Hepatic Carcinoma Cells Research Articles

RETRACTED: Thymoquinone sensitizes human hepatocarcinoma cells to TRAIL-induced apoptosis via oxidative DNA damage

Hepatocellular carcinoma (HCC) remains one of the most predominant types of digestive system malignancies worldwide. TNF-related apoptosis-inducing ligand (TRAIL) is a biological cytokine with the mentioned specificity, but some tumor cells' resistance limits its use as a therapeutic approach. The present study aimed to investigate thymoquinone (TQ) and TRAIL's combined effect and the potential mechanisms in human hepatic HepG2 carcinoma cells. Cell viability and IC50 dose for TQ and TRAIL, alone and in combination, were determined using the MTT method. ELISA evaluated the expression levels of 8-Hydroxy-2'-deoxyguanosine. The apoptosis rate was assessed by flow cytometry, ELISA cell death assay, and caspase 8 activity assays. The mRNA and protein evaluation of candidate genes, including survivin, Bcl-2, XIAP, c-IAP1, c-IAP2, and c-FLIP, were accomplished before and after the treatment using qRT-PCR and Western blot analysis, respectively. Our results showed that TQ synergistically increased TRAIL's cell toxic effects as follows: TQ plus TRAIL > TRAIL > TQ. TQ could sensitize the HepG2 cells against the TRAIL-induced apoptosis and amplify the caspase 8 activity. This outcome is achieved by decreasing the mRNA and protein expression levels of anti-apoptotic genes. Our findings suggest that TQ can sensitize the human HCC cell line HepG2 against TRAIL by inducing the death receptor pathway. Moreover, these agents' combinational therapy might promise a therapeutic regimen for improving the clinical efficacy of TRAIL-induced apoptosis in patients with HCC.

Bluetongue Viruses Act as Novel Oncolytic Viruses to Effectively Inhibit Human Renal Cancer Cell Growth In Vitro and In Vivo.

BackgroundThe bluetongue virus (BTV) is the prototype virus in the genus Orbivirus within the family Reoviridae. Recent studies indicate that BTVs are capable of infecting and selectively lysing human hepatic carcinoma cells (Hep-3B) and prostate carcinoma cells (pc-3). This study was designed to evaluate the oncolytic potential of BTV in experimental models of human renal cancer in vitro and in vivo.Material/MethodsFive human renal cancer cell lines, ACHN, CAKI-1, OS-RC-2, 786-O, and A498, were used in this study to analyze BTV replication. These cells were lysed by oncolysis compared to normal control. Xenograft models were used to assess the efficacy and toxicity of BTVs in vivo. Data were analyzed by one-way ANOVA or two-sided unpaired t tests.ResultsThe results showed HPTEC cells to be relatively resistant to cytotoxic effects of BTVs and exhibited normal growth rate even at high dose of BTVs. Nonetheless, the renal cancer cells showed a remarkably higher sensitivity to BTVs. Moreover, the ultramicroscopic subcellular changes were also detected in the renal cells. The viral particles were observed in all the RCC cell lines, but not in HPTEC cells. Intratumoral injections of BTVs significantly decreased the tumor volume as compared to animals that received no virus treatment. Infection with BTVs significantly increased the percentage of apoptotic renal cancer cells but not the HPTEC cells. Moreover, BTV triggered apoptosis in renal cancer cells via a mitochondria-mediated pathway.ConclusionsThis study for the first time demonstrated the oncolytic potential of BTV in experimental models of human renal cancer. BTV exhibits the potential to inhibit human renal cancer cell growth in vitro and in vivo.

