Description: The paper aimed to explore the specific function and mechanism of miR-144-3p in glioblastoma (GBM) cells with different PTEN (Phosphatase and Tensin homolog) phenotypes. We demonstrated that the miR-144-3p level was significantly down-regulated in glioma compared with the non-neoplastic brain tissues, and decreased with ascending grades. The loss of miR-144-3p effectively predicted the decreased overall survival in glioma patients. Interestingly, the expression of MET was up-regulated and inversely associated with miR-144-3p level in glioma tissues. Next, we certified that miR-144-3p specifically bound to MET 3′ UTR and inhibited its expression. miR-144-3p potently repressed GBM cell proliferation and invasion via suppressing MET in vitro and in vivo. In addition, our results showed no difference in malignancy inhibition induced by miR-144-3p in GBM cells with different PTEN phenotypes. miR-144-3p inhibited several survival signaling pathways by targeting MET independent of PTEN status in GBM cells. Over-expression of miR-144-3p inhibited survival capability and increased apoptosis, resulting in enhancement of radiation and temozolomide (TMZ) sensitivity. Our data provide new insights into the potential application of miR-144-3p in GBM therapy by targeting MET and then inhibiting downstream signaling. This article is protected by copyright. All rights reserved.

Description: Selection of dose for cancer patients treated with radiation therapy (RT) must balance the increased efficacy with the increased toxicity associated with higher dose. Historically, a single dose has been selected for a population of patients (e.g., all stage III non-small cell lung cancer). However, the availability of new biologic markers for toxicity and efficacy allows the possibility of selecting a more personalized dose. We consider the use of statistical models for toxicity and efficacy as a function of RT dose and biomarkers to select an optimal dose for an individual patient, defined as the dose that maximizes the probability of efficacy minus the sum of weighted toxicity probabilities. This function can be shown to be equal to the expected value of the utility derived from a particular family of bivariate outcome utility matrices. We show that if dose is linearly related to the probability of toxicity and efficacy, then any marker that only acts additively with dose cannot improve efficacy, without also increasing toxicity. Using a dataset of lung cancer patients treated with RT, we illustrate this approach and compare it to non-marker-based dose selection. Because typical metrics used in evaluating new markers (e.g., area under the ROC curve) do not directly address the ability of a marker to improve efficacy at a fixed probability of toxicity, we utilize a simulation study to assess the effects of marker-based dose selection on toxicity and efficacy outcomes. Copyright © 2014 John Wiley & Sons, Ltd.

Description: Invariant natural killer T (iNKT) cells are a major subset of lymphocytes found in the liver. These cells mediate various functions, including hepatic injury, fibrogenesis, and carcinogenesis. However, the function of iNKT cells in liver regeneration remains unclear. In the present study, partial hepatectomy (PHx) was used to study liver regeneration. α-GalCer, a specific ligand for iNKT cells, was used to induce iNKT cell activation. After PHx, two strains of iNKT cell-deficient mice, CD1d -/- and Jα281 -/- mice, showed normal liver regeneration. Injection of α-GalCer before or after PHx, which rapidly stimulated IFN-γ and IL-4 production by iNKT cells, markedly inhibited liver regeneration. In vitro treatment with IFN-γ inhibited hepatocyte proliferation. In agreement with this in vitro finding, genetic disruption of IFN-γ or its downstream signaling molecule signal transducer and activator of transcription (STAT)1 significantly abolished the α-GalCer-mediated inhibition of liver regeneration. In vitro exposure to IL-4 did not affect hepatocyte proliferation, but surprisingly, genetic ablation of IL-4 or its downstream signaling molecule STAT6 partially eliminated the inhibitory effect of α-GalCer on liver regeneration. Further studies revealed that IL-4 contributed to α-GalCer-induced iNKT cell expansion and IFN-γ production, and thereby inhibiting liver regeneration. Conclusions : iNKT cells play a minor role in controlling liver regeneration after PHx under healthy conditions. Activation of iNKT cells by α-GalCer induces the production of IFN-γ, which directly inhibits liver regeneration, and IL-4, which indirectly attenuates liver regeneration by stimulating iNKT cell expansion and IFN-γ production. (H epatology 2014;)

Description: Berberine, an isoquinoline alkaloid isolated from several traditional Chinese herbal medicines, has been shown to suppress growth and induce apoptosis in some tumor cell lines. However, berberine has also been reported to attenuate H 2 O 2 -induced oxidative injury and apoptosis. The basis for these ambiguous effects of berberine—triggering or preventing apoptosis—has not been well characterized to date. In the current investigation, we examined whether berberine exerts cytotoxic effects on mouse embryos at the blastocyst stage and affects subsequent embryonic development in vitro and in vivo . Treatment of blastocysts with berberine (2.5-10 μM) induced a significant increase in apoptosis and a corresponding decrease in trophectoderm cell number. Moreover, the implantation success rate of blastocysts pretreated with berberine was lower than that of their control counterparts. Pretreatment with berberine was also associated with increased resorption of postimplantation embryos and decreased fetal weight. In an animal model, intravenous injection of berberine (2, 4, or 6 mg/kg body weight/d) for 4 days resulted in apoptosis of blastocyst cells and early embryonic developmental injury. Berberine-induced injury of mouse blastocysts appeared to be attributable to oxidative stress-triggered intrinsic apoptotic signaling processes that impaired preimplantation and postimplantation embryonic development. Taken together, our results clearly demonstrate that berberine induces apoptosis and retards early preimplantation and postimplantation development of mouse embryos, both in vitro and in vivo .

