hPCL3s Promotes Hepatocellular Carcinoma Metastasis by Activating ß-Catenin Signaling.

Two isoforms of human Polycomb-like protein 3 (hPCL3) have been reported as components of the nuclear Polycomb repressive complex 2 (PRC2), with the short isoform (hPCL3s) showing a dominant cytoplasmic localization. The function of cytoplasmic hPCL3s has, however, not been addressed. In this study, we report that hPCL3s is upregulated in clinical hepatocellular carcinoma (HCC) samples and its expression correlated with HCC clinical features. hPCL3s positively regulated the migration, invasion, and metastasis of HCC cells. hPCL3s interacted with components of the cytoplasmic ß-catenin destruction complex, inhibited ß-catenin degradation, and activated ß-catenin/T-cell factor signaling. Downstream of the ß-catenin cascade, IL6 mediated the motility-promoting functions of hPCL3s. Forced expression of hPCL3s in the liver of a HCC mouse model promoted tumorigenesis and metastasis. Taken together, these data show that hPCL3s promotes the metastasis of HCC by activating the ß-catenin/IL6 pathway.Significance: hPCL3s has an oncogenic role in hepatocellular carcinoma by activating the ß-catenin/IL6 signaling axis to promote metastasis. Cancer Res; 78(10); 2536-49. ©2018 AACR.

Cilia loss sensitizes cells to transformation by activating the mevalonate pathway.

Although cilia loss and cell transformation are frequently observed in the early stage of tumorigenesis, the roles of cilia in cell transformation are unknown. In this study, disrupted ciliogenesis was observed in cancer cells and pancreatic cancer tissues, which facilitated oncogene-induced transformation of normal pancreatic cells (HPDE6C7) and NIH3T3 cells through activating the mevalonate (MVA) pathway. Disruption of ciliogenesis up-regulated MVA enzymes through ß catenin-T cell factor (TCF) signaling, which synchronized with sterol regulatory element binding transcription factor 2 (SREBP2), and the regulation of MVA by ß-catenin-TCF signaling was recapitulated in a mouse model of pancreatic ductal adenocarcinoma (PDAC) and human PDAC samples. Moreover, disruption of ciliogenesis by depleting Tg737 dramatically promoted tumorigenesis in the PDAC mouse model, driven by KrasG12D , which was inhibited by statin, an inhibitor of the MVA pathway. Collectively, this study emphasizes the crucial roles of cilia in governing the early steps of the transformation by activating the MVA pathway, suggesting that statin has therapeutic potential for pancreatic cancer treatment.

[Effects of Effective Fractions of Panax Notoginseng on the Expression of TF and TFPI-2 in Rat En- dometrium with Whole Cycle Exogenous Estrogen Intervention].

Objective To observe the effects of notoginsenoside, Panax notoginseng flavonoids (PNF) , Panax notoginseng acid (PNA) , and their mixtures on the expressions of tissue factor (TF) and tissue factor pathway inhibitor 2 (TFPI-2) in rat endometriaum with whole cycle exogenous estrogen inter- vention. Methods Totally 160 female impuberty rats were randomly divided into the blank group (n =40) and the model group (n =120). Rats in the blank group were administered with normal saline by gastroga- vage for 3 weeks, while those in the model group were administered with estradiol valerate suspension. The two interventions lasted for 3 consecutive weeks. After 3 weeks of intervention, rats with asexual cycle were randomly divided into six groups, i.e., the model group, the continuous estrogen group, the No- toginsenoside group, the PNF group, the PNA group, the mixture group. Rats in the model group, the continuous estrogen group, the Notoginsenoside group, the PNF group, the PNA group, the mixture group were respectively administered with normal saline, estrogen, notoginsenoside, PNF, PNA, and mixture of effective Panax notoginseng fractions by gastrogavage for 2 successive weeks. Expression levels of TF, TFPI-2, TF mRNA, and TFPI-2 mRNA in the endometrium were detected 2 weeks later. Re-sults Compared with the blank group, positive expressions of TF and TF mRNA increased, and positive expressions of TFPI-2 and TFPI-2 mRNA decreased in the model group (P <0. 05, P <0. 01). Compared with the model group, the expressions of TF and TF mRNA significantly increased, the expressions of TF- PI-2 and TFPI-2 mRNA significantly decreased in the estrogen group at the end of the 5th week (P < 0. 01). Compared with the model group and the estrogen group, positive expressions of TF and TF mRNA significantly decreased, positive expressions of TFPI-2 and TFPI-2 mRNA significantly increased in the Notoginsenoside group, the PNF group, the PNA group, the mixture group (P <0. 05, P <0. 01). Com- pared with the Notoginsenoside group, positive expressions of TFPI-2 and TFPI-2 mRNA significantly in- creased in the mixture group (P <0. 05). Compared with the PNF group, the expressions of TF and TF mRNA significantly decreased in the Notoginsenoside group and the mixture group (P <0. 01) ; positive expressions of TFPI-2 increased in the mixture group (P <0. 05, P <0. 01). Conclusions The effective fractions of Panax notoginseng could decrease the-expression of TF and increase the expression of TFPI- 2 in rat endometrium with whole cycle exogenous estrogen intervention. They activated blood circulation and arrested bleeding possibly through inhibiting TF, blocking activation of coagulation system, and re- ducing inflammatory response. Meanwhile, it also could strengthen endometrial extracellular matrix, maintain the endometrial stability, thereby reducing endometrial disintegration and bleeding, and being beneficial for endometrium repairing and remodeling.

