Objective To investigate the effect of miR-29c on radiosensitivity of hepatoma HepG2 cells by targeting AKT2 gene.Methods The expression of miR-29c in human normal hepatocytes THLE-3 and hepatoma cell HepG2 was detected by RT-PCR.The relationship between miR-29c and AKT2 were predicted by predicted by informative analysis and verified by dual luciferase reporter gene test and Western blot.miR-29c mimic/AKT2 gene recombinant plasmid and miR-29c inhibitor/ lentivirus vector AKT2 shRNA were transfected into HepG2 cells by Liposome 2000.The cells were irradiated with different doses (0,2,4,6 and 8 Gy) of X-rays,and the effects of miR-29/AKT2 on the survival and cell viability of HepG2 cells were detected by cloning and MTT assays.Results Compared with THLE-3 cells,the expression of miR-29c in HepG2 cells was significantly lower (t=17.816,P＜0.05).After 2,4,6 and 8 Gy X-ray irradiation,the survival of HepG2 cells was significantly lower than that of THLE-3 cells (t =4.541,6.823,7.218,9.363,P＜0.05),and the expression of miR-29c in HepG2 cells was significantly decreased (t =5.599,9.262,10.470,10.873,P＜0.05).The survival and viability of HepG2 cells were decreased by miR-29c overexpression (tsurvival rate =4.307,7.668,7.668,6.894,P＜0.05;tcell viability =3.443,8.116,13.434,P ＜ 0.05) but they were increased by miR-29c inhibition (tsurvival rate =4.003,6.713,7.141,P＜0.05;tcell viability =4.282,5.113,P＜0.05).Double luciferase reporter gene experiments showed that AKT2 was the target gene of miR-29c since the expression of AKT2 was negatively regulated by miR-29c.After the silence of AKT2 or overexpression of AKT2,the survival and viability of HepG2 cells were consistent with the overexpression of miR-29c or the inhibition of miR-29c,respectively.Conclusions MiR-29c increases the radiosensitivity of hepatoma cell HepG2 by targeting AKT2.

Objective To explore the expression of microRNA (miRNA,miR)-449a in liver cancer,the effect on migration and invasion,and the possible regulatory mechanism.Methods Eighteen cases of liver cancer tissues and adjacent tissues were collected.The expression of miR-449a in liver cancer tissues and adjacent tissues,4 kinds of human liver cancer cell lines,and human normal hepatocytes was detected by reverse transcriptase-polymerase chain reaction (RT-PCR).The human liver cancer cell lines were transfected with miR-449a mimic and the negative control (NC) plasmid.The expression of miR-449a after transfection was detected by RT-PCR.The cell number of invasion and migration after transfection in the two groups was measured by Transwell assays.The expression of CXC chemokine ligand 5 (CXCL5) protein in the cells was detected by Western blotting.Results As compared with the adjacent tissues,the expression of miR-449a in liver cancer tissues was significantly reduced (0.957 ±0.165 vs.0.462 ±0.221,t =4.137,P =0.021).As compared with the human normal hepatocytes,the expression of miR-449a in four kinds of human liver cancer cell lines was significantly reduced too (P =0.009,0.008,0.005,0.020),the difference being statistically significant (BEL-7402:6.873 ± 1.293,HepG2:7.246 ± 1.320,NC:1.000,P =0.012,0.017).Two kinds of human liver cancer cell lines with the middle expression of miR-449a were chosen,and the expression of miR-449a after transfection was statistically increased,and the cell number of invasion (BEL-7402 vs.NC:21.57±13.32 vs.79.84± 11.68;HepG2 vs.NC:45.32 ±12.65 vs.92.21 ±15.43,t=24.331,21.109,P=O.011,0.019) and migration (BEL-7402 vs.NC:10.25±3.21 vs.26.63±6.48;HepG2 vs.NC:23.44 ± 10.08 vs.65.65 ± 12.26,t =11.039,12.224,P =0.010,0.008) after transfection was significantly reduced.The expression of CXCL5 proteinafter transfection was statistically significantly reduced (BEL-7402 vs.NC:0.367 ±0.083 vs.0.965 ±0.110;HepG2 vs.NC:0.163 ±0.043 vs.0.949±0.141,t =20.176,25.158) (P=0.021,0.016).Conclusion The miR-449a in liver cancer is expressed weakly,and can serve as a tumor suppressor to influence the migration and invasion of liver cancer cells through targeting CXCL5.

