonstrated that the concentration of tumor-derived DNA in plasma and in urine is similar in patients with tumors. In order to demonstrate the feasibility of detecting HCC DNA biomarkers in the circulation derived urine DNA, we have developed short amplicon (∼50 bp) PCR-based assays targeting HCC-specific mutations in TP53 and CTNNB1 genes and HCC-specific methylation in GSTP1, RASSF1A and CDKN2A genes. Urine DNA isolated from samples of hepatitis, cirrhosis, and HCC patients were tested in each of the assays and analyzed for individual marker and panel performance. The performance of the markers and our approach towards the development of a targeted next generation urine DNA based HCC screening test is discussed. Citation Format: Surbhi Jain, Sitong Chen, Selena Y. Lin, Adam Clemens, Hei-won L. Hann, Ting-Tsung Chang, Chi-Tan Hu, Shun-Hua Chen, Wei Song, Ying-Hsiu Su. Development of a noninvasive and sensitive urine screening test for liver cancer targeting circulation-derived cancer DNA biomarkers. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1569. doi:10.1158/1538-7445.AM2015-1569

Pravastatin chitosan nanogels-loaded erythrocytes as a new delivery strategy for targeting liver cancer

Chitosan nanogels (CNG) are developed as one of the most promising carriers for cancer targeting. However, these carriers are rapidly eliminated from circulation by reticuloendothelial system (RES), which limits their application. Therefore, erythrocytes (ER) loaded CNG as multifunctional carrier may overcome the massive elimination of nanocarriers by RES. In this study, erythrocytes loaded pravastatin–chitosan nanogels (PR–CNG–ER) were utilized as a novel drug carrier to target liver cancer. Thus, PR–CNG formula was developed in nanosize, with good entrapment efficiency, drug loading and sustained release over 48h. Then, PR–CNG loaded into ER were prepared by hypotonic preswelling technique. The resulting PR–CNG–ER showed 36.85% of entrapment efficiency, 66.82% of cell recovery and release consistent to that of hemoglobin over 48h. Moreover, PR–CNG–ER exhibited negative zeta potential, increasing of hemolysis percent, marked phosphatidylserine exposure and stomatocytes shape compared to control unloaded erythrocytes. PR–CNG–ER reduced cells viability of HepG2 cells line by 28% compared to unloaded erythrocytes (UER). These results concluded that PR–CNG–ER are promising drug carriers to target liver cancer.

Synergistic inhibition of autophagy and neddylation pathways as a novel therapeutic approach for targeting liver cancer.

Liver cancer is the second-most frequent cause of cancer death in the world and is highly treatment resistant. We reported previously that inhibition of neddylation pathway with specific NAE inhibitor MLN4924, suppressed the malignant phenotypes of liver cancer. However, during the process, MLN4924 induces pro-survival autophagy as a mechanism of drug resistance. Here, we report that blockage of autophagy with clinically-available autophagy inhibitors (e.g. chloroquine) significantly enhanced the efficacy of MLN4924 on liver cancer cells by triggering apoptosis. Mechanistically, chloroquine enhanced MLN4924-induced up-regulation of pro-apoptotic proteins (e.g. NOXA) and down-regulation of anti-apoptotic proteins. Importantly, the down-regulation of NOXA expression via siRNA silencing substantially attenuated apoptosis of liver cancer cells. Further mechanistic studies revealed that blockage of autophagy augmented MLN4924-induced DNA damage and reactive oxygen species (ROS) generation. The elimination of DNA damage or blockage of ROS production significantly reduced the expression of NOXA, and thereby attenuated apoptosis and reduced growth inhibition of liver cancer cells. Moreover, blockage of autophagy enhanced the efficacy of MLN4924 in an orthotopic model of human liver cancer, with induction of NOXA and apoptosis in tumor tissues. These findings provide important preclinical evidence for clinical investigation of synergistic inhibition of neddylation and autophagy in liver cancer.