Effects of Curcuma comosa Roxb. Extract on the Expression of CYP450s in Immortalized Hepatocyte-like Cells (imHC)

Hepatocytes are the most abundant liver cells that produce biotransformation enzymes, including phase I metabolism enzymes and cytochrome P450s (CYP450s). These enzymes are induced or suppressed by several drugs and chemicals. Here, immortalized hepatocyte-like cells (imHC) were applied as a novel hepatocyte to study the CYP450 enzyme expression induced by a Thai folk herb, Curcuma comosa Roxb. (C. comosa). The results demonstrated that the mRNA expression levels were dependent on cell cultivation conditions, the extract's concentration, and the duration of treatment. In the 3-day cultivation, 1 µg/mL C. comosa extract upregulated the mRNA expression levels of CYP1A1, CYP1A2, CYP2B6, CYP2C9, CYP2D6, CYP2E1 and CYP3A4 in imHC, after treatment for 48 h, which was comparable to the expression levels of those genes in HepG2 cells. In addition, C. comosa increased CYP3A4 protein expression in both imHC and HepG2 cells. Furthermore, 1 µg/mLC. comosa could neutralize erythromycin's inhibitory effect, a specific CYP3A4 inhibitor, on CYP3A4 enzyme activity. In conclusion, imHC exhibited several detoxification enzymes, and this cell could be used to replace hepatic carcinoma cells in hepatotoxic studies. The function of C. comosa as a CYP enzyme-inducing agent indicates a promising treatment for the prevention of drug-induced hepatotoxicity by activating detoxification enzymes.

Nanoparticles of Punicalagin Synthesized from Pomegranate (Punica Granatum L.) with Enhanced Efficacy Against Human Hepatic Carcinoma Cells

Punicalagin is the most popular ellagitannin found in pomegranate husk and is well known to reduce the risk of cancer and cardiovascular diseases (CVDs). The present work describes a novel method for the preparation of nanoparticles of Punicalagin using pomegranate (Punica granatum) since the use of Punicalagin in bulk form is associated with many problems. In this study, nanoparticles of Punicalagin were synthesized, characterized using various techniques, and further evaluated for their antioxidant, antibacterial, and antiproliferative potential using HepG2 cancer cells. DLS analysis revealed that the average size of nanoparticles of Punicalagin was 87 nm, whereas SEM analysis confirmed spherical shaped nanoparticles with size ranging from 90 to 116 nm. HPLC studies confirmed the presence of Punicalagin in synthesized nanoparticles. The results also revealed that nanoparticles of Punicalagin were nearly four times more potent antioxidants than bulk and possessed an inhibition zone of about 13 mm. The response of antiproliferative assay showed that the nanoparticles of Punicalagin caused nearly 44% reduction while bulk form showed only a 15% reduction in cell viability of cancerous cells at 100 µg/mL. The study suggests a great potential for use of the herbal drug in nano form to treat cancer as compared to its bulk counterparts.

Thiolated polymer nanocarrier reinforced with glycyrrhetinic acid for targeted delivery of 5-fluorouracil in hepatocellular carcinoma.

The present work investigates the targeting efficacy of a novel thiolated polymer-based nanocomposite reinforced with glycyrrhetinic acid (GA) and loaded with 5-fluorouracil in hepatocellular carcinoma (HCC). The thiolated polymers were synthesized by EDAC-mediated conjugation reactions and lyophilization. The nanoparticles were prepared by solvent diffusion and high-pressure homogenization method. The prepared nanocomposite was characterized by Fourier transform infrared (FTIR) radiation, x-ray diffraction (XRD), dynamic light scattering (DLS), scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. Pharmacological evaluation of the formulation was carried out on a rat model of diethylnitrosamine (DEN), and carbon tetrachloride (CCl4)-induced HCC and MTT assay was carried out with HEP-G2 cell line. In silico studies were conducted to investigate the probable mechanistic pathway of the nanocomposite. FTIR and XRD analysis indicated the successful thiolation of the polymers and confirmed the formation of the nanocomposite without any incompatibilities. DLS, SEM/EDX and AFM characterization confirmed that the nanoparticles were within the nano-size range. MTT assay implied the cytotoxic nature of the nanocomposite against hepatic carcinoma cells. The in vivo study revealed that serum SGOT, SGPT, ALP, GGT and total bilirubin levels were significantly reduced, in comparison with disease control and the result was confirmed by histopathology studies. The results of the HPLC analysis of liver homogenate confirmed the liver targeting ability of the nanocomposite. In silico studies exhibited significant binding affinity of GA and thiolated Eudragit towards liver homolog receptor-1 (LRH-1) suggesting that the developed nanocomposite could be a potential material for the treatment of HCC.