Objective To investigate the protective effect of paeoniflorin on acute pancreatitis with lung injury and its effect on nuclear factor-κB (NF-κB) signaling pathway.Methods Rats were randomly divided into 3 groups (n=26 each): the sham operation group, severe acute pancreatitis (SAP) group, and the paeoniflorin group.After the model of ASP was established, the serum amylase (AMY), interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) expression levels were detected by enzyme linked immunosorbent assay (ELISA).The NF-κB mRNA expression level was detected by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR), and the wet/dry ratio and pathological changes of lung tissue were measured and compared.Results The expression level of AMY [(4 267±1 459), (2 681±822) vs.(1 482±5 240) U/L] in peripheral blood, MPO [(5.47±1.28), (4.39±0.57) vs.(2.17±0.37) U/g] in the lung, and the wet/dry ratio (5.13±0.62, 4.54±0.34 vs.4.41±0.21) in the lung tissue in SAP group and the paeoniflorin group were significantly higher than in the sham operation group at 6 h after the operation, and the changes were even more significant in 12 h sub-groups.Pathological score of pancrea tissue (7.44±1.57 vs.11.30±2.27, P=0.021) and lung tissue (0.83±0.82 vs.1.47±1.31, P=0.029), aortic blood TNF-α [(83.44±15.28) vs.(120.05±25.88) ng/L, P=0.042], IL-1β [(46.33±10.03) vs.(84.75±17.83) ng/L, P=0.036], IL-6 [(72.77±18.51) vs.(92.81±19.92) ng/L, P=0.046] protein levels and NF-κB mRNA in paeoniflorin group were significantly lower than in SAP group 12 h after the operation (P=0.002).Conclusion Peoniflorin may have the positive protection on lung injury in SAP through inhibition of the NF-κB signaling pathway and thus reducing the release of inflammatory factors such as IL-1β, IL-6 and TNF-α.

Objective To improve the anti-tumor effect in vitro and in vivo,a bispecific antibody (CT-BiAb)was constructed.Methods CT-BiAb was obtained by overlapping polymerase chain reaction (PCR) between variable region of Trastuzumab,G4S linker and variable region of Cetuximab.Pichia pastoris was used to express.The binding activity of CT-BiAb to PANC-1 was detected by flow cytometry.Methyl thiazol tetrazolium (MTT) assay was used to detect the inhibition of cell growth.And flow cytometry was used to detect the apoptosis of PANC-1 after treatment with CT-BiAb.Finally,mouse tumor model was generated by endermic injecting PANC-1 cells,and we used this model to detect the antitumor activity of CT-BiAb.Results CT-BiAb was purified and the Western blot result showed that CT-BiAb has a correct molecular weight.CT-BiAb maintained binding activity to PANC-1 and the binding ratio was 50.9%.From the MTT assay,CT-BiAb restrained cell growth.From the apoptosis analysis,we found the cell apoptosis ratio of CT-BiAb (33.50±7.14)%was better than patented Trastuzumab (15.86±4.32)% (P=0.022) and Cetuximab (18.64±5.71)% (P=0.048).In vivo assay of the PANC-1 cell xenografts in nude mice showed that the tumor inhibitory rate of high dose group (73.76±10.21)% was better than patented Trastuzumab (32.55±14.42)% (P=0.001) and Cetuximab (52.63±8.47)% (P=0.006).Conclusion We established and express CT-BiAb,which maintained the binding activity of parent antibody.CT-BiAb provide rationale and drug development strategy for dual inhibition of human epidermal growth factor receptor-2 (Her-2) and epidermal growth factor receptor (EGFR) signaling in pancreatic carcinoma.