[Up-regulation of ErbB3-binding protein 1 inhibits the growth of esophageal carcinoma cells].

The objective of this study was to investigate the role of ErbB3-binding protein 1 (Ebp1) in the growth of esophageal squamous cell carcinoma (ESCC) cells and the underlying mechanism. Eca109 and KYSE150 cells were transfected with lentiviral vector carrying Ebp1 gene. The mRNA levels of Ebp1 in esophageal cancer tissues and paired adjacent normal tissues were examined by real-time PCR. The growth and viability of esophageal carcinoma cells were assessed using MTT and crystal violet assays, respectively. Clone-forming abilities of Eca109 and KYSE150 cells were analyzed by soft agar assay. Apoptotic rates of esophageal carcinoma cells were detected by flow cytometry, and expression levels of the proteins involved in apoptosis were assessed by Western blot. Tumorigenicity of Eca109 cells were detected by nude mouse transplantation tumor experiment. The results indicated that the mRNA levels of Ebp1 in esophageal cancer tissues was down-regulated compared with paired adjacent normal tissues. The growth and viability of Eca109 and KYSE150 cells were all suppressed by Ebp1 overexpression. Besides, Ebp1 overexpression induced apoptosis, increased Rb and P53 expressions, and decreased CyclinD1 expression in Eca109 and KYSE150 cells. In addition, Ebp1 overexpression inhibited the tumorigenesis of Eca109 cells in vivo. These results suggest that Ebp1 may suppress the growth of esophageal carcinoma cells in vitro and in vivo by inducing apoptosis.

Furocoumarin derivatives from radix Angelicae dahuricae and their effects on RXRα transcriptional regulation.

A novel furocoumarin derivative named oxyalloimperatorin (1), together with seventeen furocoumarins 2-18 were isolated from the radix of Angelica dahurica. The chemical structure of new metabolite was characterized by analysis of IR, NMR, and HR-ESI-MS spectroscopic data. Among the isolated compounds, 13, 16, and 18 (each at 20 µM) could significantly promote the gene transcriptional function of nuclear receptor RXRα. While 7-9, 13, 14, and the new structure 1 (each at 20 µM) showed significant reduction in RXRα gene transcriptional activities induced by 9-cis-retinoid acid. The findings indicated that these furocoumarin skeleton derivatives might hold beneficial effects on many intractable diseases, such as cancer and metabolic diseases, due to their potential activities on regulating the transcriptional activation function of RXRα.

Traditional Chinese medicine Kang Xian Fu Fang I is effective for prophylaxis and treatment of alcoholic liver disease in rats.

BACKGROUND: Reversal of liver fibrosis is one of the key steps in the prevention and treatment of alcoholic liver disease (ALD), but the mechanism is unknown. This study was to investigate the effects of the Chinese medicine Kang Xian Fu Fang I (KXI) on prophylaxis and treatment of ALD in rats and its possible mechanism of action. METHODS: Eighty male Wistar rats were randomly divided into four groups: normal control; ALD model; treatment of ALD with KXI; and prophylaxis of ALD by KXI. At the end of 4, 8, 12 and 16 weeks, five rats from each group were anesthetized and their livers were removed for pathological studies using hematoxylin-eosin and Masson stain, immunohistochemical studies, and flow cytometry for matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). Blood samples were taken for hyaluronic acid (HA) assay. RESULTS: Serum HA level and liver collagen content were lower in the groups given KXI for prophylaxis and treatment than in ALD model group (P<0.05). The levels of MMP-2 and MMP-9 were also decreased in the prophylaxis and treatment groups (P<0.05). Immunohistochemistry showed immunoreactive MMP-2 in endothelial cells of the hepatic artery and portal vein, sinusoidal endothelial cells, and sinusoidal cells. Immunoreactive MMP-9 occurred in the hepatic cells around the veins and sinusoidal cells. CONCLUSIONS: KXI can effectively inhibit or reverse the course of ALD. This may be attributable to its capacity to inhibit the expression of MMP-2 and MMP-9.

Involvement of IFN regulatory factor (IRF)-1 and IRF-2 in the formation and progression of human esophageal cancers.