Evaluation of free radical scavenging with in vitro antiproliferative properties of different extracts of pluchea lanceolata (dc.) oliv. and hiern in cancer cell lines

Background: Traditional medicinal plants have gained attention as a repository of pharmacologically active molecules. Extracts derived from Pluchea lanceolata (DC.) Oliv. and Hiern are reported to be antimalarial and can protect against chemical-induced neurotoxicity. There is limited research on solvents for extraction of metabolites from stem of P. lanceolata and their anti-cancer potential, which needs to be investigated. Objectives: The objective of the study was to investigate potency of stem powder, extracted in ethanol, methanol, aqueous, and phosphate buffer saline (PBS) solvents, for their phytochemical content, antioxidant potential, and in vitro antiproliferative nature in human cancer cell lines. Materials and Methods: The stem extracts of P. lanceolata were evaluated by phytochemical assays, antioxidant assays (2,2-diphenyl-1-picrylhydrazyl [DPPH] radical scavenging assay, hydrogen peroxide radical scavenging assay, nitric oxide radical scavenging assay, total antioxidant capacity, and assay of reducing power), and antiproliferative potential against cervical (ME-180 and HeLa) and hepatic (HepG2) carcinoma cell lines by MTT assay. Results: Quantification studies showed that the total phenolic content was in the range 7.44–38.91 mg GAE/g of stem extract, while the flavonoids were present in the range 29.05–109.62 mg QE/g of stem extract. Aqueous (DPPH antioxidant capacity assay±PVPP, H2O2 free radical scavenging method-PVPP, assay of reducing power+PVPP, and total antioxidant capacity-PVPP), methanol (H2O2 free radical scavenging method+PVPP, NO radical scavenging assay+PVPP, total antioxidant capacity-PVPP), and ethanol (NO radical scavenging assay-PVPP, assay of reducing power-PVPP) extracts had the highest antioxidant potential in respective assays. MTT findings demonstrated that the aqueous extract was more potent in ME-180 and HepG2 cell lines while the PBS extract caused maximal cytotoxicity in HeLa cells. HepG2 cells were more susceptible than ME-180 and HeLa cells for any of the extract or standard drug evaluated. Conclusion: Aqueous extract of P. lanceolata stem is the most promising extract for further cancer-cell toxicity.

Anticancer effect of myristicin on hepatic carcinoma and related molecular mechanism

Context Myristicin is a natural active compound that has inflammatory, antimicrobial and anti-proliferative properties. Yet, its effect on hepatic carcinoma has not been investigated. Objective To explore the role and related molecular mechanism of myristicin in hepatic carcinoma in vitro. Materials and methods Human hepatic carcinoma cell lines (Huh-7 and HCCLM3 cells) were treated with different concentrations of myristicin (0.5, 1 and 5 mM) for 24, 48 and 72 h. Then, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium assay (MTT), flow cytometer (FCM) analysis and transwell assay were performed to determine cell proliferation, apoptosis and migration/invasion, respectively. Protein levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X (Bax), E-cadherin, N-cadherin and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signalling pathway-related proteins were detected using Western blot assay. Gene expression was determined using quantitative real time-polymerase chain reaction (qRT-PCR). Results Myristicin inhibited cell proliferation and induced apoptosis in Huh-7 and HCCLM3 cells; suppressed cell migration and invasion ability, and increased E-cadherin expression and decreased N-cadherin expression, thereby inhibiting epithelial–mesenchymal transition (EMT). Finally, the findings indicated that myristicin decreased phosphorylated (p)-mTOR and p-AKT expression at the protein level. Discussion and conclusions Myristicin exerts an efficient therapeutic effect on hepatic carcinoma by suppressing PI3K/Akt/mTOR signalling pathway; thus, it may be used as a new potential drug for hepatic carcinoma treatment.