Objective To observe the effect of immunotherapy on hepatocellular carcinoma (HCC) by melanoma-associated antigen-A3 (MAGE-A3) activated dendritic cells (DC)-induced antigen-specific CD8+ cytotoxic T lymphocytes (CTL) and offer help to HCC treatment.Methods After MAGE-A3 activated DC biomarkers, interleukin (IL)-12, and IL-10 in DC were tested.MAGE-A3-DC was used to induce antigen-specific CD8+ CTL.The apoptosis of L02 cells and HepG2 cells was detected after treatment with CD8+ CTL.Interferon (IFN)-γ of CD8+ CTL was detected by enzyme-linked immunospot assay (ELISPOT).Subcutaneous tumor model of mouse HCC was established.The tumor volume of different groups was measured.Tumor tissue was further observed by pathological sections.Results MAGE-A3 protein stimulation significantly increased the levels of Human leukocyte antigen-DR (HLA-DR), DC83, DC86 and DC80 markers on the surface of DC (97.45±2.58 vs.48.33±1.68, 92.81±2.36 vs.52.92±1.90, 94.00±3.01 vs.53.97±2.05, 92.16±1.87 vs.34.13±1.32;t=35.680,P=0.013;t=29.440,P=0.021;t=24.580,P=0.012;t=56.690,P=0.009).The IL-12 level of MAGE-A3-DC was significantly higher than that of DC [(338.44±18.15) pg/ml vs.(243.23±16.56) pg/ml;t=8.670,P=0.005], but IL-10 level of MAGE-A3-DC was significantly lower than that of DC [(207.21±10.89) pg/ml vs.(327.58±14.36) pg/ml;t=14.930, P=0.009].The apoptosis of L02 cells treated with MAGE-A3-DC-CD8+ CTL was similar to that after treatment with DC-CD8+ CTL [(9.10±1.40)% vs.(9.71±1.58)%;t=0.650, P=0.120].The apoptostic rate of HepG2 cells treated with MAGE-A3-DC-CD8+ CTL was significantly higher than that after treatment with DC-CD8+ CTL [(58.84±5.27)% vs.(9.63±1.61)%;t=19.970, P=0.008].After therapy, tumor sizes V/V0 in MAGE-A3-DC-CD8+CTL group decreased significantly as compared with those of phosphate buffer (PBS) group and DC-CD8+ CTL group (5.43±1.22 vs.21.81±2.01 vs.22.85±2.40;t=22.030, P=0.010;t=20.460, P=0.012).Hematoxylin and eosin (HE) staining showed that nuclear condensation, increased cell gap, vacuoles, and edema were observed in tumor tissues after treatment with MAGE-A3-DC-CD8+ CTL, but pathological changes of tumor tissues were not observed in PBS group and DC-CD8+ CTL group.Conclusion MAGE-A3 protein can stimulate the maturation of DC that induce the production of MAGE-A3 specific CD8+CTL.MAGE-A3-DC-CD8+CTL have specific tumor killing ability.

Objective To investigate the clinical significance and interrelation of long-chain non-coding RNA (lcnRNA) HOXA transcript at the distal tip (HOTTIP) and metabotropic glutamate receptor 1 (mGluR1) in pancreatic cancer patients.Methods Pancreatic cancer samples,which were treated in our hospital were collected.The expression levels of HOTTIP and mGlUR1 mRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in fresh pancreatic cancer tissue,and those of HOTTIP and mGluR1 protein by Western blotting and immunohistochemistry.Moreover,the relationship between the expression of HOTTIP,mGlUR1 and clinicopathological features was analyzed.Results The levels of HOTHP and mGlUR1 mRNA and protein were significantly up-regulated in pancreatic cancer tissues as compared with normal pancreatic tissue (P =0.000).The expression of HOTTIP protein was positive in 32 samples (68.09％) and mGlUR1 protein was positive in 30 samples (63.83％).The positive rate of HOTTIP and mGlUR1 in adjacent tissues was 31.91％ and 36.17％,respectively,and the difference was also statistically significant (P =0.000).The expression of HOTTIP and mGluR1 was correlated with lymph node metastasis,differentiation degree and TNM stage (P =0.000,0.004,0.000).The survival time of HOTTIP and mGlUR1 positive patients was significantly shorter than that of negative patients (P =0.029,0.047).Cox regression multivariate analysis showed that lymph node metastasis,differentiation degree,TNM stage,HOTTIP and mGlUR1 expression were independent prognostic factors for pancreatic cancer (P =0.022,0.048,0.026,0.000,0.002).In addition,HOTTIP and mGlUR1 expression showed a significant positive correlation in cervical cancer patients (r =0.689,P =0.002).Conclusion The positive expression of HOTTIP and mGlUR1 is associated with the occurrence,progression and prognosis of pancreatic cancer.