Description: Cantharidin analogs exhibit anticancer activities, including apoptosis. However, the molecular mechanisms underlying the effects of cantharidic acid (CA), a cantharidin analog, on apoptosis in hepatocellular carcinoma (HCC) cells are unclear. Thus, in this study, we evaluated the anticancer activities of CA by investigating its ability to trigger apoptosis in SK-Hep-1 cells. Our data demonstrated that CA effectively inhibited the proliferation of SK-Hep-1 cells in a dose-dependent manner. Furthermore, CA effectively triggered cell cycle arrest and induced apoptosis, as determined by flow cytometric analysis. Western blotting revealed that CA significantly activated proapoptotic signaling including caspase-3, −8, and −9 in SK-Hep-1 cells. Moreover, treatment of SK-Hep-1 cells with CA induced the activation of ERK, p38, and c-Jun N-terminal kinase. Moreover, the inhibition of p38 by specific inhibitors abolished CA-induced cell apoptosis. In conclusion, our results indicated that CA induces apoptosis in SK-Hep-1 cells through a p38-mediated apoptotic pathway and could be a new HCC therapeutic agent.

Description: Several strategies to improve the efficacy of radiation therapy against hepatocellular carcinoma (HCC) have been investigated. One approach was to develop radiosensitizing compounds. Because histone deacetylase 4 (HDAC4) is highly expressed in liver cancer and known to regulate oncogenesis through chromatin structure remodeling and controlling protein access to DNA, we postulated that HDAC4 inhibition might enhance radiation's effect on HCC cells. HCC cell lines (Huh7 and PLC5) and an ectopic xenograft were pretreated with HDAC inhibitor or shRNA to knock down expression of HDAC4, and then irradiated (2.5-10 Gy). We evaluated cell survival by a clonogenic assay; apoptosis by annexin-V immunofluorescence; γH2AX, Rad51, and HDAC4 by immunofluorescence staining; HDAC4, Rad51, and Ubc9 in HCC cell nuclei by cell fractionation and confocal microscopy; physical interaction between HDAC4/Rad51/Ubc9 by immunoprecipitation; the downstream targets of HDAC4 knockdown by immunoblotting. Both HDAC4 knockdown and HDAC inhibitor enhanced radiation-induced cell death, and reduced homologous recombination repair of DNA double-strand breaks and Akt activation, leading to increased apoptosis. HDAC4 knockdown with or without an HDAC inhibitor significantly delayed tumor growth in a radiation-treated xenograft model. Radiation stimulated nuclear translocation of Rad51 in an HDAC4-dependent manner and the binding of Ubc9 directly to HDAC4, which led to Ubc9 acetylation. Moreover, these effects were accompanied by HDAC4/Ubc9/Rad51 complex dissociation through inhibiting nuclear translocation. Conclusions : HDAC4 signaling blockade enhances radiation-induced lethality in HCC cells and xenografts. These findings raise the possibility that HDAC4/Ubc9/Rad51 complex in DNA repair may be a target for radiosensitization of HCC. This article is protected by copyright. All rights reserved.

Description: Purpose To explore the morphological and functional characteristics of prostatic arterial embolization (PAE) in a canine model of benign prostatic hyperplasia (BPH) with 3T multiparametric magnetic resonance imaging (mp-MRI) and whole-mount step-section pathology correlation. Materials and Methods Eight adult male beagle dogs with hormone-induced BPH underwent 3T mp-MRI before and 1, 3, and 6 months after PAE, with subsequent whole-mount step-section pathologic assessment. Images were acquired using T 1 -weighted images ( T 1 WI), T 2 WI, 3D-SPACE, diffusion-weighted imaging (DWI), susceptibility-weighted imaging (SWI), T 2 -mapping, and dynamic contrast-enhanced (DCE) sequences. Variance analysis was performed to assess statistical differences in prostatic volume (PV), apparent diffusion coefficient (ADC), and T 2 values. Pearson correlation analysis was performed to correlate ADC, T 2 , and PV. Results The PV decreased from baseline to 1, 3, and 6 months after PAE from (25.88 ± 7.09) cm 3 to (6.48 ± 2.08) cm 3 , (6.48 ± 3.39) cm 3 , (6.20 ± 2.88) cm 3 . The ADC values sequentially decreased from baseline to 1, 3, and 6 months after PAE from (1497.06 ± 222.72) × 10 −6 mm 2 /s to (1056.00 ± 189.46) × 10 −6 mm 2 /s, (950.48 ± 77.85) × 10 −6 mm 2 /s, (980.98 ± 107.78) × 10 −6 mm 2 /s. The T 2 values decreased from baseline to 1, 3, and 6 months after PAE were (83.74 ± 5.29) msec, (68.72 ± 5.66) msec, (53.96 ± 15.04) msec, (49.81 ± 13.34) msec, respectively. ADC and T 2 values were positively correlated with PV ( r  = 0.823 and 0.744, respectively). Microhemorrhages and hemosiderin were found on SWI after PAE. Conclusion 3T mp-MRI may facilitate noninvasive assessment of morphological and functional changes of BPH after PAE. Level of Evidence: 1 J. Magn. Reson. Imaging 2017