Induction of human UDP-Glucuronosyltransferase 2B7 gene expression by cytotoxic anticancer drugs in liver cancer HepG2 cells.

We recently reported induction of UGT2B7 by its substrate epirubicin, a cytotoxic anthracycline anticancer drug, via activation of p53 and subsequent recruitment of p53 to the UGT2B7 promoter in hepatocellular carcinoma HepG2 cells. Using the same HepG2 model cell line, the present study assessed the possibility of a similar induction of UGT2B7 by several other cytotoxic drugs. We first demonstrated by reverse transcriptase quantitative real-time polymerase chain reaction that, as observed with epirubicin, nine cytotoxic drugs including three anthracyclines (doxorubicin, daunorubicin, and idarubicin) and six nonanthracyclines (mitomycin C, 5-fluorouracil, camptothecin, 7-ethyl-10-hydroxycamptothecin, topotecan, and etoposide) significantly increased UGT2B7 mRNA levels. To investigate a potential involvement of p53 in this induction, we conducted further experiments with four of the nine drugs (doxorubicin, daunorubicin, idarubicin, and mitomycin C). The cytotoxic drugs studied increased p53 and UGT2B7 protein levels. Knockdown of p53 expression by small interfering RNA reduced cytotoxic drug-induced UGT2B7 expression. Luciferase reporter assays showed activation of the UGT2B7 promoter by cytotoxic drugs via a previously reported p53 site. Finally, chromatin immunoprecipitation assays demonstrated p53 recruitment to the UGT2B7 p53 site upon exposure to mitomycin C, the most potent UGT2B7 inducer among the nine tested drugs. Taken together, these results provide further evidence supporting UGT2B7 as a p53 target gene. The cytotoxic drug-induced UGT2B7 activity in target liver cancer cells or possibly in normal liver cells may affect the therapeutic efficacy of co-administered cytotoxic drugs (e.g., epirubicin) and noncytotoxic drugs (e.g., morphine), which are UGT2B7 substrates.

The clinical application of near-infrared light imaging mediated by indocyanine green in liver cancer surgery

In recent years, the emerging optical imaging technologies have shown their unique value in diagnosis of disease and surgical navigation. This article focus on describing the optical imaging technology mediated by the only one near-infrared light imaging molecule--indocyanine green which has already been approved by FDA to use in clinical and inmates a characteristics of passive targeting liver cancer tissue. This article reviews the application of the technology in liver cancer surgery and demonstrates the value, deficiency and possible improvements of this technology.

Stimuli-responsive PEGylated prodrugs for targeted doxorubicin delivery.

In recent years, stimuli-sensitive prodrugs have been extensively studied for the rapid "burst" release of antitumor drugs to enhance chemotherapeutic efficiency. In this study, a novel stimuli-sensitive prodrug containing galactosamine as a targeting moiety, poly(ethylene glycol)-doxorubicin (PEG-DOX) conjugate, was developed for targeting HepG2 human liver cancer cells. To obtain the PEG-DOX conjugate, both galactosamine-decorated poly(ethylene glycol) aldehyde (Gal-PEG-CHO) and methoxy poly(ethylene glycol) aldehyde (mPEG-CHO) were firstly synthesized and functionalized with dithiodipropionate dihydrazide (TPH) through direct reductive amination via Schiff's base formation, and then DOX molecules were chemically conjugated to the hydrazide end groups of TPH-functionalized Gal-/m-PEG chains via pH-sensitive hydrazone linkages. The chemical structures of TPH-functionalized PEG and PEG-DOX prodrug were confirmed by (1)H NMR analysis. The PEG-DOX conjugate could self-assemble into spherical nanomicelles with a mean diameter of 140 nm, as indicated by transmission electron microscopy and dynamic light scattering. The drug loading content and loading efficiency in the prodrug nanomicelles were as high as 20 wt.% and 75 wt.%, respectively. In vitro drug release studies showed that DOX was released rapidly from the prodrug nanomicelles at the intracellular levels of pH and reducing agent. Cellular uptake and MTT experiments demonstrated that the galactosamine-decorated prodrug nanomicelles were more efficiently internalized into HepG2 cells via a receptor-mediated endocytosis process and exhibited a higher toxicity, compared with pristine prodrug nanomicelles. These results suggest that the novel Gal-PEG-DOX prodrug nanomicelles have tremendous potential for targeted liver cancer therapy.