Runt-related transcription factor 2 (RUNX2) was first identified as a transcription factor to play an important role in different biological processes of osteoblast and chondrocyte, including differentiation and migration. Recently, RUNX2 has been implicated in promigratory/proinvasive behavior in different human malignancies. In the present study, we demonstrated that the RUNX2 mRNA and protein expression were both increased significantly in HCC tissues and cell lines. High RUNX2 expression was correlated obviously with poor clinicopathological characteristics including multiple tumor nodes, high histological grading, venous infiltration and advanced tumor-node-metastasis (TNM) stage. In addition, we demonstrated that RUNX2 was a prognostic indicator for predicting 5-year overall survival and disease-free survival of HCC patients. Our studies showed that RUXN2 overexpression promoted, while RUNX2 knockdown inhibited HCC cell migration and invasion in vitro. Notably, RUNX2 positively regulated matrix metalloproteinase 9 (MMP9) accumulation in HCC cells. Furthermore, we confirmed that RUNX2 was positively correlated with MMP9 expression in HCC tissues by Pearson correlation analysis. Mechanistically, we demonstrated that MMP9 overexpression increased HCC cell migration and invasion, while MMP9 knockdown reduced HCC cell migration and invasion in vitro. Alteration of MMP9 expression partially abrogated the effects of RUNX2 on HCC cell migration and invasion, which suggests that RUNX2 developed its pro-metastatic biological function by upregulating the expression of MMP9 in HCC cells. In conclusion, our results reveal that RUNX2 promotes HCC cell migration and invasion by MMP9-mediated pathway, and potentially serves as a new prognostic biomarker and in therapeutic strategies for HCC.

Background: MicroRNAs (miRNAs) play critical roles in hepatocellular carcinoma (HCC) development and progression. Aberrant miR-21 expression has been reported in several cancers. However, the clinical significance of miR-21 in human HCC is still unclear. Methods: A total of 112 patients with primary HCC who underwent a curative liver resection were included in this retrospective study. The differentially expressed amount of the miR-21 was validated by quantitative real-time PCR (qRT-PCR). Survival rate was analyzed by log-rank test, and survival curves were plotted according to Kaplan-Meier. Multivariate analysis of the prognostic factors was performed with Cox regression model. Results: As revealed by qRT-PCR analysis, miR-21 expression was significantly upregulated in HCC tissues when compared with adjacent non-tumor tissues (P<0.05). High miR-21 expression level was observed to be closely correlated with tumor differentiation, TNM stage and vein invasion (P<0.05). Patients who had high miR-21 expression had a shorter overall survival than patients who had low miR-21 expression (P<0.05). Moreover, multi-variate analysis of the prognosis factors with a Cox proportional hazards model showed that high miR-21 expression was a significant independent predictor of poor survival in HCC (P<0.05). Conclusion: Our results suggested that increased expression of miR-21 was significantly correlated with tumor progression and could be a novel potential biomarker for HCC prognosis.