IFN regulatory factor (IRF)-1 and IRF-2 are generally regarded as a tumor suppressor and an oncoprotein, respectively. However, little is known about their expression and function in esophageal squamous cell carcinomas (ESCC). In our present work, IRF-1 expression was decreased and IRF-2 expression was increased in ESCCs compared with matched normal esophageal tissues. Moreover, statistical data indicated that IRF-2 expression was tightly correlated with progression of ESCCs. As expected, overexpression of either IRF-1 or IRF-2 in an ESCC cell line resulted in either suppression or enhancement of cell growth, respectively. Also, proliferation- and apoptosis-related molecules (p21(WAF1/CIP1), cyclin-D1, Bcl-2, and histone H4) were regulated by IRF-1 and IRF-2. Additionally, high levels of IRF-2 blocked the function of IRF-1 by preventing the latter from translocating into the nucleus; in contrast, knock down of IRF-2 by small interfering RNA permitted nuclear localization and activity of IRF-1. In vivo assay using nude mice indicated that the tumorigenicity of ESCC cells was enhanced with IRF-2 overexpression but dramatically attenuated after forced expression of IRF-1. In conclusion, IRF-1 and IRF-2 are able to regulate tumorigenicity of ESCC cells as antioncoprotein and oncoprotein, respectively. Relative amounts of IRF-1 to IRF-2 are functionally very important for the development and progression of ESCCs, and reduction of the ratio of IRF-1/IRF-2 may lead to the enhancement of tumorigenicity of ESCC cells. Therefore, levels of IRF-1 and IRF-2 are useful indicators in diagnosis and prognosis for ESCCs, and these molecules are potential drug targets for ESCC therapy.

Ephrin-A1 is a negative regulator in glioma through down-regulation of EphA2 and FAK.

Eph receptors, the largest receptor tyrosine kinases, and their ephrin ligands play important roles in axon guidance and cell migration during development of the nervous system. Recently, these molecules are also found involved in tumorigenesis of different kinds of cancers. In this study, we demonstrated that expression of ephrin-A1 was dramatically down-regulated in glioma cell lines and in primary gliomas compared to the matched normal tissues. Forced expression of ephrin-A1 attenuated cell migration, cell proliferation, and adhesion-independent growth in human glioma U251 cells. EphA2, a receptor for ephrin-A1 and an oncoprotein, was greatly decreased in ephrin-A1-transfected glioma cells. Overexpression of ephrin-A1 stimulated activation of EphA2 by phosphorylation and led to its degradation. Furthermore, focal adhesion kinase (FAK), a known downstream molecule of EphA2, was also down-regulated in the ephrin-A1 transfected cells. These results suggested that ephrin-A1 serves as a critical negative regulator in the tumorigenesis of gliomas by down-regulating EphA2 and FAK, which may provide potential valuable targets for therapeutic intervention.

Discovery of stage-related proteins in esophageal squamous cell carcinoma using proteomic analysis.

Esophageal squamous cell carcinoma (ESCC) is the major subtype of esophageal cancers in China, and characterized with high morbidity and mortality. So far, the diagnosis of ESCC is mainly dependent on the alterations in esophageal histology, but most cases of ESCC with low stage do not display visible histological abnormalities. Therefore, a deep understanding of the mechanism of ESCC progression and seeking stage-specific molecules might improve the diagnosis and therapy for ESCC. In this study, we used proteomics to analyze ESCC tissues with classification by TNM stage, and determined the proteomic features correlated with ESCC progression (from stages I to III). Proteins that exhibited significantly different expression patterns between ESCC and corresponding normal esophageal tissues were identified using MS. The identified proteins with differentiated expression mainly fell into three protein categories (i.e. cytoskeleton system-associated proteins, metabolism enzymes, and heat shock proteins). In addition, real-time PCR highlighted some molecules that were associated with tumor stages at the mRNA level, such as enolase 1, chromosome 1 ORF 10, elastase inhibitor, α B crystalline, stress-induced phosphoprotein 1, and squamous cell carcinoma antigen 1. Altogether, these data provided further information on ESCC progression and potential drug targets for ESCC clinical therapy.

[Surveillance of cytomegalovirus for antiviral efficacy and risk factors in allogeneic hematopoietic stem cell transplantation].

The study was aimed to investigate the pp65 antigen of human cytomegalovirus (CMV) and its clinical significance in patients revived allogeneic hematopoietic stem cell transplantation (HSCT). 104 patients received allogeneic HSCT were studied. Anticoagulant blood samples were obtained from the recipients before and after transplantation and in the convalescence. CMV pp65 antigen in leukocytes was detected by indirect immunofluorescence assay using CMV Brite Kit weekly. The results showed that among the 104 patients, 29 cases were CMV pp65 positive (27.88%). Out of 29 cases 16 were CMV antigenemia and 13 cases were CMV disease. There were 25 cases who positively responded to antiviral therapy (effective ratio 86.21%) and 4 cases died (case-fatality ratio 13.79%). The detection revealed a significant difference in the incidence of CMV infection between the patients received unrelated or haploidentical family donor HSCT (39.29%) and HLA-identical sibling donor HSCT (14.58%) (P < 0.05). The incidence rate of CMV infection in patients with 0-I grade aGVHD and patients with II-IV grade aGVHD were 19.44% and 46.88% respectively, which had significant difference (P < 0.05). There was significant difference in the occurrence of aGVHD between the patients with and without positive CMV pp65 (P < 0.05). It is concluded that infection of CMV can be detected by the CMV pp65 monoclonal fluorescence immunohistochemistry, The detection of CMV pp65 antigen in peripheral blood leukocytes as a indicator for CMV disease surveillance after HSCT, which may be used to early diagnose the CMV infection, to guide the antiviral treatment and evaluate its efficacy.