Retraction: Down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in hepatocellular carcinoma cells via inactivation of the PI3K/Akt pathway.

[This retracts the article DOI: 10.1039/C8RA06860G.].

MiR-6991-3p is identified as a novel suppressor in the expansion and activation of myeloid-derived suppressor cells in hepatoma-bearing mice.

OBJECTIVE: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous group of cells derived from bone marrow, which has a significant ability in inhibition of immune cell response. In this study, the role of miR-6991-3p in regulating function of MDSCs was investigated. METHODS: MDSCs were isolated from different tissues of the control and hepatoma-bearing mice, and then expression of miR-6991-3p was detected with qPCR. Then, the miR-6991-3p mimic and inhibitor were respectively transfected into MDSCs, and behaviors of MDSCs were evaluated, including expansion, apoptosis, and production of inflammatory factors. Furthermore, we explored the underlying mechanism from which miR-6991-3p regulated MDSC functions. RESULTS: Expression miR-6991-3p was markedly decreased in the MDSCs derived from spleen and further decreased in the MDSCs derived from the tumor tissue. MiR-6991-3p mimic transfection suppressed expansion and promoted apoptosis of MDSCs, accompanied by a significant decrease in the production of IL-6 and GM-CSF that are identified as stimulators in MDSC expansion. In contrast, miR-6991-3p inhibitor transfection displayed the opposite effect. miR-6991-3p bound with and negatively regulated expression of LGALS9, a newly identified immune checkpoint gene and activator of STAT3, suppressing production of multiple factors that were customarily used to characterize the activation of MDSCs. MiR-6991-3p-accommodated MDSCs displayed less suppression on T cells, while miR-6991-3p inhibitor enhanced the suppression of MDSCs on T cells. CONCLUSION: MiR-6991-3p is identified as a novel suppressor in the expansion and activation of myeloid-derived suppressor cells, which may be regarded as a promising target for modulating the function of MDSCs.

Down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in hepatocellular carcinoma cells via inactivation of the PI3K/Akt pathway.

HOXB5, a member of the HOX gene family, is a developmental gene which encodes homeoproteins and is known to be a crucial player in development of enteric nervous systems. Recently, HOXB5 was reported to be associated with cancer progression. However, the specific effect of HOXB5 in hepatocellular carcinoma (HCC) remains unclear. In this study, we demonstrated the important role of HOXB5 in HCC. We showed that HOXB5 was up-regulated in HCC tissues and cell lines. Furthermore, down-regulation of HOXB5 inhibited TGF-ß-induced HCC cell migration and invasion in vitro and suppressed tumor metastasis in vivo. We also found that the PI3K/Akt pathway partly accounted for the mechanisms underlying the inhibitory effect of HOXB5 down-regulation on TGF-ß-induced HCC progression. Taken together, these findings demonstrated that down-regulation of HOXB5 inhibits TGF-ß-induced migration and invasion in HCC cells via inactivation of the PI3K/Akt pathway. Thus, HOXB5 may be a novel therapeutic target for HCC treatment.

The efficacy and safety of different pharmacological interventions for patients with advanced biliary tract cancer: A network meta-analysis.

Biliary tract cancer (BTC) is the second common cancer in liver cancer. Chemotherapy is the mainstay of treatments for patients with advanced or metastatic disease, while fluorouracil (FU)-based and gemcitabine (GEM)-based treatments are most widely applied. This NMA aimed to figure out whether the addition of platinum (PLA) and target agents (TAR) can influence the efficacy and safety of standard chemotherapy. Network meta-analysis (NMA) was conducted based on the records from PubMed, Embase and Cochrane. Eligible data was extracted from available qualified trials and outcomes. Software R 3.2.3 and STATA 13.0 were used to conduct the Bayesian NMA, calculating odds ratios (ORs) and hazard ratios (HRs) with 95% credible interval (CrI) to evaluate different treatments.Almost all treatments were superior to best supportive care (BSC) and FU in terms of 1-OS, 2-OS and 1-PFS. GEM+PLA and GEM+PLA+TAR exhibited better efficacy than most treatments in 1-OS, 2-OS and 1-PFS, and yielded better results than BSC and GEM+FU in terms of 2-PFS. Most drug-containing treatments reported higher overall response rate (ORR) than BSC. GEM and GEM+FU were associated with a higher risk of neutropenia and thrombocytopenia compared to FU, FU+PLA and GEM+PLA. No statistical difference was detected in terms of nausea and vomiting.GEM+PLA and GEM+PLA+TAR were both efficacious and were associated with fewer adverse events. In conclusion, the addition of PLA can significantly improve the efficacy of FU and GEM-based treatments, and the addition of TAR to GEM+PLA can contribute to further improvement, but with a mild increase of adverse events.

MicroRNA-27b exerts an oncogenic function by targeting Fbxw7 in human hepatocellular carcinoma.

