The aim of the present study was to investigate the antitumor effect and mechanism of action of hyperforin in hepatocellular carcinoma (HCC) SK-Hep1 cells in vitro. Cells were treated with different concentrations of hyperforin for different periods of time. Effects of hyperforin on cell viability, apoptosis signaling, and expression of anti-apoptotic and proliferative proteins [cellular FLICE-like inhibitory protein (c-FLIP), X-linked inhibitor of apoptosis protein (XIAP), myeloid cell leukemia 1(MCL1), and cyclin-D1] were investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, and western blotting. Hyperforin significantly inhibited cell viability and expression of anti-apoptotic and proliferative proteins. We also found that hyperforin significantly induced accumulation of cells in sub-G phase, loss of mitochondrial membrane potential, and increased levels of active caspase-3, and caspase-8. Taken together, our findings indicate that hyperforin triggers inhibition of tumor cell growth by inducing intrinsic and extrinsic apoptotic pathways in HCC SK-Hep1 cells.

The self-synthesis of tungsten oxide (W18O49) nanowires on sputter-deposited W films prepared under different O2/Ar flow rate ratios (OAFRRs) in the sputtering gas is reported. After thermally annealing at 700–850 °C in N2 ambient for 15 min, dense and well crystalline W18O49 (010) nanowires or nanobelts were obtained depending on the oxygen content in the sputtering gas. Experimental results show that the annealing temperature required for the full growth of W18O49 nanowires reduced when the OAFRR in the sputtering gas was increased. It is found that the oxygen absorbed in the surface region is responsible for the growth of nanowires. As the OAFRR was increased to (8 sccm)/(24 sccm), which resulted in a saturated oxygen content of about 55 at.% inside the W film, large-scale nanobelts or nanosheets of W18O49 were grown. The possible growth mechanism which governs the evolution from nanowires to nanobelts as the OAFRR was changed is also discussed.

Glioblastoma is the most common primary malignant tumor of the central nervous system, with an annual incidence of 5.26 per 100000 people. The clinical outcome of standard therapy and the survival rate remain poor; therefore, there is an unmet need for a new strategy to treat this lethal disease. Although amentoflavone was known to have anticancer potential in various types of cancers, its antiglioblastoma ability and mechanism remain unrecognized. We demonstrated that amentoflavone may suppress glioblastoma invasion and migration by transwell assay. Moreover, we established NF-[Formula: see text]B reporter gene system and used that for verifying NF-[Formula: see text]B inhibition efficacy of amentoflavone on in vitro and in vivo studies. Here, we indicated that amentoflavone not only diminished NF-[Formula: see text]B activation, but also reduced NF-[Formula: see text]B-mediated downstream oncogenes expression, such as MMP-2, MMP-9, XIAP, cyclinD1 and VEGF, which was elucidated by Western blot and immunohistochemistry (IHC). Tumor growth inhibition and NF-[Formula: see text]B reduction was found in the amentoflavone treatment group, which was revealed by the glioblastoma-bearing animal model. In this study, we also used ERK inhibitor and NF-[Formula: see text]B inhibitor (QNZ) to confirm whether the beneficial result of amentoflavone on glioblastoma was mainly regulated by blockage of ERK/NF-[Formula: see text]B signaling. In summary, ERK/NF-[Formula: see text]B signaling pathway has a role in the inhibition of tumor growth by amentoflavone in glioblastoma.

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Abstract. The present study aimed to evaluate the effects of amentof lavone on sorafenib-induced apoptosis in sorafenib-resistant hepatocellular carcinoma (HCC) cells. The sorafenib-resistant SK-Hep1 (SK-Hep1R) cell line was established for the present study. Initially, the differences in sorafenib-induced cytotoxicity and apoptosis between wild-type SK-Hep1 and SK-Hep1R cells were verified using the MTT assay and flow cytometry. The effects of amentoflavone on sorafenib-induced cytotoxicity and apoptosis were then investigated using MTT, flow cytometry, DNA gel electrophoresis and western blot analysis. The results demonstrated that cell viability of SK-Hep1R cells was increased compared with that of SK-Hep1 cells following treatment with different concentrations of sorafenib for 24 h. Apoptosis of SK-Hep1R cells was lower than that of SK-Hep1 cells following treatment with 20 µM sorafenib for 24 h. Amentoflavone alone did not inhibit cell viability but significantly triggered sorafenib-induced cytotoxicity and apoptosis in SK-Hep1R cells. Amentoflavone not only reversed sorafenib-induced anti-apoptotic protein levels but also enhanced sorafenib-induced pro-apoptotic protein expression in SK-Hep1R cells. In conclusion, amentoflavone may be used as a sorafenib sensitizer to enhance sorafenib-induced cytotoxicity and trigger sorafenib-induced apoptosis through extrinsic and intrinsic pathways in SK-Hep1R cells. IntroductionSorafenib, a multi-kinase inhibitor, has been approved by the US Food and Drug Administration to improve overall survival and time to progression of patients with advanced hepatocellular carcinoma (HCC) (1). Sorafenib induces apoptosis and inhibits angiogenesis in HCC through blockage of the rapidly accelerated fibrosarcoma/mitogen-activated protein kinase/extracellular signal-regulated kinase cascade, vascular endothelial growth factor and platelet-derived growth factor receptor tyrosine kinase signaling (2,3). Sorafenib has also been demonstrated to enhance the therapeutic efficacy of anticancer agents and radiotherapy via inhibition of nuclear factor-κB (NF-κB) or signal transducer and activator of transcription 3 (STAT3)-modulated resistance to anticancer treatments in HCC models in vitro and in vivo (4,5). However, long-term exposure to sorafenib for HCC cells induces sorafenib resistance and results in tumor progression (6,7). Therefore, development of sorafenib sensitizers, which reverse sorafenib resistance and results in sorafenib-inhibited tumor progression in sorafenib-resistant HCC cells, is important. Amentoflavone enhances sorafenib-induced apoptosis through extrinsic and intrinsic pathways in sorafenib-resistant hepatocellular carcinoma SK-Hep1 cells in vitro

