Liver Cancer Cell Line SMMC-7721 Research Articles

Evaluating Membrane Electrical Properties of SMMC7721 Cells with TiO₂ NPs Applications to Cytotoxicity by Dielectric Spectroscopy.

Titanium dioxide nanoparticles (TiO₂ NPs) represent one of the most frequently applied nanomaterials in numerous areas of daily life. Recent studies show that TiO₂ exposure increases the occupational risk of liver injury and inflammation, and even liver cancer to the workers of factories handling these NPs. However, the potential risks and biophysical effects of TiO₂ on hepatic cells need extensive evaluation. To this end, we explored the electrophysiological changes in the human liver cancer cell line SMMC7721 following exposure to TiO₂ NPs. TiO₂ NPs decreased the first (Δε1) and second dielectric relaxation intensity (Δε₂) of the SMMC7721 cells by 6.62% and 0.86% respectively, and significantly increased the first characteristic frequency (fc1, 4.82%) and the first Cole-Cole parameter (β1, 1.24%). The double spherical-shell model showed that TiO₂ NPs significantly lowered the permittivity of unit-membrane and capacitance, as well as the conductivity of extracellular fluid, cytoplasm, and nuclear contents compared to the untreated control. Conclusively, this study revealed that TiO₂ NPs induce cytotoxic effects by disrupting the permeability and electrical conductivity of unit membranes. Further, we report that dielectric spectrum combined with model parameter analysis can evaluate the bioelectrical effects of TiO₂ NPs on human liver cancer cells.

6,12-Diphenyl-3, 9-diazatetraasterane-1, 5, 7, 11-tetracarboxylate Inhibits Proliferation, Migration and Promotes Apoptosis in Ovarian Cancer Cells.

6,12-Diphenyl-3,9-diazatetraasterane-1, 5, 7, 11-tetracarboxylate (DDTC) has been synthesized by the photodimerization of 4-phenyl-1,4-dihydropyridine-3,5-dicarboxylate. The potential of theercvantitumor activity and mechanism were investigated in vitro using MTT assay in human lung cancer cell line A549, ovarian cancer cell lines SKOV3 and A2780, breast cancer cell line MCF-7, gastric cancer cell line BGC-823, colon cancer cell line HT29, prostate cancer cell line DU145, and liver cancer cell line SMMC7721. The results show that DDTC can inhibit the growth of ovarian cancer SKOV3 and A2780 cells. The best IC50 value is approximately 5.29 ± 0.38 and 4.29 ± 0.39 μM, respectively. DDTC induced the cell cycle arrest in the G2 phase by flow cytometric analysis. The migration and invasion of ovarian cancer SKOV3 and A2780 cells were inhibited by DDTC. DDTC could increase the expression protein level of E-cadherin in A2780 cells and ascend the expression protein and mRNA levels of E-cadherin in SKOV3 cells. DDTC could also decrease the protein and mRNA expression of EMT (epithelial-to-mesenchymal transition) markers of N-cadherin and Vimentin. mRNA and protein expression level of checkpoint kinase 1 (Chk1) were significantly increased and expressions of cyclin-dependent kinase (CDK1) and cell division cycle 25a (Cdc25a) were decreased in the SKOV3 and A2780 cell lines. Moreover, DDTC induced apoptosis by the cleavage and activation of caspase 3 and caspase 9.

Silence of cytoskeleton-associated protein 2 represses cell proliferation and migration and promotes apoptosis in liver cancer cell lines.

To investigate the roles of cytoskeleton-associated protein 2 (CKAP2) in proliferation, apoptosis, and migration in liver cancer cells and the potential mechanisms. Human normal hepatocyte L02 and liver cancer cell lines HepG2, Huh7, and SMMC-7721 were cultured. The CKAP2 expression was detected by real-time PCR and Western blotting. HepG2 cells were randomly divided into a control group, a negative control (NC) group, and a CKAP2 silencing (siCKAP2) group. CCK-8 and BrdU assays were used to evaluate cell viability and proliferation, respectively. Transwell assay was employed to determine cell migration and invasion. The protein levels of cleaved-caspase 3, Bax, E-cadherin, N-cadherin, Vimentin, phosphorylated Janus kinase 2 (p-JAK2), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) were determined by Western blotting. Compared with normal hepatocyte L02, CKAP2 was highly expressed in liver cancer cell lines HepG2, Huh7, and SMMC-7721 (all P<0.05). Compared with the NC group, cell viability and proliferation rate of the siCKAP2 group were decreased (both P<0.05). The apoptotic rate, protein expression of cleaved-caspase 3 and Bax in the siCKAP2 group were significantly higher than those in the NC group (all P<0.05). Compared with the NC group, cell migration and invasion rates of the siCKAP2 group were significantly attenuated (both P<0.05). Compared with the NC group, E-cadherin protein expression in siCKAP2 group was increased, while protein expression levels of Vimentin, N-cadherin, p-JAK2, and p-STAT3 were decreased (all P<0.05). CKAP2 gene silence inhibits proliferation, migration, and invasion, and promotes apoptosis in liver cancer cells, while JAK2/STAT3 signaling pathway may be involved in these processes.

EGFRvIII-specific CAR-T cells produced by piggyBac transposon exhibit efficient growth suppression against hepatocellular carcinoma.

Adoptive cellular immunotherapy employing chimeric antigen receptors-modified T (CAR-T) cells has demonstrated promising antitumor effects in hematologic cancers. However, CAR-T therapy confront many challenges in solid tumors like immunosuppressive microenvironment, molecular heterogeneity, etc. The cancer genome atlas (TCGA) of hepatocellular carcinoma (HCC) revealed many genetic characteristic and molecular tumorigenesis. EGFRvIII is a tumor specific antigen widely expressed in a variety of cancers including HCC and an ideal therapeutic target for cancer therapy. The liver cancer cell line SMMC7721 express high level EGFRvIII and widely applied in HCC investigations. Herein, we developed EGFRvIII CAR-T cells by piggyBac transposon system, and detected its specific killing effect against SMMC7721 cells in vitro and in vivo. Results indicated that transduction efficiency of CAR reached 53.1%. Expression of CAR protein was verified by immunoblotting as a band of approximate 57KD. The killing effect of CAR-T cells against SMMC7721 was positively correlated with E/T ratio (E:T=5:1, 10:1, 20:1, 40:1), and exceeded 50% at 20:1 ratio. Significant increase in IFN-γ and TNF-α secretion were detected in the co-culture supernatant of CAR-T cells and SMMC7721, comparable to the level of exogenous EGFRvIII-expressing U87 cells. The killing activity and cytokine secretion were both dependent on the expression level of EGFRvIII in target cells. In HCC xenograft models, CAR-T cells could effectively suppress the growth of SMMC7721. In conclusion, EGFRvIII CAR-T cells demonstrated specific antitumor effect against SMMC7721 in vitro and in vivo, providing basis for immunotherapy of HCC in future clinical use.

LncRNA SNHG1 promotes liver cancer development through inhibiting p53 expression via binding to DNMT1.

This study aims to investigate whether long non-coding RNA (lncRNA) SNHG1 could regulate proliferative and invasive abilities of liver cancer (LC) cells via p53 and DNMT1, so as to regulate the occurrence and progression of LC. SNHG1 expression in LC tissues and paracancerous tissues was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Correlation between SNHG1 expression and tumor stage of LC patients was analyzed. The regulatory effects of SNHG1 and p53 on proliferative, invasive capacities and cell cycle were accessed by CCK-8 (cell counting kit-8), transwell assay and flow cytometry, respectively. The binding condition between SNHG1 and DNMT1 was determined by RNA binding protein immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP). Western blot was conducted to determine whether SNHG1 could regulate p53 in LC cells. Finally, rescue experiments were carried out to evaluate whether SNHG1 regulates proliferative and invasive abilities of LC cells through p53. SNHG1 expression was higher in LC tissues than that of paracancerous tissues. LC patients with stage III-IV presented higher expression level of SNHG1 than those with stage I-II. Similarly, SNHG1 was highly expressed in LC cells than that of normal liver cells. LC cell lines SMMC-7721 and SK-HEP-1 were selected for this study. SNHG1 knockdown inhibited the proliferative and invasive abilities, and arrested the cell cycle in the G0/G1 phase of SMMC-7721 and SK-HEP-1 cells. RIP and ChIP results demonstrated that SNHG1 could bind to DNMT1 and inhibit p53 expression. Overexpression of p53 partially reversed the inhibitory effects of SNHG1 on proliferative and invasive abilities of LC cells. High expression of SNHG1 could promote proliferative and invasive abilities of LC cells through targeting inhibition of p53 expression by binding to DNMT1.

