A radiomics-based model to differentiate glioblastoma from solitary brain metastases.

AIM: To differentiate glioblastoma (GBM) from solitary brain metastases (MET) using radiomic analysis. MATERIALS AND METHODS: Two hundred and fifty-three patients with solitary brain tumours (157 GBM and 98 solitary brain MET) were split into a training cohort (n=178) and a validation cohort (n=77) by stratified sampling using computer-generated random numbers at a ratio of 7:3. After feature extraction, minimum redundancy maximum relevance (mRMR) and the least absolute shrinkage and selection operator (LASSO) were used to build the radiomics signature on the training cohort and validation cohort. Performance was assessed by radiomics score (Rad-score), receiver operating characteristic (ROC) curve, calibration, and clinical usefulness. RESULTS: Eleven radiomic features were selected as significant features in the training cohort. The Rad-score was significantly associated with the differentiation between GBM and solitary brain MET (p<0.001) both in the training and validation cohorts. The radiomics signature yielded area under the curve (AUC) values of 0.82 and 0.81 in the training and validation cohorts to distinguish between GBM and solitary brain MET. CONCLUSIONS: The radiomics model might be a useful supporting tool for the preoperative differentiation of GBM from solitary brain MET, which could aid pretreatment decision-making.

GPAA1 promotes progression of childhood acute lymphoblastic leukemia through regulating c-myc.

OBJECTIVE: Previous studies have shown that glycosylphosphatidylinositol Anchor Attachment Protein 1 (GPAA1) is a cancer-promoting gene; however, the role of GPAA1 in childhood acute lymphoblastic leukemia (ALL) has not been reported. This study aims to illustrate the role of GPAA1 in promoting the metastasis of ALL by targeting c-myc and the potential mechanism. PATIENTS AND METHODS: Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to examine serum levels of GPAA1 and c-myc in 42 childhood ALL patients and healthy volunteers. The interaction between GPAA1 expression and prognosis of childhood ALL was analyzed. Meanwhile, expressions of GPAA1 and c-myc in ALL cell lines were determined by qRT-PCR. Furthermore, after GPAA1 knockdown model was constructed by lentivirus transfection in MOLT-4 and SUP-B15 cells, cell counting kit-8 (CCK-8), transwell invasion, and cell wound healing assays were conducted to analyze the effect of GPAA1 on the biological functions of ALL cells. Potential mechanism was further explored through Luciferase reporter gene assay and cell recovery experiments. RESULTS: QRT-PCR results indicated that serum level of GPAA1 in childhood ALL patients was remarkably higher than that of healthy volunteers, and the difference was statistically significant. Childhood ALL patients with high expression of GPAA1 had lower overall survival rate compared with those expressing low expression of GPAA1. Proliferation and metastasis abilities of pediatric ALL cells with GPAA1 knockdown remarkably decreased. Subsequently, c-myc expression was also found remarkably upregulated in ALL cell lines and serum samples of childhood ALL patients and it was positively correlated with GPAA1 level. In addition, Luciferase reporter gene assay demonstrated that overexpression of c-myc remarkably attenuated the Luciferase activity of the wild-type GPAA1 vector without attenuating that of the mutant vector or empty vector, further demonstrating that GPAA1 can be targeted by c-myc. At the same time, cell recovery experiment found that the interaction between GPAA1 and c-myc together regulated the malignant progression of ALL. CONCLUSIONS: GPAA1 was up-regulated in serum of childhood ALL patients, which was remarkably associated with the prognosis. In addition, GPAA1 may contribute to the malignant progression of childhood ALL via activating c-myc.

Effect of NLK on the proliferation and invasion of laryngeal carcinoma cells by regulating CDCP1.

OBJECTIVE: The morbidity and mortality of laryngeal cancer are increasing rapidly, seriously threatening human health. There are several causes of laryngeal cancer, but the exact molecular mechanism is unclear. Finding the molecular targets of laryngeal cancer has become an emerging hot spot. