Human Liver Cancer SMMC-7721 Cells Research Articles

Alisol B 23-Acetate Increases the Antitumor Effect of Bufalin on Liver Cancer through Inactivating Wnt/β-Catenin Axis.

Objective Liver cancer seriously threatens the health of people. Meanwhile, it has been reported that bufalin could act as an inhibitor in liver cancer. In addition, alisol B 23-acetate is a natural product derived from Alisma plantago-aquatica Linn which has an antitumor effect. In this study, we aimed to explore whether alisol B 23-acetate could increase the antitumor effect of bufalin on liver cancer. Methods In order to detect the effect of alisol B 23-acetate in combination with bufalin on liver cancer, human liver cancer SMMC-7721 and MHCC97 cells were used as subjects. Bufalin and alisol B 23-acetate were performed on cells. Cell viability was tested by MTT assay. In addition, flow cytometry was performed to assess the cell apoptosis. Autophagy-related protein levels were tested by western blotting. Results The data revealed that bufalin significantly decreased the viability of liver cancer cells, and the inhibitory effect was further increased by alisol B 23-acetate. In addition, alisol B 23-acetate notably enhanced the apoptotic effect of bufalin on liver cancer cells through mediation of Mcl-1, Bax, Bcl-2, and cleaved caspase-3. Meanwhile, alisol B 23-acetate in combination with bufalin induced the autophagy in liver cancer cells through mediation of Beclin-1 and p62. Furthermore, alisol B 23-acetate in combination with bufalin significantly downregulated the level of GSK-3β and increased the expression of β-catenin in liver cancer cells. Conclusion In summary, these findings provide the first evidence that alisol B 23-acetate improves the anticancer activity of bufalin on liver cancer through activation of the Wnt/β-catenin axis, and these outcomes might shed new lights on exploring the new methods against liver cancer.

Design, synthesis and antitumor evaluation of novel 1H-indole-2-carboxylic acid derivatives targeting 14-3-3η protein

In this work, a series of novel 1H-indole-2-carboxylic acid derivatives targeting 14-3-3η protein were designed and synthesized for treatment of liver cancer. After structural optimization for several rounds, C11 displayed a relatively better affinity with 14-3-3η, as well as the best inhibitory activities against several typical human liver cancer cell lines, including Bel-7402, SMMC-7721, SNU-387, Hep G2 and Hep 3B cells. Compound C11 also displayed best inhibitory activity against chemotherapy-resistant Bel-7402/5-Fu cells. Besides, C11 was rather safe against hERG and possessed moderate T1/2 and CL values in liver microsomes. In anti-proliferation, trans-well and cell apoptosis assays, C11 also showed its huge potential as a potent antitumor agent. Then, Western blot assay was conducted, following analyzed by molecular docking, the anti-proliferative mechanisms of this small-molecule inhibitor were revealed. Moreover, C11 was demonstrated to induce G1-S phase cell cycle arrest in liver cancer cells.

Effect of berbamine on invasion and metastasis of human liver cancer SMMC-7721 cells and its possible mechanism.

Berbamine is a bisbenzylisoquinoline alkaloid extracted from Berberis poiretii of Berberis of Berberidaceae. It has been reported that it can significantly inhibit the proliferation of a variety of malignant tumor cells, including liver cancer. However, the effect of berbamine on the invasion and metastasis of liver cancer has not been reported. The present study demonstrated that berbamine inhibited the migration and invasion of SMMC-7721 cells in a concentration-dependent manner and obviously increased the gap junction function and the expression of Cx32 in SMMC-7721 cells compared with control group. However, after silencing Cx32, berbamine had no significant effect on cell invasion and metastasis. Before silencing Cx32, the expression of PI3K and P-AKT were decreased after berbamine treated on SMMC-7721 cells for 24 h. After silencing Cx32, the expression of PI3K and P-AKT were increased in SMMC-7721 cells. The expression of PI3K and P-AKT had no significant effect after berbamine treated on SMMC-7721 cells for 24 h with silencing Cx32. In conclusion, the results of the present study suggest that berbamine could inhibit the SMMC-7721 cell migration and invasion, and its mechanism may be related to the regulation of PI3K/AKT signaling pathway by enhancing the expression of Cx32.

Study of zinc oxide nanoparticles on epithelial-mesenchymal transition and toxicity of spontaneous monocyte-mediated cytotoxity-7721 cells

To explore the method of studying zinc oxide nanoparticles on the epithelial-mesenchymal transition and toxicity of liver cancer SMMC-7721 cells. Human liver cancer SMMC-7721 cells is stored in 90% FBS+10% DMSO liquid nitrogen. ZnO suspension was prepared, cell viability was assessed using MTT assay, cell apoptosis was analyzed by flow cytometry, and EMT-related proteins were detected by western blotting. Results showed LDH activity increased continuously with the increase in ZnO nanoparticle concentration and exposure time (P < 0.001). Both ATP and SOD activities gradually decreased with the increase in ZnO nanoparticle concentration and exposure time (both P < 0.001). The MTT assay revealed that with the increase in ZnO dose, the proliferation of SMMC-7721 liver cancer cells decreased gradually (P < 0.001), and with the continuous increase in exposure time to ZnO nanoparticles, the reproductive viability of these cells also continued to decline (P < 0.001). The apoptosis rate of SMMC-7721 liver cancer cells increased with the increase in ZnO dose (P < 0.001). Flow cytometry results demonstrated that the apoptosis rate of SMMC-7721 liver cancer cells increased with the continuous prolongation of treatment time (P < 0.001). Western blotting experiments revealed that the concentrations of vimentin, Snail, N-cadherin, and Slugn proteins in SMMC-7721 cells increased significantly, whereas those of E-cadherin and ZO-1 decreased significantly, with the increase in ZnO dose (both P < 0.001). Therefore, ZnO nanoparticles can induce apoptosis of SMMC-7721 liver cancer cells, inhibit cell proliferation and EMT, and can be used as a new nanoparticle carrier for potential treatment of liver cancer.

Recognition-Driven Remodeling of Dual-Split Aptamer Triggering In Situ Hybridization Chain Reaction for Activatable and Autonomous Identification of Cancer Cells.

Proximity-dependent hybridization chain reaction (HCR) has shown great potential in sensing biomolecules on the cell surface. However, the requirement of two adjacent bioevents occurring simultaneously limits its application. To solve the problem, split aptamers with target binding ability were introduced to combine with split triggers for initiating HCR, thus producing a novel dual-split aptamer probe (DSAP). By employing cancer-related receptors as models, in situ HCR on a cancer cell surface induced by recognition-driven remodeling of the DSAP was demonstrated. The DSAP consisted of two sequences. Each contained two segments; one derived from split aptamers and the other originated in split triggers. In the presence of target cells, split aptamers reassembled on the cell surface under the "induced-fit effect", thus forcing two split triggers close to each other. The remodeled DSAP worked as an intact trigger, which opened the H1 hairpin probe and then hybridized with the H2 hairpin probe, thus initiating HCR to produce an activated fluorescence signal. As a proof of concept, human liver cancer SMMC-7721 cells and their split ZY11 aptamer were used to construct the DSAP. Results indicated that the DSAP realized sensitive analysis of target cells, permitting the actual detection of 20 cells in the buffer. Moreover, the specific identification of target cells in mixed cell samples and the quantitative analysis of target cells in serum were also achieved. The DSAP strategy is facile and universal, which not only would expand the application range of HCR but also might be developed as a multitarget detection technique for bioanalysis.