MRC-5 Cancer-associated Fibroblasts Influence Production of Cancer Stem Cell Markers and Inflammation-associated Cell Surface Molecules, in Liver Cancer Cell Lines.

Background: Current opinion suggests that expansion of cancer stem cells (CSCs) and activation of pro-tumoral inflammation cascade correlate with cancer progression.Materials and methods: We explored the possible contributions of MRC-5 cancer-associated fibroblasts to the expression profiles of CSC markers and inflammation-associated cell surface molecules. The liver cancer cell lines Bel-7402, SMMC-7721, MHCC-LM3, and HepG2 cultured in conditioned medium (CM) from MRC-5 served as test groups, whereas the liver cancer cell lines cultured in normal medium served as control groups.Results: Flow cytometry revealed that the proportions of CD90+ cells were significantly higher in MHCC-LM3-(MRC-5)-CM and HepG2-(MRC-5)-CM cells, and moderately higher in Bel-7402-(MRC-5)-CM and SMMC-7721-(MRC-5)-CM cells, than in controls. The CD90+/CD45- proportions were elevated in Bel-7402-(MRC-5)-CM and MHCC-LM3-(MRC-5)-CM cells, but reduced in HepG2-(MRC-5)-CM and SMMC-7721-(MRC-5)-CM cells, as compared to controls. Western blotting indicated that Nanog was downregulated in MHCC-LM3-(MRC-5)-CM and HepG2-(MRC-5)-CM cells, compared to controls; that POU5F1 (OCT4/3) was downregulated in MHCC-LM3-(MRC-5)-CM, but upregulated in Bel-7402-(MRC-5)-CM and HepG2-(MRC-5)-CM cells, compared to controls, and that CK19 was upregulated in Bel-7402-(MRC-5)-CM and MHCC-LM3-(MRC-5)-CM cells, compared to controls. Proportions of cells expressing Toll-like receptor-1+ (TLR1) and TLR4 were significantly higher in MHCC-LM3-(MRC-5)-CM cells, and moderately higher in HepG2-(MRC-5)-CM cells, than controls. However, the TLR1+ and TLR4+ proportions were lower in Bel-7402-(MRC-5)-CM and SMMC-7721-(MRC-5)-CM cells than controls. Proportions of CD25+ cells were reduced in HepG2-(MRC-5)-CM and SMMC-7721-(MRC-5)-CM cells, but elevated in MHCC-LM3-(MRC-5)-CM and Bel-7402-(MRC-5)-CM cells, compared to controls. Proportion of CD61+ cells was higher in liver cancer cells cultured in MRC-5-CM than in controls. Proportion of CD14+ cells was lower in HCC cells cultured in MRC-5-CM than in controls.Conclusion: MRC-5 extensively affected the production of CSC markers and inflammation-associated cell surface molecules. Tumor-targeting molecular therapies should consider these findings.

The influence of two different cultivations on human liver cancer cell line SMMC-7721 cells

Objective To observe and compare thechange of human liver cancer cell line SMMC-7721 cells’ related characteristics using adherent cultivation and suspension cultivation. Methods (1) To collect human liver cancer cell line SMMC-7721 cells cultured with ordinary medium (adherent cultivation) and with serum-free medium (suspension cultivation). (2) To observe morphologic patterns of SMMC-7721 tumor cells treated with two kinds of cultivationunder inverted microscope. (3) To measure expressions of tumor stem cell surface markers CD90, CD133 and CD24 in SMMC-7721 tumor cells treated with two kinds of cultivation by the flow cytometry. (4) To detect mRNA expressions of the tumor stem cell surface markers CD90, CD133 and CD24 in SMMC-7721 tumor cells treated with two kinds cultivation by real-time quantitative polymerase chain reaction (Real-time PCR). (5) To observe changes of the tumor volume in nude mice after subcutaneous injection of SMMC-7721 tumor cells treated with two kinds of cultivation and to compare their tumorigenicity and multiplication capacity. Results Growing like typical cobblestone and arraying regularly for SMMC-7721 tumor cells in ordinary medium were observed under inverted microscope, while SMMC-7721 tumor cells in suspensive culture gathering from single cell into spherical or near spherical cells. The expression of CD90 and CD24 by the flow cytometryin suspension culture group was significantly higher than that in adherent culture group. The difference was statistically significant (CD90: P=0.000; CD24: P=0.000). But there was no significant difference between the expressions of CD133 in suspension culture group and adherent culture group. The Real-time PCR results showed that the mRNA expression of CD90 and CD24 in suspension culture group was significantly higher than that in adherent culture group. The difference was statistically significant (CD90: P=0.020, CD24: P=0.000). But there was no significant difference on expressions between suspension culture group and adherent culture group for the CD133. Statistical analysis of the tumorigenic ability and capacity to proliferate in vivo of tumor cells treated with two kinds of cultivation showed the suspension culture group’s capacity to proliferate in vivo was better than the adherent culture group. And tumorigenic ability in the suspension culture groups was higher than that in the adherent culture group. Conclusion For the human liver cancer cell line SMMC-7721 cells, hepatocarcinoma stem-like cells in suspension culture could be obtained more than that in adherent culture. Key words: Liver cancer stem-like cells; Suspension culture; SMMC-7721; CD90; CD133; CD24