The Long Noncoding RNA LOXL1-AS1 Promotes the Proliferation, Migration, and Invasion in Hepatocellular Carcinoma.

Objective To investigate the expression of long noncoding RNA lysyl oxidase-like 1-antisense 1 (LOXL1-AS1) in hepatocellular carcinoma tissues and its effect on cell proliferation, migration, and invasion. Methods Quantitative real-time PCR was used to analyze the expression of LOXL1-AS1 RNA in tumor tissues, adjacent normal tissues, and cell lines. MTT assay, colony formation assay, flow cytometry analysis, transwell assays, and lentivirus-mediated RNA interference (RNAi) technology were used to evaluate cell proliferation and migration. Results In the present study, we observed that the expression level of LOXL1-AS1 in hepatocellular carcinoma tissue was significantly higher than that in adjacent nontumor tissues, and its expression in three hepatic carcinoma cell lines was obviously higher than that in a normal cell line. In addition, in the Hep-G2 cell line, LOXL1-AS1 downregulation significantly inhibited cell proliferation in the light of the MTT and colony formation assays in vitro, which was consistent with animal experiment in vivo. What is more, cell migration was also inhibited in vitro in Matrigel Transwell Assay by LOXL1-AS1 knockdown, which might be partly attributed to the reduction of MMP-2 and MMP-9 protein expressions. Finally, cell cycle analysis revealed that knockdown of LOXL1-AS1 induced significantly a G0/G1 phase cell cycle arrest, which might be partly attributed to the downregulation of Cdc2, Cdc25A, and cyclin B1 protein expression. Conclusion In conclusion, we demonstrated that reduced LOXL1-AS1 expression could inhibit hepatocellular carcinoma cell proliferation, migration, and invasion. The application of RNAi targeting LOXL1-AS1 might be a potential treatment strategy in advanced cases.

Hyperspectral microscopy combined with DAPI staining for the identification of hepatic carcinoma cells.

In this study, the DAPI staining is firstly reported for use in the identification of hepatic carcinoma cells based on hyperspectral microscopy. Nuclei in cancer cells usually contain more aneuploidies than that in normal cells, leading to the change of DNA content. Here, we stain hepatic carcinoma tissues and normal hepatic tissues with 4',6-diamidino-2-phenylindole (DAPI) which is sensitive to the DNA content as a fluorochrome binds to DNA. Consequently, the difference in DNA content between hepatic carcinoma cells and normal hepatic cells can be identified by the fluorescent spectral characteristics. Harnessing the hyperspectral microscopy, we find that the fluorescent properties of these two kinds of cells are different not only in the intensity but also in the spectral shape. These properties are exploited to train a support vector machine (SVM) model for classifying cells. The results show that the sensitivity and specificity for the identification of 1000 hepatic carcinoma samples are 99.3% and 99.1%, respectively.

Euchromatin histone methyltransferase II (EHMT2) regulates the expression of ras-related GTP binding C (RRAGC) protein