Racial/ethnic disparities in liver cancer-attributable hepatitis infections in the U.S. population.

259 Background: According to the American Cancer Association, hepatitisB and Cinfections may raise liver cancer risk. We identify population-specific cancer-attributable hepatitis prevalence to discern disparities across race/ethnicity for liver cancer risk. Methods: Data from 1999-2012 National Health and Nutrition Examination Survey (NHANES) were used to determine prevalence, unadjusted odds ratios (OR) with 95% confidence intervals (95%CI) for non-Hispanic-Whites (NHW), non-Hispanic-Blacks (NHB), Mexican-Americans, other Hispanics, and other race of hepatitis, taking complex sampling design into account. Results: Among those sampled, the overall highest prevalence of hepatitis infection was 66.4% in NHW for hepatitis-C and the lowest prevalence was 1.4% in Mexican-Americans for hepatitis-B surface-antigen. NHB were more likely than NHW to have hepatitis-B infections of surface-antibody (OR=1.9; 95%CI=1.8-2.0), core-antibody (3.3; 3.1-3.6), surface-antigen (5.3; 3.6-7.7), and hepatitis-C infections (1.7; 1.5-2.0). Compared to NHW, other Hispanics had a higher burden of hepatitis-B infection of surface-antibody (1.5; 1.4-1.7) and core-antibody (2.5; 2.1-2.8); other race had the same higher infection pattern as other Hispanics in surface-antibody (2.4; 2.2-2.6) and core-antibody (6.7; 6.0-7.5). Surface-antibody was more prevalent (1.5; 1.4-1.6) while core-antibody was less prevalent (0.9; 0.8-0.9) among Mexican-Americans compared with NHW. Conclusions: The prevalence of hepatitis B and C infections varies across different race/ethnicities with NHB having the greatest burden compared with NHW. Population-based databases should collect more information regarding infection burden in at-risk groups in order to understand more about these disparities. Appreciating such gaps across demographics could assist in future efforts to decrease the burden of hepatitis and to provide a base for targeted liver cancer prevention programs via screening, vaccination, health promotion, and literacy.

Multifunctional lactobionic acid-modified dendrimers for targeted drug delivery to liver cancer cells: investigating the role played by PEG spacer.

We report the development of a lactobionic acid (LA)-modified multifunctional dendrimer-based carrier system for targeted therapy of liver cancer cells overexpressing asialoglycoprotein receptors. In this study, generation 5 (G5) poly(amidoamine) (PAMAM) dendrimers were sequentially modified with fluorescein isothiocyanate (FI) and LA (or polyethylene glycol (PEG)-linked LA, PEG-LA), followed by acetylation of the remaining dendrimer terminal amines. The synthesized G5.NHAc-FI-LA or G5.NHAc-FI-PEG-LA conjugates (NHAc denotes acetamide groups) were used to encapsulate a model anticancer drug doxorubicin (DOX). We show that both conjugates are able to encapsulate approximately 5.0 DOX molecules within each dendrimer and the formed dendrimer/DOX complexes are stable under different pH conditions and different aqueous media. The G5.NHAc-FI-PEG-LA conjugate appears to have a better cytocompatibility, enables a slightly faster DOX release rate, and displays better liver cancer cell targeting ability than the G5.NHAc-FI-LA conjugate without PEG under similar experimental conditions. Importantly, the developed G5.NHAc-FI-PEG-LA/DOX complexes are able to specifically inhibit the growth of the target cells with a better efficiency than the G5.NHAc-FI-LA/DOX complexes at a relatively high DOX concentration. Our results suggest a key role played by the PEG spacer that affords the dendrimer platform with enhanced targeting and therapeutic efficacy of cancer cells. The developed LA-modified multifunctional dendrimer conjugate with a PEG spacer may be used as a delivery system for targeted liver cancer therapy and offers new opportunities in the design of multifunctional drug carriers for targeted cancer therapy applications.