Essential oil from <i>Chenopodium ambrosioides L.</i> induces mitochondrial-mediated pathway and endoplasmic reticulum stress-related apoptosis in human liver cancer SMMC-7721 cells

Purpose: To evaluate the cytotoxic effect of essential oil derived from Chenopodium ambrosioides L. in Sichuan Province on human liver cancer SMMC-7721 cells, as well as its possible molecular mechanisms.Methods: Cytotoxicity was characterized by MTT assay and transmission electron microscopy (TEM) of SMMC-7721 cells ultrastructure. The apoptotic effect of the essential oil was evaluated by changes in mitochondrial membrane potential and Western blot assay.Results: MTT assay data indicate that the essential oil was cytotoxic to SMMC-7721 cells, while TEN revealed that there were vacuoles and nucleus fragmentation in the SMMC-7721 cell cytosol, cell swelling, and a large amount of leakage. Mitochondrial membrane potential assay and Western Blot data indicate that the essential oil induced cell apoptosis.Conclusion: The essential oil of Chenopodium ambrosioides L. in Sichuan Province seems to induce apoptosis of human liver cancer SMMC-7721 cells via the mitochondrial-mediated pathway and endoplasmic reticulum stress. Thus, this plant requires further investigation as a potential source of ananti-liver cancer drug.
 Keywords: Chenopodium ambrosioides Essential oil; Anti-tumor activity, Liver cancer Apoptosis, SMMC-7721 cells

Pure paclitaxel nanoparticles: preparation, characterization, and antitumor effect for human liver cancer SMMC-7721 cells.

IntroductionPure paclitaxel nanoparticles (PPN), consisting entirely of drug molecules, were prepared by the electrostatic spraying method as promising candidates for antitumor application. Compared with the traditional preparation method, the advantage of the electrostatic spraying method included high production rates, relatively small particle sizes, and ease of preparation.Materials and methodsPaclitaxel was used to prepared PPN by electrostatic spray. The electrostatic spray device included a constant speed pump with a syringe, a high-voltage power supply, and a metal foil receiver was used to prepare and evaluate PPN. The syringe drew off a certain amount of paclitaxel chloroform solution (150 μg/mL) and was placed on the constant speed injection pump. The dissolution behavior of PPN was evaluated by dissolution test and the presence of paclitaxel in PPN was detected by X-Ray powder diffraction and differential scanning calorimetry. Effect of PPN on SMMC-7721 cells were studied by cell uptake, cell apoptosis and antitumor study.ResultsThe results of X-ray powder diffraction and differential scanning calorimetry characterization showed that the PPN were in an amorphous state. A dissolution study indicated that PPN have a significantly enhanced dissolution rate of paclitaxel. Moreover, SMMC-7721 tumor cells treated with PPN exhibited a distinctly high uptake rate that promoted cell apoptosis. An in vivo antitumor study demonstrated that PPN had significant antitumor efficacy.ConclusionAll conclusions verified that electrostatic spraying is a potential technology for developing PPN, and PPN can be regarded as a promising treatment for cancer.

Luteolin Promotes Cell Apoptosis by Inducing Autophagy in Hepatocellular Carcinoma

Background/Aims: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is a leading cause of cancer-related death worldwide. Luteolin, a flavonoid from traditional Chinese medicine, shows anti-cancer activity in many cancer cells, including HCC. However, the mechanism underlying the action of luteolin in HCC, especially its role in regulating cell autophagy, remains unclear. In the present study, we investigated the role of luteolin in regulating cell autophagy and the role of autophagy in luteolin-induced apoptosis. Methods: The 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay (MTT) was used to investigate cell viability. Flow cytometry analysis was used to detect the cell cycle and cell apoptosis. Hoechst 33342 staining was used to detect cell apoptosis. Transmission electron microscopy was used to investigate autophagy. qRT-PCR and western blotting were used to detect apoptosis- and autophagy-related mRNAs and proteins. Results: Luteolin reduced the viability of SMMC-7721 cells in a time and dose-dependent manner, and induced significant G0/G1-phase arrest. In addition, the results of flow cytometry analysis and Hoechst 33342 staining showed that luteolin treatment increased the number of apoptotic cells obviously, and the results of qRT-PCR and western blotting showed that luteolin treatment increased caspase 8 and decreased bcl-2 at the mRNA and protein levels. Furthermore, luteolin increased the number of intracellular autophagosomes, promoted LC3B-I conversion to LC3B-II, and increased Beclin 1 expression. Finally, co-treatment with the autophagy inhibitor chloroquine weakened the effects of luteolin on cell apoptosis. Conclusion: Luteolin induced apoptosis in human liver cancer SMMC-7721 cells, partially via autophagy. Thus, luteolin could be used as a regulator of autophagy in HCC treatment.