Introduction: Cancer stem cells (CSCs) are believed to be ` seed cell' in cancer recurrence and metastasis. MicroRNAs (miRNAs) have emerged as potential therapeutic candidates due to their ability to regulate multiple targets involved in tumor progression and chemoresistance. The goal of this study was to investigate the role of miRNA-200c (miR-200c) in regulating colony formation, invasion and chemoresistance of human pancreatic cancer stem cells (PCSCs). Methods: PCSCs with CD24(+) CD44(+) ESA(+) as the marker was sorted from PANC-1 cell line by fluorescence activated cell sorter (FACS). Quantitative real-time PCR (qRT-PCR) assay was used to detect the expression of miR-200c in PCSCs and PANC-1 cells. Transfection of miR-200c mimic into PCSCs was performed to establish miR-200c over-expressed cells. The effects of overexpressing miR-200c on PCSCs were examined by cell colony forming, invasion and survival assays in vitro. Results: Our data showed that CD24(+) CD44(+) ESA(+) PCSCs (0.5%) were isolated from PANC-1 cells. Expression of miR-200c was significantly reduced in PCSCs compared with PANC-1 cells. In addition, the capability of colony formation, invasion and chemoresistance were markedly increased in PCSCs than that in PANC-1 cells. Adverse results were obtained in miR-200c overexpressing PCSCs transfected with miR200c mimic. Conclusion: Our study demonstrated that miR-200c overexpression could decrease colony formation, invasion and chemoresistance of PCSCs. It may become a new therapeutic target for gene therapy in patients suffered from pancreatic cancer.

Objective To explore the therapeutic effect of wortmannin on acute pancreatitis (AP) with renal dysfunction and on phosphatidylinositol 3 kinase (PI3K)/protein kinase B (PKB) signaling pathway.Methods Eighty healthy male SD rats were randomly divided into normal control group,sham operation group (SO group),AP group and wortmannin group.SO group,AP group and wortmannin group were further divided into 3,6 and 12 h sub-groups (n =8 each).The AP model was established by bile duct retrograde injection.Wortmannin was intraperitoneally injected 4 h before operation.Serum urea nitrogen,creatinine,amylase,pathological findings of the kidney and pancreas,and PKB,p-PKB and tumor necrosis factor-α (TNF-α) proteins in the kidney and pancreas were observed.Results Histopathological damage of the kidney and pancreas was severer,blood urea nitrogen [(9.81 ± 1.28) and (77.49±1.17) vs.(5.33 ±0.32) mmol/L],creatinine [(62.19 ±5.84) and (55.12 ±5.27) vs.(45.13 ±3.01) μmol/L] and amylase levels [(2 931 ±619) and (2 061 ±897) vs.(677 ± 120) U/L] were significantly higher in AP group and wortmannin group than in SO group 3 h after operation (P ＜ 0.05),more significantly in 6 h and 12 h sub-groups.Histopathological damage of the kidney and pancreas was severed,blood urea nitrogen [(16.51 ± 2.00) vs.(21.16 ± 2.57) mmol/L],creatinine [(94.63±11.30) vs.(116.24± 14.82) μmol/L],amylase levels [(3264±932) vs.(7725± 1 517) U/L],p-PKB (1.01 ± 0.24 vs.1.23±0.30) andTNF-αproteins (1.11±0.29vs.1.33± 0.37) levels were significantly lower in wortmannin group than in AP group at 12 h after operation (P ＜ 0.05).Conclusion Activation of PI3K/PKB signaling pathway plays a very important role in the occurrence and development of AP with renal function dysfunction.PI3K inhibitors (wortmannin) can significantly alleviate the renal function damage induced by AP.

Objective To explore the effect of microRNA (miRNA,miR)-200c in reversing chemo-resistance of human pancreatic cancer stem cells.Methods CD24 + CD44 + ESA + cells were sorted from PANC-1 cell line by FACS.miR-200c precursor fragment and negative control fragment were transfected into pancreatic cancer stem cells.The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect the miR-200c expression.MTT assay was used to measure drug sensitivity to gemcitabine.Results CD24+ CD44 + ESA + cells (0.8％) were isolated in PANC-1 cells.The relative miR-200c expression level in pancreatic cancer stem cell line (0.15 ± 0.01) was significantly lower than that in PANC-1 cell line (1.00 ± 0.09,P ＜ 0.05).The half inhibition concentration (IC50) of gemcitabine was significantly higher in pancreatic cancer stem cells group [(19.15 ± 1.53) μmol/L] than that in PANC-1 cells group [(0.86 ±0.18) μmol/L] (P＜0.05).The relative miR-200c expression level in pancreatic cancer stem cell line in miR-200c precursor transfected group [(3.70 ± 0.42)] was significantly higher than that in negative control group (0.22 ±0.21,P ＜0.05).The IC50 of gemcitabine was significantly lower in miR-200c precursor transfected group [(0.92 ± 0.13) μ mnol/L] than that in negative control group [(17.94 ± 1.36) μmol/L,P ＜ 0.05].Conclusion miR-200c could reverse chemo-resistance of human pancreatic cancer stem cells to gemcitabine.