Preparation of camptothecin-loaded targeting nanoparticles and their antitumor effects on hepatocellular carcinoma cell line H22

Camptothecin (CPT) is an effective anticancer agent against various cancers but the clinical application is limited because of its poor water solubility, low bioavailability and severe toxic side effects. The aim of the present study was to evaluate the feasibility of using targeted NPs as a high-performance CPT delivery system that targets liver cancer cells through intravenous (i.v.) administration route. CPT was incorporated into biotin-F127-PLA or F127-PLA polymeric nanoparticles (NPs) by a dialysis method. The preparation of the targeting NPs was performed by conjugating biotin-F127-PLA NPs with anti-3A5 antibody. The antitumor effect of the CPT-loaded nanoparticles against H22 cells in vitro was determined using an MTT assay. Tissue distribution and tumor inhibition in vivo were also evaluated. Survivin mRNA expression was assessed by real-time polymerase chain reaction. Results showed that the targeted CPT NPs exhibited regular spherical shapes with a mean diameter of approximately 180 nm. In vitro release of the targeted CPT NPs exhibited an initial burst (40%) within 12 h, followed by a slow release. Cytotoxicity test against H22 cells indicated that the targeted CPT NPs exerted significant antitumor effects. Compared with free CPT and non-targeted CPT NPs, the targeted CPT NPs showed superior inhibition ratio against tumor in vivo, which may be associated with reduced survivin mRNA expression. The results suggested that the new targeted CPT NPs may be a promising injectable delivery system for cancer therapy.

Diagnosis applications of new hepatoma carcinoma cell aptamers in vitro

In this study, we used the “cell-SELEX” strategy to generate four new aptamers with high binding affinity for Huh7.5.1. and HepG2 cells, as indicated by Kd values in the low nanomolar range, as well as high specificity compared to L-02 normal human liver cells. Moreover, the aptamers could distinguish human liver tumour tissues from normal tissues. Incorporation of the aptamer into probes permitted activation by target liver cancer cells to yield enhanced fluorescence and the detection of 103 orders of target liver cancer cells. The newly generated aptamers have great potential in early cancer diagnosis.

Liver cancer cells: targeting and prolonged-release drug carriers consisting of mesoporous silica nanoparticles and alginate microspheres.

A new microsphere consisting of inorganic mesoporous silica nanoparticles (MSNs) and organic alginate (denoted as MSN@Alg) was successfully synthesized by air-dynamic atomization and applied to the intracellular drug delivery systems (DDS) of liver cancer cells with sustained release and specific targeting properties. MSN@Alg microspheres have the advantages of MSN and alginate, where MSN provides a large surface area for high drug loading and alginate provides excellent biocompatibility and COOH functionality for specific targeting. Rhodamine 6G was used as a model drug, and the sustained release behavior of the rhodamine 6G-loaded MSN@Alg microspheres can be prolonged up to 20 days. For targeting therapy, the anticancer drug doxorubicin was loaded into MSN@Alg microspheres, and the (lysine)4-tyrosine-arginine-glycine-aspartic acid (K4YRGD) peptide was functionalized onto the surface of MSN@Alg for targeting liver cancer cells, hepatocellular carcinoma (HepG2). The results of the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay and confocal laser scanning microscopy indicate that the MSN@Alg microspheres were successfully uptaken by HepG2 without apparent cytotoxicity. In addition, the intracellular drug delivery efficiency was greatly enhanced (ie, 3.5-fold) for the arginine-glycine-aspartic acid (RGD)-labeled, doxorubicin-loaded MSN@Alg drug delivery system compared with the non-RGD case. The synthesized MSN@Alg microspheres show great potential as drug vehicles with high biocompatibility, sustained release, and targeting features for future intracellular DDS.