Aberrant expression of microRNAs (miRNAs) plays fundamental effect on the pathogenesis of hepatocellular carcinoma (HCC). MiR-27b was previously found to play important roles in human cancers. However, its expression status, clinical significance, and biological functions in HCC remain largely unclear. The expression status of miR-27b in HCC specimens and cells were determined with qRT-PCR. MTT, 5-bromodeoxyuridine (BrdU) proliferation assays, and flow cytometry analysis were carried out to assay proliferation, cell cycle, and apoptosis. A subcutaneous model was used to evaluated the HCC tumor growth in vivo. The putative target gene of miR-27b was disclosed by TargetScan and a luciferase reporter assay. The levels of miR-27b were overexpressed in HCC. Overexpression of miR-27b was correlated with adverse prognostic features and reduced survival rate. Inhibition of miR-27b in SMMC-7721 cells remarkably suppressed proliferative ability and cell-cycle progression while enhanced apoptosis. In contrast, miR-27b overexpression resulted in prominent increased proliferation and process of cell cycle and reduced apoptosis of Hep3B cells. In vivo studies showed that knockdown of miR-27b inhibited the in vivo growth of SMMC-7721 cells in mouse xenograft model. Furthermore, we confirmed that Fbxw7 was directly regulated by miR-27b and mediated the roles of miR-27b in HCC. We suggest that miR-27b serves as an oncogenic miRNA in HCC by modulating proliferation, cell-cycle progression, and apoptosis, and its oncogenic effect is mediated by its downstream target gene, Fbxw7.

Trimethoxy-benzaldehyde levofloxacin hydrazone inducing the growth arrest and apoptosis of human hepatocarcinoma cells.

BACKGROUND: In order to search for new structural modification strategies on fluoroquinolones, we have designed and synthesized a series of fluoroquinolone derivatives by linking various hydrazine compounds to the C-3 carboxyl group of levofloxacin and assessed their anticancer activities. Several novel levofloxacin derivatives displayed potent cytotoxicity against the tested cancer cell lines in vitro. In the present study, we investigated the effect of 1-Cyclopropyl-6-fluoro-4-oxo-7- piperazin-1, 4-dihydro- quinoline- 3-carboxylic acid benzo [1,3] dioxol-5- ylmethylene- hydrazide (QNT11) on the apoptosis of human hepatocarcinoma cells in vitro. METHODS: The inhibition effects of QNT11 on cell proliferation were examined by MTT assay. Cell apoptosis was determined by TUNEL and DNA agarose gel electrophoresis method. The topoisomerase ΙΙ activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Cell cycle progression was analyzed using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Mitochondrial membrane potential (△ψm) was measured by high content screening image system. The caspase-9, caspase-8, caspase-3, Bcl-2, Bax, CDK1, Cyclin B1and cytochrome c protein expressions were detected by Western blot analysis. RESULTS: QNT11 showed selective cytotoxicity against Hep3B, SMMC-7721, MCF-7 and HCT-8 cell lines with IC50 values of 2.21 µM, 2.38 µM, 3.17 µM and 2.79 µM, respectively. In contrast, QNT11 had weak cytotoxicity against mouse bone marrow mesenchymal stem cells (BMSCs) with IC50 value of 7.46 µM. Treatment of Hep3B cells with different concentrations of QNT11 increased the percentage of the apoptosis cells significantly, and agarose gel electrophoresis revealed the ladder DNA bands typical of apoptotic cells, with a decrease in the mitochondrial membrane potential. Compared to the control group, QNT11 could influence the DNA topoisomerase IIactivity and inhibit the religation of DNA strands, thus keeping the DNA in fragments. There was a significant increase of cytochrome c in the cytosol after 24 h of treatment with QNT11 and a decrease in the mitochondrial compartment. Observed changes in cell cycle distribution by QNT11 treated might be caused by insufficient preparation for G2/M transition. In addition, QNT11 increased the protein expression of Bax, caspase-9, caspase-8, caspase-3, as well as the cleaved activated forms of caspase-9, caspase-8 and caspase-3 significantly, whereas the expression of Bcl-2 decreased. CONCLUSIONS: Our results showed that QNT11 as a fluoroquinolone derivative exerted potent and selectively anticancer activity through the mechanism of eukaryotic topoisomerase II poisoning. The growth inhibition was in large part mediated via apoptosis-associated mitochondrial dysfunction and regulation of Bcl-2 signaling pathways.

[Effect of acupuncture on the p53 protein expression of mice with Alzheimer's disease].

OBJECTIVE: To observe the effect of acupuncture method for benefiting qi, regulating blood, supplementing the root, and cultivating the essence (BQRBSRCE) on the p53 protein expression of mice with Alzheimer's disease (AD). METHODS: SAMP8 mice were divided into the control group, the acupuncture group, and the non-acupoint group. The homologous SAMR1 control group was set up. Mice in the acupuncture group used acupuncture method for BQRBSRCE by needling at Tanzhong (RN17), Zhongwan (RN12), Qihai (RN6), and bilateral Xuehai (SP10), and bilateral Zusanli (ST36).Two fixed non-acupoints from bilateral ribs were needled in the non-acupoint group. The p53 protein expression in the cortex and hippocampus of mice was determined using Western blot. The pathological changes of neurons in the hippocampal CA1 region, the temporal lobe, and the occipital lobe were observed using HE staining. The expression of cortical p53-positive cells was detected by immunohistochemical assay. RESULTS: The p53 protein was highly expressed in the cortex of SAMP8, which was significantly down-regulated after acupuncture, showing statistical difference when compared with the SAMP8 control group (P < 0.05), but with no statistical difference when compared with the SAMR1 control group (P > 0.05). Needling at non-acupoints had no obvious effect on the p53 protein expression. There was no statistical difference in the p53 protein expression of the hippocampus (P > 0.05). CONCLUSION: Acupuncture method for BQRBSRCE could down-regulate the p53 protein expression in the brain of mice, improve the pathological state of brain cells, thus enhancing learning and memory capabilities of AD mice, improving their cognitive functions, with specificity of acupoints.