Dendrobium candidum extract inhibits proliferation and induces apoptosis of liver cancer cells by inactivating Wnt/β-catenin signaling pathway

BackgroundDendrobium candidum extract (DCE) has been reported to have anti-tumor property. However, the effect of DCE on liver cancer has not been well explored. This study was aimed to evaluate the inhibitory effect of DCE on liver cancer cells. MethodsThe effect of DCE on liver cancer cells was analyzed by detecting cell viability by MTT assay, detecting colony formation ability by colony formation assay, determining apoptotic cell rate, cell cycle distribution, active caspase-3 positive cells, Ki-67 positive cells, reactive oxygen species (ROS) level, and mitochondrial transmembrane potential (MMP) by flow cytometry analysis, and analyzing changes of expressions of cell cycle-, apoptosis-, and Wnt/β-catenin pathway-related proteins by Western blot. ResultsDCE inhibited viability and promoted apoptosis of liver cancer cell lines SMMC-7721 and BEL-7404. DCE decreased colony formation, induced cell cycle arrest, and led to cell cycle-associated proteins’ abnormal expressions in SMMC-7721 and BEL-7404 cells. DCE effectively suppressed viability and proliferation of primary liver cancer cells and also induced aberrant expressions of cell cycle- and apoptosis-related proteins. After DCE treatment, ROS level was increased and MMP level was decreased. DCE inhibited β-catenin level in the nucleus and regulated downstream genes of β-catenin and further blocked Wnt/β-catenin pathway in SMMC-7721 and BEL-7404 cells as well as primary liver cancer cells. ConclusionDCE suppressed liver cancer SMMC-7721 and BEL-7404 cells as well as primary liver cancer cells likely though activating mitochondria apoptosis pathway and inducing inhibition of Wnt/β-catenin pathway.

Rhein reverses doxorubicin resistance in SMMC-7721 liver cancer cells by inhibiting energy metabolism and inducing mitochondrial permeability transition pore opening.

Rhein, a monomeric anthraquinone obtained from the plant herb species Polygonum multiflorum and P. cuspidatum, has been proposed to have anticancer activity. This activity has been suggested to be associated with mitochondrial injury due to the induction of mitochondrial permeability transition pore (mPTP) opening. In this study, the effects of 5-80 μM rhein on cell viability, half-maximal inhibitory concentration (IC50 value), resistance index, and apoptosis were assessed in the liver cancer cell lines SMMC-7721 and SMMC-7721/DOX (doxorubicin-resistant cells). Rhein (10-80 μM) significantly reduced the viability of both cell lines; 20 μM rhein significantly increased sensitivity to DOX and increased apoptosis in SMMC-7721 cells, but reversed resistance to DOX by 7.24-fold in SMMC-7721/DOX cells. Treatment with rhein increased accumulation of DOX in SMMC-7721/DOX cells, inhibited mitochondrial energy metabolism, decreased cellular ATP, and ADP levels, and altered the ratio of ATP to ADP. These effects may result from the binding of rhein with voltage-dependent ion channels (VDACs), adenine nucleotide translocase (ANT), and cyclophilin D, affecting their function and leading to the inhibition of ATP transport by VDACs and ANT. ATP synthesis was greatly reduced and mitochondrial inner membrane potential decreased. Together, these results indicate that rhein could reverse drug resistance in SMMC-7721/DOX cells by inhibiting energy metabolism and inducing mPTP opening. © 2018 BioFactors, 45(1):85-96, 2019.