Dimethylation of the histone H3 protein at lysine residue 9 (H3K9) is mediated by euchromatin histone methyltransferase II (EHMT2) and results in transcriptional repression of target genes. Recently, chemical inhibition of EHMT2 was shown to induce various physiological outcomes, including endoplasmic reticulum stress-associated genes transcription in cancer cells. To identify genes that are transcriptionally repressed by EHMT2 during apoptosis, and cell stress responses, we screened genes that are upregulated by BIX-01294, a chemical inhibitor of EHMT2. RNA sequencing analyses revealed 77 genes that were upregulated by BIX-01294 in all four hepatic cell carcinoma (HCC) cell lines. These included genes that have been impli-cated in apoptosis, the unfolded protein response (UPR), and others. Among these genes, the one encoding the stress-response protein Ras-related GTPase C (RRAGC) was upregulated in all BIX-01294-treated HCC cell lines. We confirmed the regulatory roles of EHMT2 in RRAGC expression in HCC cell lines using proteomic analyses, chromatin immune precipitation (ChIP) assay, and small guide RNA-mediated loss-of-function experiments. Upregulation of RRAGC was limited by the reactive oxygen species (ROS) scavenger N-acetyl cysteine (NAC), suggesting that ROS are involved in EHMT2-mediated transcriptional regulation of stress-response genes in HCC cells. Finally, combined treatment of cells with BIX-01294 and 5-Aza-cytidine induced greater upregulation of RRAGC protein expression. These findings suggest that EHMT2 suppresses expression of the RRAGC gene in a ROS-dependent manner and imply that EHMT2 is a key regulator of stress-responsive gene expression in liver cancer cells.

Green Synthesis of AgNPs() Ultilizing Delonix Regia Extract as Anticancer and Antimicrobial Agents**

AbstractSilver is an effective antimicrobial agent with low toxicity, which is important especially in the treatment of burn wounds where transient bacteremia is prevalent. In this research, the green method was employed to synthesis silver nanoparticles (AgNPs). The synthesized AgNPs were characterized using IR, XRD, DLS and UV/Visible Spectrophotometery. XRD pattern of AgNPs has a face‐centered cubic (FCC) form and crystalline lattice with a crystallite size of 12 nm. FT‐IR spectra indicate the functional groups of phytochemical compounds at Delonix regia extract DRE and AgNPs. DLS showed the distribution of average particle size of AgNPs that have 11 nm mean size of the particle. The prepared AgNPs were screened for their cytotoxic effect against colon carcinoma cells (HCT‐ 116 cell line), hepatic cellular carcinoma cells (HepG‐2) and breast carcinoma cells (MCF‐7). The cytotoxic effect of AgNPs is close to the cytotoxic effect of Doxorubicin standard drug. The toxicity of the AgNPs was tested on bacterial species such as Gm (+) positive bacteria (Bacillus subtilis) and Gm (‐) negative bacteria (Serrati amarcescence and Escherichia coli) and fungal species such as Candida albicans, Getrichm candidum and Aspergillus flavus. The antimicrobial activity of AgNPs was greater towards Gm (+) positive bacteria compared to Gm (‐) negative bacteria. These biological activities of the synthesized AgNPs could be promised to use it as a drug.

LncRNA UCA1 Affects the Cell Proliferation, Migration, Invasion and Apoptosis of Hepatic Carcinoma Cells by Targeting MicroRNA-193a-3p.

Objective/BackgroundHepatic carcinoma (HCC) is the fourth lethal cancer in the world, but its relationship with lncRNA urothelial cancer-associated 1 (UCA1)/microRNA-193a-3p axis remains unclear, so this study would explore the relationship.MethodsA real-time polymerase chain reaction (RT-PCR) assay was carried out to quantify lncRNA UCA1 and microRNA-193a-3p in HCC tissues and cells, and relevant overexpression or inhibition vectors were constructed to analyze the influences of lncRNA UCA1 and microRNA-193a-3p on HCC cells. A Transwell assay was used to measure invasion and migration of HCC cells, and a Western blot assay to quantify protein biomarkers of apoptosis, invasion, and migration, a MTT assay to determine cell viability, a flow cytometry to detect cell cycle, and a dual-luciferase reporter gene assay to analyze the correlation between lncRNA UCA1 and microRNA-193a-3p.ResultsLncRNA UCA1 was increased in HCC, while microRNA-193a-3p was decreased. Down-regulated lncRNA UCA1 could up-regulate microRNA-193a-3p, and down-regulated lncRNA UCA1 or up-regulated microRNA-193a-3p would strengthen cell apoptosis and weaken cell migration, invasion, and proliferation. Furthermore, lncRNA UCA1 could negatively regulate microRNA-193a-3p by binding to it.ConclusionLncRNA UCA1 promotes malignant hyperproliferation of HCC cells by repressing microRNA-193a-3p.