Clinical effect of ultrasound-guided injection of biodegradable poly(lactic-co-glycolic acid)-Fe3O4 in situ implant for magnetic thermal ablation in treatment of nude mice with human liver cancer SMMC-7721 cells

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

NAT10 regulates p53 activation through acetylating p53 at K120 and ubiquitinating Mdm2.

As a genome guardian, p53 maintains genome stability by arresting cells for damage repair or inducing cell apoptosis to eliminate the damaged cells in stress response. Several nucleolar proteins stabilize p53 by interfering Mdm2–p53 interaction upon cellular stress, while other mechanisms by which nucleolar proteins activate p53 remain to be determined. Here, we identify NAT10 as a novel regulator for p53 activation. NAT10 acetylates p53 at K120 and stabilizes p53 by counteracting Mdm2 action. In addition, NAT10 promotes Mdm2 degradation with its intrinsic E3 ligase activity. After DNA damage, NAT10 translocates to nucleoplasm and activates p53‐mediated cell cycle control and apoptosis. Finally, NAT10 inhibits cell proliferation and expression of NAT10 decreases in human colorectal carcinomas. Thus, our data demonstrate that NAT10 plays a critical role in p53 activation via acetylating p53 and counteracting Mdm2 action, providing a novel pathway by which nucleolar protein activates p53 as a cellular stress sensor.

Effects of RNA interference-mediatedMCM7knockdown on biological behavior of human liver cancer SMMC-7721 cells

Effects of RNA interference-mediated<i>MCM7</i>knockdown on biological behavior of human liver cancer SMMC-7721 cells

All-trans retinoic acid and oxaliplatin synergistically inhibit the proliferation of human liver cancer SMMC-7721 cells

All-trans retinoic acid and oxaliplatin synergistically inhibit the proliferation of human liver cancer SMMC-7721 cells

Effects of siRNA-targeting BMP-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells and its mechanism

To observe the effects of small interfering RNA (siRNA)-targeting bone morphogenetic protein (BMP)-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells and its mechanism. Three siRNAs-targeting BMP-2 gene were synthesized. There were six groups including group I (non-transfected cells), group II (only liposome-transfected cells), group III (non-specific siRNA-transfected cells) and groups IV-VI (siRNA-A, siRNA-B and siRNA-C-targeting BMP-2 transfected cells, respectively). SMMC7721 cells were instantaneously transfected using lipofectamine method. The levels of mRNA and protein of BMP-2 in cells were determined with reverse transcription-PCR and western blotting. The abilities of migration and invasion of transfected cells were assessed using scratch test and in vitro invasion assay, respectively. The protein levels of p-ERK, p-JNK and p-p38 and the protein levels of MMP-2 and MMP-9 were evaluated with western blot 48 h after siRNA-B-targeting BMP-2 was transfected into liver cancer SMMC7721 cells. Expression of mRNA and protein of BMP-2 in groups IV-VI were significantly inhibited, especially in group V. Cell scratch width was significantly greater in group V than in group I and III (P<0.01). In vitro invasion assay suggested that the number of invasion of cells was significantly lower in group V than in group I and III (P<0.05). Western blot indicated that the level of p-ERK was significantly decreased (P<0.05), the levels of p-JHK and p-p38 were not significantly changed and the levels of MMP-2 and MMP-9 were significantly downregulated (P<0.05). siRNA-targeting BMP-2 can markedly inhibited the expression of BMP-2 in liver cancer SMMC7721 cells, and decrease the abilities of migration and invasion of liver cancer cells, especially siRNA-B. The inhibitory effects of siRNA-B-targeting BMP-2 on the abilities of migration and invasion of human liver cancer SMMC7721 cells may be caused by the downregulation of MMP-2 and MMP-9 through MAPK/ERK pathway, whereas is not related to MAPK/JNK and MAPK/p38 pathway.