Meroterpenoids isolated from Arnebia euchroma (Royle) Johnst. and their cytotoxic activity in human hepatocellular carcinoma cells

Six previously undescribed naturally occurring meroterpenoids (2, 5–9) together with seven known meroterpenoids (1, 3, 4, 10–13) were isolated from the root plant of Arnebia euchroma. Their structures and absolute configurations were determined by extensive 1D (1H NMR, 13C NMR) and 2D NMR (1H1H COSY, DEPT, HMQC, HMBC, NOESY) spectroscopic methods, spectroscopy high resolution mass spectrometry, as well as DFT and MM2 force-field calculations. Meroterpenoids 1–13 were evaluated for their cytotoxicities against human liver cancer cell lines SMMC-7721, HepG2, QGY-7703 and HepG2/ADM. Meroterpenoid 5 exhibited the most potent activity with IC50 values of 6.40 ± 0.51, 3.86 ± 0.28, 3.43 ± 0.27 and 11.31 ± 0.67 μM, respectively. Meroterpenoid 4 exhibited significant growth inhibitory effects against HepG2/ADM with IC50 at 18.77 ± 1.23 μM, and meroterpenoid 8 with IC50 at 5.41 ± 0.51 and 6.18 ± 0.47 μM against HepG2 and QGY-7703, respectively. These were more potent than the positive drug, Cisplatin.

Ent-Sauchinone as Potential Anticancer Agent Inhibiting Migration and Invasion of Human Liver Cancer Cells via Suppressing the STAT3 Signaling Pathway.

ent-Sauchinone, a lignan isolated from Saururus chinensis (Lour.) Baill., was reported that it could modulate the expression of signal transducer and activator of transcription 3 (STAT3). Since STAT3 plays a key role in invasion, migration, and metastasis of cancer, we investigated whether ent-sauchinone could exert promising inhibitory effects on the invasion and migration of the metastatic human liver cancer cell line SMMC-7721 in the present study. ent-Sauchinone was extracted from dried herbs of Saururus chinensis (Lour.) Baill. Human liver cancer cell lines SMMC-7721 and HCCLM3 were used to test the effect of ent-sauchinone on cell viability. The IC50 values and time-dependent effect of ent-sauchinone were determined by MTT assay. Cell migration and invasion of SMMC-7721 were evaluated by the wound healing test and transwell assay respectively, the known anti-metastasis agent curcumin was used as a positive control. Western blotting assay was used to investigate relevant molecular mechanisms of cell invasion and migration. Though ent-sauchinone didn't show high cytotoxicity, the wound healing assay and transwell migration assay revealed a profound impairment in the metastatic potential of SMMC-7721 cells due to down-regulation of N-cadherin, MMP-2, and MMP-9 proteins induced by inhibiting the phosphorylation of STAT3. These findings suggest that ent-sauchinone could be used as a promising agent to treat cancer metastasis.

Expression of β-catenin protein in hepatocellular carcinoma and its relationship with alpha-fetoprotein.

This study aimed to investigate the expression of β-catenin in hepatocellular carcinoma (HCC) tissues and its relationship with α-fetoprotein (AFP) in HCC. Immunohistochemistry was used to determine the expression of β-catenin in normal liver tissues (n=10), liver cirrhosis tissues (n=20), and primary HCC tissues (n=60). The relationship between β-catenin expression and clinical parameters of HCC was investigated. Real-time PCR and Western blotting were used to detect the mRNA and protein expression levels of β-catenin in the liver cancer cell line SMMC-7721 transfected with a plasmid encoding AFP, and also the mRNA and protein expression levels of β-catenin were measured in the liver cancer cell line Huh7 before and after the transfection with AFP shRNA plasmids. The results showed that β-catenin was only expressed on the cell membrane in normal liver tissues. Its localization to the cytoplasm and nucleus of cells was observed in a small proportion of cirrhotic tissues or adjacent HCC tissues, and such ectopic expression of β-catenin was predominant in HCC tissues. The abnormal expression of β-catenin was correlated with serum AFP levels, cancer cell differentiation and vascular invasion (P<0.05). Additionally, the increased expression of AFP resulted in the upregulation of β-catenin mRNA and protein levels, while knockdown of AFP with AFP shRNA led to significantly decreased β-catenin mRNA and protein levels (P<0.05). It was suggested that the abnormal expression of β-catenin is implicated in hepatic carcinogenesis and development. AFP can lead to increased expression of β-catenin, which may account for the poor prognosis of AFP-associated HCC patients.