Abstract B97: AS1411-Doxorubicin adduct for targeted liver cancer therapy.

Abstract Background: Aptamer-drug conjugates are promising to be applied to targeted cancer therapy, in which anticancer drugs were selectively delivered into cognate cancerous cells, but not to healthy tissues, in order to reduce side effects and enhance therapeutic efficacy. Methodology/Principal Findings: In this study, the technology of drug-DNA adduct (DDA) was utilized to prepare drug-aptamer conjugates. Particularly, an anticancer drug, Doxorubicin (Dox), was conjugated with AS1411, a DNA aptamer which specifically recognizes human hepatocellular carcinoma Huh7 cells. The technology enabled a simple and high-payload conjugation of drugs onto aptamers. The resultant AS1411-Dox adduct maintained selectively recognition ability to target liver cancer cells with strong binding affinity (demonstrated by flow cytometry), was able to selectively deliver the conjugated Dox into target cancer cells (observed with confocal microscopy), and exhibited selective and potent cytotoxicity in target liver cancer cells (evaluated by MTS cell viability assay). More importantly, as the first of its kind, AS1411-Dox adduct demonstrated selectively antitumor therapeutic efficacy of this drug-aptamer conjugation technology in a mouse model. The ability of drug-aptamer adducts to deliver anticancer drugs into target cancer cells, while inhibiting nonspecific drug uptake in nontarget cells, enables this technology promising to be implemented in targeted cancer therapy. Conclusions/Significance: Given the demonstrated ability of targeted liver cancer therapy combined with the aforementioned advantageous features as a simple preparation, the high drug payload capacity, and the wide applicability, this novel drug-DNA adduct technology is anticipated for wide implications for targeted cancer therapy. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):B97. Citation Format: Thu Le Trinh, Guizhi Zhu, Xilin Xiao, Xiaokui Zhang, Kwame Sefah, Weihong Tan, Chen Liu. AS1411-Doxorubicin adduct for targeted liver cancer therapy. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr B97.

Paclitaxel-loaded nanoparticles decorated with anti-CD133 antibody for targeting liver cancer stem cells

Paclitaxel-loaded nanoparticles decorated with anti-CD133 antibody for targeting liver cancer stem cells

A pH‐responsive drug release system based on doxorubicin conjugated amphiphilic polymer coated quantum dots for tumor cell targeting and tracking

AbstractBACKGROUNDA tumor‐targeted and pH‐responsive drug release system based on poly(ethylene glycol) (PEG) and dodecylamine (DDA)‐modified poly itaconic acid (PIA) with doxorubicin (DOX) and quantum dots (QDs) (PIA‐PEG‐DDA‐DOX@QDs) has been constructed successfully. These nanocomposites with outstanding capabilities were further designed to target liver cancer cells with vascular endothelial growth factor receptor (VEGFR) overexpressed.RESULTSThe nanoparticles constructed were spherical and monodispersed with very small diameter. The cumulative release rates of DOX conjugates were studied at pH 5.5 and pH 7.4, and the release behavior at pH 5.5 fit the first‐order kinetics model, Q=46.5991 –45.2394e‐0.24355t for PIA‐PEG‐DDA‐DOX and Q=46.7094–38.5588e‐0.20123t for PIA‐PEG‐DDA‐DOX@QDs. The nanoparticles show low cytotoxicity at pH 7.4 and high cytotoxicity at pH 5.5 in cell viability studies. In the intracellular localization observation, the targeting molecule anti‐VEGF conjugated NPs exhibited efficient receptor‐mediated endocytosis in anti‐VEGF receptor‐overexpressing cancer cells, compared with nontargeted nanoparticles.CONCLUSIONSThe targeting imaging and pH‐responsive drug release system could target the tumor cells and fluoresce quantum dots sensitively due to the conjugation of VEGF, and enabled pH‐controlled‐activation of the DOX. The water soluble conjugates have the potential of becoming a promising diagnostic tool in clinical application for cancer detection and treatment. © 2013 Society of Chemical Industry