Description: The p53 protein plays a critical role in suppression of tumour growth; its regulation is not fully understood. Leukaemia/lymphoma-related factor (LRF) promotes tumour cell growth. This study tests a hypothesis that LRF inhibits p53 expression in colon cancer cells. In this study, human colon cancer cell lines, LIM1215 and HCT116 cells, were used. The expression of LRF and p53 in the cells was analysed by quantitative reverse transcription polymerase chain reaction and Western blotting. We observed that the expression of protease-activated receptor 2 (PAR2) was detected in both LIM1215 and HCT116 human colon cancer cells. Activation of PAR2 increased the expression of LRF and inhibited the p53 expression in the cancer cells. We also detected a complex of LRF and DAP5, one of the p53 gene transcription factors. The interaction of LRF and DAP5 resulted in the repression of p53 expression in the colon cancer cells. In conclusion, PAR2 activation increases the expression of LRF in colon cancer cells, which interacts with DAP5 to repress the p53 expression. Leukaemia/lymphoma-related factor may be a novel target in the treatment of colon cancer.

Description: Background: Bone morphogenetic protein receptor II (BMPR-II) plays an important role in tumor's invasion and proliferation. In this study, we observed the effects of small interfering RNA (siRNA) targeting bone morphogenetic protein receptor II (BMPR-II) on the biological activities of human liver cells and explore its mechanism. Methods: The molecular sequences of three siRNA targeting BMPR-IIwere designed and synthesized. In this study, there were 6 groups including group I (normal control), group II (blank control), group III (negative control) and group IV-VI (BMPR-II-siRNA-a, siRNA-b and siRNA-c-transfected cells, respectively). The levels of mRNA and protein of BMPR-II were determined to select the best sequence for BMPR-II silence. After liver cancer cells were transfected with the best sequence, proliferation and invasion of transfected cells were assessed, and apoptosis and cell cycle were detected. The expressions of mitogen-activated protein kinases (MAPKs) signal pathway-related proteins were observed after BMPR-II silence and BMPR-II silence combined with inhibiting MAPKs signal pathway, respectively. Results: RT-PCR and Western blot indicated that BMPR-II expression was the highest in HepG2 among the three liver cancer lines (P 〈 0.01) and the lowest in group IV among the six groups (P 〈 0.01).MTT assay and transwell assay revealed that the numbers of cell growth and cell transmembrane were significantly lower in group IV than in control groups 48 h after cells were transfected (P 〈 0.05).Flow cytometer showed that apoptosis was the highest and cells were significantly blocked in S phase 48 h after cells were transfected in group IV (P 〈 0.01).Western blot indicated that the protein levels of p-P38(P 〈 0.01) and vascular endothelial growth factor-C (VEGF-C) (P 〈 0.01) were significantly decreased after BMPR-II silence. The protein level of VEGF-C was significantly decreased in PD98059 + siRNA-BMPR-II-a and SB203580 + siRNA-BMPR-II-a groups (P 〈 0.01), especially in SB203580 + siRNA-BMPR-II-a group (P 〈 0.01). Conclusions: siRNA targeting BMPR-IIcan markedly inhibit HepG2 proliferation and invasion, promote apoptosis and block HepG2 in S phase. Its mechanism may be that BMPR-II silence down-regulates VEGF-C expression through MAPK/P38 and MAPK/ERK1/2 pathways, especially MAPK/P38. This study provides a new targeted therapy for liver cancer.

Description: Temozolomide (TMZ) has been widely used in the treatment of glioblastoma (GBM), although inherent or acquired resistance restricts the application. The present study was aimed to evaluate the efficacy of sulforaphane (SFN) to TMZ-induced apoptosis in GBM cells and the potential mechanism. Biochemical assays and subcutaneous tumor establishment were used to characterize the function of SFN in TMZ-induced apoptosis. Our results revealed that β-catenin and miR-21 were concordantly expressed in GBM cell lines, and SFN significantly reduced miR-21 expression through inhibiting the Wnt/β-catenin/TCF4 pathway. Furthermore, down-regulation of miR-21 enhanced the pro-apoptotic efficacy of TMZ in GBM cells. Finally, we observed that SFN strengthened TMZ-mediated apoptosis in a miR-21-dependent manner. In conclusion, SFN effectively enhances TMZ-induced apoptosis by inhibiting miR-21 via Wnt/β-catenin signaling in GBM cells. These findings support the use of SFN for potential therapeutic approach to overcome TMZ resistance in GBM treatment. This article is protected by copyright. All rights reserved.