Enhanced targeted delivery of adenine to hepatocellular carcinoma using glycyrrhetinic acid-functionalized nanoparticles in vivo and in vitro

Hepatocellular carcinoma (HCC), a common malignancy in China and globally, is primarily treated through surgical resection and liver transplantation, with chemotherapy as a significant synergistic option. Adenine (Ade), a nucleobase, exhibits antitumor effects by blocking human hepatic carcinoma cells in S phase and inhibiting tumor cell proliferation. However, its use is limited owing to its low solubility, poor targeting ability, and nephrotoxicity. Therefore, liver-targeting drug delivery systems have attracted considerable attention for the treatment of HCC. In this study, we explored the liver-targeting efficacy and antitumor effect of adenine-loaded glycyrrhetinic acid-modified hyaluronic acid (Ade/GA-HA) nanoparticles in vitro and in vivo. The GA-HA nanoparticles possessed obvious targeting specificity toward liver cancer cells, which was mainly achieved by the specific binding of the GA ligand to the GA receptor that was highly expressed on the liver cell membrane. In vitro and in vivo results showed that Ade/GA-HA nanoparticles could inhibit liver cancer cell proliferation and migration, promote apoptosis, and significantly inhibit the growth of tumor tissues. Altogether, this study is the first to successfully demonstrate that the targeting activity and antitumor effect of Ade against HCC are enhanced by using GA-HA nanoparticles in vitro and in vivo.

Identification of the DNA Replication Regulator MCM Complex Expression and Prognostic Significance in Hepatic Carcinoma.

Background The microliposome maintenance (MCM) complex, MCM2-7, is revealed to be involved in multiple cellular processes and plays a key role in the development and progression of human cancers. However, the MCM complex remains poorly elaborated in hepatic carcinoma (HCC). Methods In the study, we found the mRNA and protein level by bioinformatics. We also explored the prognostic value, genetic alteration, interaction network, and functional enrichment of MCM2-7. The MCM expression and correlation among these MCMs in HCC cell lines were identified by western blot. Results MCM2-7 was significantly increased in HCC tissues compared to normal liver tissues. The high level of MCM2-7 had a positive correlation with poor prognosis. However, MCM2-7 alterations were not correlated with poor OS. MCMs were both increased in HCC cell lines compared to the normal hepatocyte cell line. Furthermore, the positive correlation was found among MCMs in HCC cell lines. Conclusions The MCM complex was increased in HCC tissues and cell lines and negatively correlated with prognosis, which might be important biomarkers for HCC.

Bactris guineensis (Arecaceae) extract: Polyphenol characterization, antioxidant capacity and cytotoxicity against cancer cell lines

BACKGROUND: Lower risk of digestive tract cancer development has been associated with polyphenol intake. Bactris guineensis is an edible endemic palm that grows in Central and South America. OBJECTIVE: This study performs a phenolic characterization of Bactris guineensis and evaluates the bioactivity of this fruit. METHODS: The phenolic compounds of B. guineensis were characterized by HPLC-UV-HRMS analyses and the antioxidant activity was measured by chemical and cellular methods. Additionally, cytotoxicity of B. guineensis polyphenols was performed on 4 cancer cell lines and the pro-apoptotic effect was evaluated by flow cytometry using annexin staining. RESULTS: The major phenolic compounds of B. guineensis were proanthocyanidins. The extract IC50 for DPPH was 3.3±0.2 μg/mL and for induced intracellular ROS was 153±13 μg/mL. MTT cytotoxic assays demonstrate IC50 values between 16.6 and 24.9 μg/mL for the colon and hepatic adenocarcinomas, with high selectivity effects towards cancer cells compared to non-tumor cells. A 20 to 50% early apoptotic effect was observed in cancer cells lines by Annexin/PI staining. CONCLUSIONS: B. guineensis evidenced an important radical scavenging activity and a strong cytotoxic activity against hepatic and colorectal carcinoma cells, showing better values than procyanidin extracts from other fruits previously described.