Lactose substituted zinc phthalocyanine: A near infrared fluorescence imaging probe for liver cancer targeting

A near infrared fluorescence probe, lactose substituted zinc phthalocyanine, [2,9(10),16(17),23(24)-tetrakis((1-(β-d-lactose-2-yl)-1H-1,2,3-triazol-4-yl)methoxyl)phthalocyaninato] zinc(II), was synthesized via click reaction. Structural characterization and optical experiment demonstrated its excellent biocompatibility and fluorescence imaging ability. Near infrared fluorescence imaging in vivo for liver cancer, lung cancer and melanoma cancer with tumor bearing nude mice as models demonstrated that lactose substituted zinc phthalocyanine has specifically targeting ability to liver cancer while no targeting to lung cancer or melanoma, which implied its potential in liver cancer diagnosis as a near infrared optical probe.

A novel microfluidic co-culture system for investigation of bacterial cancer targeting

Although bacterial cancer targeting in animal models has been previously demonstrated and suggested as a possible therapeutic tool, a thorough understanding of the mechanisms responsible for cancer specificity would be required prior to clinical applications. To visualize bacterial preference for cancer cells over normal cells and to elucidate the cancer-targeting mechanism, a simple microfluidic platform has been developed for in vitro studies. This platform allows simultaneous cultures of multiple cell types in independent culture environments in isolated chambers, and creates a stable chemical gradient across a collagen-filled passage between each of these cell culture chambers and the central channel. The established chemical gradient induces chemotactic preferential migration of bacteria toward a particular cell type for quantitative analysis. As a demonstration, we tested differential bacterial behavior on a two-chamber device where we quantified bacterial preference based on the difference in fluorescence intensities of green fluorescence protein (GFP)-expressing bacteria at two exits of the collagen-filled passages. Analysis of the chemotactic behavior of Salmonella typhimurium toward normal versus cancer hepatocytes using the developed platform revealed an apparent preference for cancer hepatocytes. We also demonstrate that alpha-fetoprotein (AFP) is one of the key chemo-attractants for S. typhimurium in targeting liver cancer.

Artifact-reduced simultaneous MRI of multiple rats with liver cancer using PROPELLER

To explore simultaneous magnetic resonance imaging (MRI) for multiple hepatoma-bearing rats in a single session suppressing motion- and flow-related artifacts to conduct preclinical cancer research efficiently. Our institutional Animal Experimental Committee approved this study. We acquired PROPELLER (periodically rotated overlapping parallel lines with enhanced reconstruction) T2 - and diffusion-weighted images of the liver in one healthy and 11 N1-S1 hepatoma-bearing rats in three sessions using a 3-T clinical scanner and dedicated multiarray coil. We compared tumor volumes on MR images and those on specimens, evaluated apparent diffusion coefficients (ADC) of the tumor, and compared them to previously reported values. Each MRI session took 39-50 minutes from anesthesia induction to the end of scans for four rats (10-13 minutes per rat). PROPELLER provided artifact-reduced T2 - and diffusion-weighted images of the rat livers. Tumor volumes on MR images ranged from 0.04-1.81 cm(3) and were highly correlated with those on specimens. The ADC was 1.57 ± 0.37 × 10(-3) mm(2) /s (average ± SD), comparable to previously reported values. PROPELLER allowed simultaneous acquisition of artifact-reduced T2 - and diffusion-weighted images of multiple hepatoma-bearing rats. This technique can promote high-throughput preclinical MR research for liver cancer.