Objective To investigate the sensitivity of human pancreatic cancer stem cells to chemotherapy and the expression of microRNA (miRNA,miR)-200c in cancer stem cells.Methods CD24 + CD44 + ESA + cells were sorted from PANC-1 cell line by fluorescence-activated cell sorter (FACS).The stem like properties of this subpopulation were assessed by non-obese diabetic (NOD)/ severe combined immunodeficiency (SCID) xenograft transplantation experiment.Sensitivity to gemcitabine and apoptosis ratio of pancreatic cancer stem cells and PANC-1 cells were detected by methyl thiazol tetrazolium (MTT) assay and FACS respectively.The real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect miR-200c expression in pancreatic cancer stem cells and PANC-1 cells.Results CD24 + CD44 + ESA + cells (0.8％) were isolated in PANC-1 cells.NOD/SCID xenografl transplantation experiment confirmed that the sub-group had the characteristics of cancer stem cells.The half inhibition concentration (IC50) of gemcitabine was significantly higher in pancreatic cancer stem cells group [(19.15 ± 1.53) μmol/L] than in PANC-1 cells group [(0.86 ± 0.18) μmol/L] (P ＜ 0.05).Mter the interference of gemcitabine (1,10 μmol/L),the apoptosis ratio was significantly lower in pancreatic cancer stem cells group [(0.69 ±0.09)％ and (0.90 ±0.13)％] than that in PANC-1 cells group [(60.54 ± 3.73) ％ and (91.76 ± 5.07) ％] (P ＜ 0.05).The relative miR-200c expression level in cancer stem cell line [(0.15 ±0.01)] was significantly lower than that in PANC-1 cell line [(1.00 ± 0.09)] (P ＜ 0.05).Conclusion The expression of miR-200c was significantly reduced in pancreatic cancer stem cells which were significantly chemo-resistant,and miR-200c may play an important role in the chemo-resistance of pancreatic cancer stem cells.

Objective:To investigate the expression and action of microRNA-200c (miRNA-200c) in pancreatic cancer stem cells. Methods:Pancreatic cancer stem cells were sorted from human pancreatic cancer PANC-1 cells by FACS using CD24+CD44+ESA+as marker, and their stem cell properties were assessed by xenogratf tumor assay in NOD/SCID mice. hTe miRNA-200c expression and invasion ability in PANC-1 cells, pancreatic cancer stem cells and pancreatic cancer stem cells transfected with miRNA-200c precursor sequence or negative control sequence were determined by RFQ-PCR method and Transwell invasion assay, respectively. Results:The CD24+CD44+ESA+cells (accounting for 0.8%) sorted from PANC-1 cells presented tumor stem cell properties, and the volume of the xenograft tumor after their subcutaneous transplantation in mice was significantly larger than that after PANC-1 cells transplanted at the same time [(1 725.14±261.29) mm3 vs. (479.65±99.67) mm3, P<0.05]. In pancreatic cancer stem cells compared with PANC-1 cells, the miRNA-200c expression level was significantly decreased (0.15±0.01 vs. 1.00±0.09, P<0.05), and transmembrane cell number was signiifcantly increased (321±7.62 vs. 70±16.47, P<0.05), but the miRNA-200c expression level was signiifcantly increased and transmembrane cell number was signiifcantly decreased in pancreatic cancer stem cells atfer transfection with miRNA-200c precursor sequence (both P<0.05). Conclusion:MiRNA-200c expression is reduced in pancreatic cancer stem cells, and miRNA-200c has inhibitory effect on growth and invasiveness of pancreatic cancer stem cells.