MiR-145 Regulates the Chemoresistance of Hepatic Carcinoma Cells Against 5-Fluorouracil by Targeting Toll-Like Receptor 4.

Background5-fluorouracil (5-FU) is a common drug for hepatic carcinoma (HCC), but the drug resistance of clinical chemotherapy restricts its use. Studies have demonstrated that miRNA molecules can act as a chemoresistance regulator in drug resistance of tumors, whereas the role of miR-145 in the 5-FU-resistant HCC remains unclear.ObjectiveTo explore the prognostic value of miR-145 in HCC and its molecular mechanism in 5-FU-resistant HCC cells.MethodsA qRT-PCR assay was conducted to quantify miR-145 in HCC tissues and 5-FU-resistant HCC cells. The Cell Counting Kit-8 (CCK-8) and flow cytometry were adopted to analyze the proliferation and apoptosis of 5-FU-resistant HCC cells. The Western blot was adopted to quantify toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and apoptosis-related proteins. Moreover, an in vivo tumor xenotransplantation of nude mice was conducted to determine the effect of miR-145 on 5-FU-resistant HCC cells.ResultsMiR-145 was expressed lowly in HCC tissues and cells, and linked to high TNM staging and lymph node metastasis of HCC patients. Down-regulation of miR-145 indicated a poorer prognosis and it promoted drug resistance of HCC cells and inhibited cell apoptosis. In contrast, miR-145 overexpression improved the sensitivity of HCC cells to 5-FU and enhanced the inhibition of 5-FU on tumor growth. The luciferase reporter gene assay showed that TLR4 was the direct target of miR-145, and the Western blot assay revealed that overexpression of TLR4 reversed the inhibitory effect of miR-145 overexpression on TLR4 and MyD88 protein and the effects of it on apoptosis-related proteins.ConclusionMiR-145 is an inhibiting factor in HCC and can target TLR4 to mediate the chemoresistance of HCC, which may provide novel ideas for treating HCC.

Tanshinone I regulates autophagic signaling via the activation of AMP-activated protein kinase in cancer cells.

Tanshinone I, one of the components of Salvia miltiorrhiza Bunge, exhibits anti-tumor ability and induces autophagy. But the mechanisms are not fully understood. This study aims to investigate whether AMP-activated protein kinase dependent pathway is involved in the autophagic signaling regulation and its relationship with tumor suppression. Breast cancer cells (MDA-MB-231, MCF-7) and hepatocellular carcinoma cells (HepG2) were treated with Tanshinone I or vehicle. Acridine orange dyeing and transmission electron microscopy were employed to evaluate autophagic cells. MTT and Cell Counting Kit-8 assays were used to detect the effect of Tanshinone I combined with autophagy inhibitors on cell proliferation. Western blot was used to detect the expression levels of Beclin1 and LC3-I/II, as well as the phosphorylation of AMPKα and ULK1. Our results showed that Tanshinone I suppressed proliferation of HepG2, MDA-MB-231 and MCF-7 cancer cell lines. LC3-II and P62 were induced by Tanshinone I in all three cancer cell lines. But autophagic flux analysis showed that Tanshinone I treatment induced autophagy only in MDA-MB-231, which was also proved by transmission electron microscopy. Tanshinone I upregulated the phosphorylation of AMPKα and its downstream ULK1. AMP-activated protein kinase inhibitor compound C attenuated Beclin 1 and LC3-II expression induced by Tanshinone I in HepG2. In MDA-MB-231, compound C surprisingly induced LC3-II upregulation which is independent of AMPKα activation. Under this circumstance, treatment of Tanshinone I combined with compound C significantly inhibited MDA-MB-231 proliferation, compared with Tanshinone I treatment alone. This study demonstrates that Tanshinone I could induce cancer cell death and regulate autophagy signaling in breast cancer and hepatic carcinoma cells. Activation of AMPKα was found to be involved in autophagic signaling regulation by Tanshinone I.