Bovine lactoferricin P13 triggers ROS-mediated caspase-dependent apoptosis in SMMC7721 cells.

Bovine lactoferricin P13 (LfcinB-P13) is a peptide derived from LfcinB. In the present study, the effect of LfcinB-P13 on the human liver cancer cell line SMMC7721 was investigated in vitro and in vivo. The results of the present study indicate that LfcinB-P13 significantly decreased SMMC7721 cell viability in vitro (P=0.032 vs. untreated cells), while exhibiting low cytotoxicity in the wild-type liver cell line L02. In addition, the rate of apoptosis in SMMC7721 cells was significantly increased following treatment with 40 and 60 µg/ml LfcinB-P13 (P=0.0053 vs. the control group), which was associated with an increase in the level of reactive oxygen species (ROS) and the activation of caspase-3 and -9. Furthermore, ROS chelation led to the suppression of LfcinB-P13-mediated caspase-3 and -9 activation in SMMC7721 cells. LfcinB-P13 was demonstrated to markedly inhibit tumor growth in an SMMC7721-xenograft nude mouse model. The results of the present study indicate that LfcinB-P13 is a novel candidate therapeutic agent for the treatment of liver cancer.

The impact of RNA interference-induced ADAR1 down-regulation on cell proliferation of liver cancer

Objective To observe the impact of RNA interference-induced ADAR1 down-regulation on cell proliferation of liver cancer. Methods Small interfering RNA (siRNA) was transfected into liver cancer cell line SMMC-7721. ADAR1 expression was detected by RT-PCR and Western blotting. Cell proliferation was determined by methyl thiazol tetrazolium (MTT) assay. Results After transfection for 24, 48, and 72 h, ADAR1 mRNA expression was 0.612±0.086, 0.264±0.018, 0.156±0.063 in experimental group and 1.032±0.107, 0.898±0.092, 0.968±0.074 in control group, respectively. Experimental group had significantly lower ADAR1 mRNA than the other groups (P 0.05). ADAR1 protein relative expression was 0.684±0.079, 0.324±0.042, 0.145±0.058 in experimental group and 1.002±0.092, 0.917±0.068, 0.972±0.073 in control group, respectively, which was statistically significant (P<0.05). After transfection with siRNA, the proliferation ability of SMMC-7721 cells was enormously inhibited (P<0.05). Conclusion ADAR1 mRNA and protein level could be significantly decreased by specific RNA interfe-rence, and cell proliferation in SMMC-7721 cells were also greatly inhibited. Key words: RNA interference; Gene, ADAR1; Liver cancer

Lactobionic acid and carboxymethyl chitosan functionalized graphene oxide nanocomposites as targeted anticancer drug delivery systems

In this work, we report a targeted drug delivery system built by functionalizing graphene oxide (GO) with carboxymethyl chitosan (CMC), fluorescein isothiocyanate and lactobionic acid (LA). Analogous systems without LA were prepared as controls. Doxorubicin (DOX) was loaded onto the composites through adsorption. The release behavior from both the LA-functionalized and the LA-free material is markedly pH sensitive. The modified GOs have high biocompatibility with the liver cancer cell line SMMC-7721, but can induce cell death after 24h incubation if loaded with DOX. Tests with shorter (2h) incubation times were undertaken to investigate the selectivity of the GO composites: under these conditions, neither DOX-loaded system was found to be toxic to the non-cancerous L929 cell line, but the LA-containing composite showed the ability to selectively induce cell death in cancerous (SMMC-7721) cells while the LA-free analogue was inactive here also. These findings show that the modified GO materials are strong potential candidates for targeted anticancer drug delivery systems.

AP-2α inhibits hepatocellular carcinoma cell growth and migration.