Invasion and metastasis is the main causes leading to the death of hepatocellular carcinoma (HCC) patients. However, the underlying mechanism is still to be explored. Transforming growth factor β1 (TGF-β1) is a stronger inducer of HCC cell invasion. However, the downstream effector of TGF-β1 that promotes HCC invasion is still unknown. In this study, we found that PI3K/Akt activation takes place following the stimulation of TGF-β1. The inhibition of PI3K/Akt activation abolished epithelial-mesenchymal transition (EMT) and invasion of HCC cells induced by TGF-β1. Myocyte enhancer factors 2 (MEF2) family proteins were found to be overexpressed in HCC cells under the treatment of TGF-β1 in a PI3K/Akt-dependent way. Silencing the expression of MEF2s was able to prevent the effect of TGF-β1 on HCC EMT and invasion. Unexpectedly, MEF2 proteins were able to promote the expression of TGF-β1 in HCC cells, suggesting the existence of regulatory circuitry consisting of TGF-β1, PI3K/Akt, and MEF2. A natural compound, oleanolic acid, was demonstrated to suppress the invasion and EMT of HCC cells by downregulating MEF2, showing that targeting this pathway is an effective therapeutic strategy for HCC invasion. We believe that our findings can contribute to better understanding of the involved mechanism of HCC invasion and the development of preventive and therapeutic strategy.

P2X7 is important in inflammation and tissue injury. The aim of the present study was to investigate the effect of P2X7 inhibition, using a specific inhibitor (A438079) to prevent the development of liver injury and fibrosis in a mouse model of liver fibrosis. The mouse liver fibrosis model was induced by carbon tetrachloride (CCl4). Mice received subcutaneous administration of vehicle (saline/olive oil), CCl4 or subcutaneous CCl4 and A438079. The pro‑inflammatory and pro‑fibrotic factors were determined by western blot analysis. The biochemistry, histopathology, collagen deposition and nuclear factor‑κB (NF‑κB) activity were also analyzed. Chronic CCl4 treatment resulted in liver injury and collagen accumulation. The expression levels of P2X7, pro‑inflammatory and pro‑fibrotic mediators, and the activity of NF‑κB were markedly increased. Treatment with A438079 significantly inhibited CCl4‑induced P2X7 expression, and attenuated CCl4‑induced liver injury and the inflammatory response. P2X7 blockade also significantly reduced the formation of collagen in the liver and the expression of α-smooth muscle actin and transforming growth factor‑β1. This study demonstrated that P2X7 inhibition attenuated liver injury and fibrosis in a mouse model. Thus, P2X7 is a potential novel therapeutic target for liver injury and fibrosis.

Introduction: Dysregulation of long non-coding RNAs (lncRNAs) plays critical roles in tumor progression. lncRNA LOC285194 was previously shown to be correlated with aggressive clinicopathological features and poor prognosis in several cancers. The aim of this study was to investigate relationship between LOC285194 expression and clinical outcomes in human pancreatic ductal adenocarcinoma (PDAC). Methods: Quantitative real-time PCR (qRT-PCR) assay was performed to detect the expression of lncRNA LOC285194 in human PDAC cells and tissue samples. The association of LOC285194 expression with clinicopathologic features was analyzed. Kaplan-Meier analyses were used to assess survival of patients. Univariate and multivariate analyses were performed using the Cox proportional hazards model to analyze the prognostic significance of LOC285194 expression. Results: Our data showed that the relative level of LOC285194 in PDAC cells was significantly lower than that in normal human pancreatic duct epithelial cell line. Also, the expression of LOC285194 in PDAC tissues was significantly lower than that in adjacent non-tumor tissues. By statistical analyses, low LOC285194 expression was observed to be closely correlated with clinical stage, lymphnode metastasis and liver metastasis. Kaplan-Meier survival analysis revealed that patients with low LOC285194 expression had a poor overall survival compared with the high LOC285194 group (P < 0.05). Univariate and multivariate analyses showed that low LOC285194 expression was an independent poor prognostic factor for PDAC patients. Conclusions: Our data provided the first evidence that reduced LOC285194 in PDAC tissues was correlated with tumor progression, and lncRNA LOC285194 might be a potential molecular biomarker for predicting the prognosis of patients.