Ultrasound cavitation enhanced chemotherapy: In vivo research and clinical application.

The novelty of this research is that we used ultrasound cavitation to enhance the effects of chemotherapy in the subcutaneous and orthotopic hepatic carcinomas in nude mice. Case reports of the effects of the targeting ultrasound cavitation and chemotherapy on malignant tumors in clinical patients were also examined. We found that low-frequency ultrasound cavitation combined with chemotherapy is effective in the inhibition of tumor growth to some extent.

Protective potency of ascorbic acid supplementation against cytotoxicity and DNA fragmentation induced by triphenyltin on human liver carcinoma cells.

Agrochemicals are one the most significant sources of environmental pollution. Cytotoxicity and genotoxicity are the serious side effects of fungicide. In the current study, I have evaluated acute cytotoxicity and genotoxicity of triphenyltin (TPT) on human hepatic carcinoma (HepG2) cells and the ameliorating effect of ascorbic acid for 24h. In this experiment, I have exposed HepG2 cells to ascorbic acids (50, 100, and 200μM) simultaneously and 24h prior triphenyltin (TPT, 400ng/ml) exposure for 24h to determine the protective effect of ascorbic acid by using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and NRU (neutral red uptake) assays. Oxidative stress, such as intracellular reactive oxygen species and glutathione levels, was measured in HepG2 cells. The intracellular reactive oxygen species was evaluated using fluorescent probe DCFDA (6-carboxy-2',7' dichloro-dihydrofluorescein diacetate). Apoptosis and genotoxicity effects of TPT in HepG2 cells were determined using flow cytometry and comet assay. The result of these experiments showed that the TPT compound (400ng/ml) induced cytotoxicity, oxidative stress and apoptosis, and DNA damage in HepG2 cells.Ascorbic acid reduced cytotoxicity, oxidative stress, apoptosis, and genotoxicity induced by TPT. Thus, ascorbic acid is a potent antioxidant, and it showed a significant protective effect against toxicity induced by TPT in HepG2 cells.

A multi-scale systems pharmacology approach uncovers the anti-cancer molecular mechanism of Ixabepilone

It has been realized that FDA approved drugs may have more molecular targets than is commonly thought. Thus, to find the exact drug-target interactions (DTIs) is of great significance for exploring the new molecular mechanism of drugs. Here, we developed a multi-scale system pharmacology (MSSP) method for the large-scale prediction of DTIs. We used MSSP to integrate drug-related and target-related data from multiple levels, the network structural data formed by known drug-target relationships for predicting likely unknown DTIs. Prediction results revealed that Ixabepilone, an epothilone B analog for treating breast cancer patients, may target Bcl-2, an oncogene that contributes to tumor progression and therapy resistance by inhibiting apoptosis. Furthermore, we demonstrated that Ixabepilone could bind with Bcl-2 and decrease its protein expression in breast cancer cells. The down-regulation of Bcl-2 by Ixabepilone is resulted from promoting its degradation by affecting p-Bcl-2. We further found that Ixabepilone could induce autophagy by releasing Beclin1 from Beclin1/Bcl-2 complex. Inhibition of autophagy by knockdown of Beclin1 or pharmacological inhibitor augmented apoptosis, thus enhancing the antitumor efficacy of Ixabepilone against breast cancer cells in vitro and in vivo. In addition, Ixabepilone also decreases Bcl-2 protein expression and induces cytoprotective autophagy in human hepatic carcinoma and glioma cells. In conclusion, this study not only provides a feasible and alternative way exploring new molecular mechanisms of drugs by combing computation DTI prediction, but also reveals an effective strategy to reinforce the antitumor efficacy of Ixabepilone.