[Effects of electroacupuncture on the expression of nestin in endogenous neural stem cell in hippocampus in rats with focal cerebral ischemia-reperfusion].

OBJECTIVE: To explore the mechanism of electroacupuncture for treating focal cerebral ischemia-reperfusion injury. METHODS: Seventy-five Wistar rats were randomly divided into a control group, a model group and an electroacupuncture group, 25 cases in each group. The model of focal cerebral ischemia-reperfusion was established by inserting nylon thread into the internal carotid artery except the control group which was only separated of the carotid artery without occlusion. Electroacupuncture group was treated with electroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" and the other groups without electroacupuncture treatment. The number of nestin positive cells expression at 1st, 3rd, 7th, 14th and 21st days after focal cerebral ischemia reperfusion was observed by use of immunohistochemistry method. RESULTS: The number of nestin positive cells in electroacupuncture group at ischemia side was significantly more than that in the model group at 3rd, 7th, 14th and 21st days (P < 0.05, P < 0.01), and at contralateral ischemia side, the number of nestin positive cells in the electroacupuncture group was significantly more than that in the model group only at 7th day (P < 0.05). CONCLUSION: Electroacupuncture at "Baihui (GV 20)" and "Dazhui (GV 14)" in rats can increase the number of nestin positive cells in hippocampus after focal cerebral ischemia-reperfusion, which may be one of the important mechanisms of electroacupuncture in treating acute cerebral ischemic diseases.

Effects of acupuncturing Tsusanli (ST36) on expression of nitric oxide synthase in hypothalamus and adrenal gland in rats with cold stress ulcer.

AIM: To study the protective effect of acupuncturing Tsusanli (S(T)36) on cold stress ulcer, and the expression of nitric oxide synthase (NOS) in hypothalamus and adrenal gland. METHODS: Ulcer index in rats and RT-PCR were used to study the protective effect of acupuncture on cold stress ulcer, and the expression of NOS in hypothalamus and adrenal gland. Images were analyzed with semi-quantitative method. RESULTS: The ulcer index significantly decreased in rats with stress ulcer. Plasma cortisol concentration was up regulated during cold stress, which could be depressed by pre-acupuncture. The expression of NOS1 in hypothalamus increased after acupuncture. The increased expression of NOS2 was related with stress ulcer, which could be decreased by acupuncture. The expression of NOS3 in hypothalamus was similar to NOS2, but the effect of acupuncture was limited. The expression of NOS2 and NOS3 in adrenal gland increased after cold stress, only the expression of NOS1 could be repressed with acupuncture. There was no NOS2 expression in adrenal gland in rats with stress ulcer. CONCLUSION: The protective effect of acupuncturing Tsusanli (S(T)36) on the expression of NOS in hypothalamus and adrenal gland can be achieved.

[Effect of acupuncture on the expression of choline acetyltransferase mRNA in the brain of ovarietomized rats].

The purpose of the present study was to investigate the mechanism of the effect of estrogen on the production of acetylcholine in the brain and to study the regulatory role of acupuncture of Zusanli acupoint in acetylcholine production in the brain of ovariectomized rats. Experimental female Wistar rats were divided into three groups: intact group (INT), ovariectomized group (OVX), and ovariectomy and acupuncture group (OVX+AC). Radioimmunoassay (RIA) was used to measure the estrogen content in plasma. The mRNA expression of choline-acetyltransferase (ChAT) and glyceraldehyde phosphate dehydrogenase (GAPDH) in the brain of rats was measured by the RT-PCR technique and was tested by the method of agarose gel electrophoresis. The ChAT mRNA positive neurons in the hippocampus were observed by using in situ hybridization and the results were processed with a computerized image analysis system. The results are as follows. Compared with the control animals, the plasma estrogen level was significantly lowered in ovariectomized animals. However, the plasma estrogen level was higher in the OVX+AC group than that of the OVX group. The ChAT mRNA expression level of OVX+AC group was higher than that of the OVX group. The area and integral optical density of the ChAT mRNA positive neurons in the hippocampus increased more obviously in OVX+AC group than in the OVX group. The experimental results observed indicate that the expression of ChAT gene in the brain is possibly related to the estrogen level in the body. The expression of ChAT gene in the brain of the ovarietomized rat can be regulated by acupuncture of Zusanli acupoint and it may be one of the mechanisms that acupuncture increases acetylcholine content in the brain.