BackgroundNuclear factor (NF)-κB signaling in cancer cells was reported to be involved in tumorigenesis.Phosphorylation and degradation of inhibitor of NF-κBα (IκBα) is a canonical pathway of NF-κB signaling. Herein, we report non-canonical activation of NF-κB signaling without phosphorylation of IκB but by directly binding by ubiquitin-conjugating enzyme E2S (UBE2S) for degradation in adenocarcinoma cells. MethodsTCGA and the Human Atlas Protein Database were used to analyze the survival rate and expression of UBE2S. PC9, H460, H441 and A549 cells were used in this study. PC9 and H460 cells were used for further analysis because of different protein levels of UBE2S. Speci c IKK inhibitors, PS1145 and SC514, were used to analyze the phosphorylation of IκBα. Western blotting experiment was used to analyze the protein levels PC9 and H460 cells. Wound-healing experiment was used to analyze the migrative ability of PC9 and H460 cells. Overexpression and knockdown of UBE2S in H460 and PC9 cells were used to analyze the downstream proteins levels. Immunoprecipitation, immuno uorescent staining, a glutathione S transferase (GST) pull-down assay, and an in vitro binding assay were used to analyze the interaction of UBE2S and IκBα. Luciferase assay was used to analyze the activation of NF-κB signaling regulated by UBE2S. Zebra sh xenograft model was used analyzed the metastasis of PC9 cells regulated by UBE2S. ResultsUBE2S in lung adenocarcinoma patients was negatively related to the survival rate. The protein levels of UBE2S and IκBα were shown opposite change in PC9 and H460 cells. PC9 cells showed higher UBE2S expression and migrative ability than H460 cells. Phosphorylation of IκBα was not changed by treatment with IKK speci c inhibitors, PS1145 and SC514, in PC9 and H460 cells. Overexpression and knockdown of UBE2S in H460 and PC9 cells showed the protein levels of IκBα were regulated. Immunoprecipitation, immuno uorescent staining, a glutathione S transferase (GST) pull-down assay, and an in vitro binding assay showed the direct binding of UBE2S with IκBα. Protein levels of nuclear p65 and luciferase assay showed the NF-κB signaling was regulated UBE2S. EMT markers and migrative ability of cancer cells were also regulated by UBE2S. Zebra sh xenograft tumor model showed the reduction of migrative PC9 cells by knockdown of UBE2S. ConclusionHigher UBE2S expressed in lung adenocarcinomas could bind with IκBα for activation of NF-κB signaling to promote metastasis of cancer cells. UBE2S might be a potential therapeutic target for lung adenocarcinomas.

Background The aim of the study was investigate the impact of body-mass factors (BMF) on setup displacement during pelvic radiotherapy in patients with lower abdominal cancers. Patients and methods The clinical data of a training cohort composed of 60 patients with gynecological, rectal, or prostate cancer were analyzed. The daily alignment data from image-guided radiotherapy (IGRT) were retrieved. Setup errors for were assessed by systematic error (SE) and random error (RE) through the superior-inferior (SI), anterior-posterior (AP), and medial-lateral (ML) directions. Several BMFs and patient-related parameters were analyzed with binary logistic regression and receiver-operating characteristic curves. A scoring system was proposed to identify those with greater setup displacement during daily treatment. The results were validated by another cohort. Results A large hip lateral diameter correlated with a greater SI-SE and AP-SE, whereas a large umbilical AP diameter correlated with a greater ML-SE and ML-RE. A higher SI-RE was associated with a large hip circumference. The positive predictors for setup uncertainty were chosen to dichotomize patients into groups at high risk and low risk for setup displacement. Based on the scoring system, the adequate treatment margins for the SI direction in the high-and low-risk groups were 5.4 mm and 3.8 mm, whereas those for the ML direction were 8.2 mm and 4.2 mm, respectively. The validated cohort showed a similar trend. Conclusions Large BMFs including hip lateral diameter, hip circumference, and umbilical AP diameter are associated with greater setup uncertainty. Based on the scores, IGRT or required treatment margins can be adapted for patients with high risk features.

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