Transcription factor AP-2α is involved in many types of human cancers, but its role in hepatocellular carcinogenesis is largely unknown. In this study, we found that expression of AP-2α was low in 40% of human hepatocellular cancers compared with adjacent normal tissues by immunohistochemical analysis. Moreover, AP-2α expression was low or absent in hepatocellular cancer cell lines (HepG2, Hep3B, SMMC-7721 and MHHC 97-H). Human liver cancer cell lines SMMC-7721 and Hep3B stably overexpressing AP-2α were established by lentiviral infection and puromycin screening, and the ectopic expression of AP-2α was able to inhibit hepatocellular cancer cell growth and proliferation by cell viability, MTT assay and liquid colony formation in vitro and in vivo. Furthermore, AP-2α overexpression decreased liver cancer cell migration and invasion as assessed by wound healing and Transwell assays, increasing the sensitivity of liver cancer cells to cisplatin analyzed by MTT assays. Also AP-2α overexpression suppressed the sphere formation and renewed the ability of cancer stem cells. Finally, we found that AP-2α is epigenetically modified and modulates the levels of phosphorylated extracellular signal-regulated protein kinase (ERK), β-catenin, p53, EMT, and CD133 expression in liver cancer cell lines. These results suggested that AP-2α expression is low in human hepatocellular cancers by regulating multiple signaling to affect hepatocellular cancer cell growth and migration. Therefore, AP-2α might represent a novel potential target in human hepatocellular cancer therapy.

Screening of differentially expressed genes after silencingMCM7in liver cancer cell line SMMC-7721

Screening of differentially expressed genes after silencing<i>MCM7</i>in liver cancer cell line SMMC-7721

CD147 monoclonal antibody mediated by chitosan nanoparticles loaded with α-hederin enhances antineoplastic activity and cellular uptake in liver cancer cells.

An antibody that specifically interacts with an antigen could be applied to an active targeting delivery system. In this study, CD147 antibody was coupled with α-hed chitosan nanoparticles (α-Hed-CS-NPs). α-Hed-CS-CD147-NPs were round and spherical in shape, with an average particle size of 148.23 ± 1.75 nm. The half-maximum inhibiting concentration (IC50) of α-Hed-CS-CD147-NPs in human liver cancer cell lines HepG2 and SMMC-7721 was lower than that of free α-Hed and α-Hed-CS-NPs. α-Hed-induced cell death was mainly triggered by apoptosis. The increase in intracellular accumulation of α-Hed-CS-CD147-NPs was also related to CD147-mediated internalization through the Caveolae-dependent pathway and lysosomal escape. The higher targeting antitumor efficacy of α-Hed-CS-CD147-NPs than that α-Hed-CS-NPs was attributed to its stronger fluorescence intensity in the tumor site in nude mice.

A phenylacetaldehyde–flavonoid adduct, 8-C-(E-phenylethenyl)-norartocarpetin, exhibits intrinsic apoptosis and MAPK pathways-related anticancer potential on HepG2, SMMC-7721 and QGY-7703

Norartocarpetin, quercetin and naringenin were found to effectively inhibit 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) formation through trapping its phenylacetaldehyde and form their adducts in roast beef patties. Six adducts [8-C- or 6-C-(E-phenylethenyl) flavonoids] formed between phenylacetaldehyde and three flavonoids were detected in roast beef patties by UPLC–MS analyses and compared with their synthetic references. These flavonoid–phenylacetaldehyde adducts were synthesised and further subjected to cytotoxicity tests on three liver cancer cell lines HepG2, SMMC-7721 and QGY-7703. The adduct 8-C-(E-phenylethenyl)norartocarpetin (NARA1) was found to significantly induce cancer cell death with IC50 values about 7μM. After pre-treating with MAPK and caspase inhibitors, alteration of the cell morphology and cleaved-PARP were detected in liver cancer cell lines administered with NARA1. These data indicated that norartocarpetin could inhibit PhIP formation in roast beef patties and form norartocarpetin–phenylacetaldehyde adducts. The adduct NARA1 has anticancer potential via intrinsic caspase-dependent and cell context-dependent MAPKs pathways.

First synthesis of novel 3,3′-bipyridazine derivatives as new potent antihepatocellular carcinoma agents

Hepatocellular carcinoma is one of the most common kind of cancers in clinical, and its clinical treatment is quite difficult. The latest research suggests that pyridazinone with a novel molecular skeleton shows excellent activity against hepatocellular carcinoma in vitro and in vivo. Considering YHHU-759 as the leading compound for reasonable structure modification, a series of 3,3′-bipyridazine derivatives including 8 novel compounds in this study were designed, synthesized and screened for their anti-hepatoma activities against liver cancer cell lines SMMC-7721, BEL-7402 and QGY-7703 in vitro. The result shows that all of the 3,3′-bipyridazine derivatives have good anti-hepatoma activities.