Label-free quantitative proteomics of CD133-positive liver cancer stem cells

BackgroundCD133-positive liver cancer stem cells, which are characterized by their resistance to conventional chemotherapy and their tumor initiation ability at limited dilutions, have been recognized as a critical target in liver cancer therapeutics. In the current work, we developed a label-free quantitative method to investigate the proteome of CD133-positive liver cancer stem cells for the purpose of identifying unique biomarkers that can be utilized for targeting liver cancer stem cells. Label-free quantitation was performed in combination with ID-based Elution time Alignment by Linear regression Quantitation (IDEAL-Q) and MaxQuant.ResultsInitially, IDEAL-Q analysis revealed that 151 proteins were differentially expressed in the CD133-positive hepatoma cells when compared with CD133-negative cells. We then analyzed these 151 differentially expressed proteins by MaxQuant software and identified 10 significantly up-regulated proteins. The results were further validated by RT-PCR, western blot, flow cytometry or immunofluorescent staining which revealed that prominin-1, annexin A1, annexin A3, transgelin, creatine kinase B, vimentin, and EpCAM were indeed highly expressed in the CD133-positive hepatoma cells.ConclusionsThese findings confirmed that mass spectrometry-based label-free quantitative proteomics can be used to gain insights into liver cancer stem cells.

Recent advances of miRNA involvement in hepatocellular carcinoma and cholangiocarcinoma

MicroRNAs (miRNAs), which are a class of highly evolutionarily conserved non-coding RNAs, modulate gene expression and are regulated by specific genes. Several studies have shown that the expression of miRNAs is deregulated in Hepatitis C virus (HCV) & Hepatitis B virus (HBV) infection, liver cancer progression, tumor invasion and metastasis. There are a number of high-quality review articles relative to the general role of miRNA alterations in carcinogenesis and specific reviews dealing with the miRNA changes in hepatocellular carcinoma (HCC) and cholangio-carcinoma (CCA). Since primary liver cancer is predominantly comprised of HCC and intrahepatic cholangiocarcinoma (ICC), in the present review we specifically focus on recent advances of miRNAs related to tumorigenesis, invasion and metastasis of primary liver cancer, with special emphasis on their relationships to their target genes. HCV & HBV are major causes of liver disease, including acute and chronic hepatitis, liver cirrhosis, and HCC, while HCV infection is a risk factor for ICC. We also discuss the mi-RNA alterations involved in HCV & HBV infection. We briefly describe advances in molecular signaling of miRNAs in liver cancers and present insights into new therapeutic clues that target liver cancer.

Optimizing conditions for encapsulation of QDs by varying PEG chain density of amphiphilic centipede-like copolymer coating and exploration of QDs probes for tumor cell targeting and tracking

Quantum dots (QDs) are potential probes for cell labeling and tracking. But the design of safe and tumor cell-sensitive probes together with high luminescent efficiency is still challenging. In this study, a novel class of biodegradable amphiphilic centipede-like copolymers was synthesized for QDs surface modification, which transformed hydrophobic QDs into hydrophilic ones. A systematic study was carried out to find how poly(ethyleneglycol) (PEG) chain density of the polymer coating influenced the properties of QDs nanocomposites, including particle size, optical properties and morphology. Results indicated that when the PEG grafting level was 40%, QDs nanocomposites showed the smallest size (105 nm), and simultaneously exhibited superior fluorescent properties among all the nanocomposites. These shell–core spherical structural nanoparticles were well dispersed in water, and did not form large aggregates. Apparently, a polymer with a PEG chain density of 40% was the optimum surface coating for QDs. These nanocomposites with outstanding capabilities can be further designed to target liver cancer cells with vascular endothelial growth factor receptor (VEGFR) overexpressed. We have evaluated QD-anti-vascular endothelial growth factor (VEGF) bioconjugates and QDs nanocomposites without anti-VEGF in live cell imaging experiments and results showed that the targeting capability to HepG2 cells of QD-anti-VEGF bioconjugates was more significant than that of non-anti-VEGF QDs. Our results provide a basis for the optimization of the polymer coating of QDs and are important for the design and development of nanoprobes for optical detection and bio-imaging of tumor cells.