In vivo antitumor activity by 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone in a solid human carcinoma xenograft model

Previously we have shown that 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC), which is isolated from the buds of Cleistocalyx operculatus, significantly inhibits the growth of human liver cancer SMMC-7721 cells and is able to induce apoptosis of SMMC-7721 cells in vitro. Here we report the antitumor effects of DMC in vivo, using a solid human tumor xenograft mouse model using human liver cancer SMMC-7721 cells. The average tumor weights in the control group and in mice injected with 150 mg/kg DMC were 1.42+/-0.11 g and 0.59+/-0.12 g, respectively. Flow cytometric analysis of the tumor cell population demonstrated an aneuploid peak (representing 33.60+/-0.80% of the total in mice injected with 150 mg/kg DMC). To our knowledge, this is the first time that chalcone compounds have been applied to a human tumor xenograft model.

In vivo antitumor activity by 2?,4?-dihydroxy-6?-methoxy-3?,5?-dimethylchalcone in a solid human carcinoma xenograft model

Previously, we showed that 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalcone (DMC), isolated from the buds of Cleistocalyx operculatus, significantly inhibited the growth of human liver cancer SMMC-7721 cells and could induce SMMC-7721 cells apoptosis in vitro. Here, we report the antitumor effects of DMC in vivo, using a solid human tumor xenograft model with a human liver cancer SMMC-7721 cell line. Our results revealed that the average tumor weight in a control group and a 150-mg/kg DMC injection group was 1.42+/-0.11 g and 0.59+/-0.12 g, respectively. Flow cytometric analysis of the tumor cell population demonstrated the existence of an aneuploid peak (representing 33.60+/-0.80% of the total in the 150-mg/kg DMC injection group). To our knowledge, this is the first time that chalcone compounds were applied to a human tumor xenograft model.

Oral adeno‐associated virus‐sTRAIL gene therapy suppresses human hepatocellular carcinoma growth in mice†

The extracellular domain of the tumor necrosis factor-related apoptosis-inducing ligand (sTRAIL) may function as a soluble cytokine to selectively kill various cancer cells without toxicity to most normal cells. We constructed a series of recombinant adeno-associated virus (AAV) vectors expressing the extracellular domain of human TRAIL fused with signal peptides of human insulin, interferon, human growth hormone, and serum albumin and designated them as AAV-ISN-T, AAV-IFN-T, AAV-HGH-T, and AAV-Alb-T, respectively. Transduction of human SMMC-7721 liver cancer cells with AAV-ISN-T led to higher levels of TRAIL(95-281) protein expression in the cell culture media and produced more apoptosis of the cells in vitro than those with AAV-IFN-T, AAV-HGH-T, and AAV-Alb-T. The therapeutic potential of AAV-ISN-T was then evaluated in a transplanted mouse model established by injection of human liver cancer SMMC-7721 cells subcutaneously. Subsequent oral or intraperitoneal administration of AAV-ISN-T resulted in a rapid, high level and long time expression of soluble TRAIL in the sera and livers of the animals, as well as effective suppression of tumor growth, with no toxicity to normal hepatocytes. These data strongly suggest that it is possible to increase soluble TRAIL expression to make full use of tumoricidal activity of TRAIL as a therapeutic strategy. In conclusion, we provide evidence that oral administration of AAV-TRAIL might be an important alternative route with practical significance for cancer gene therapy.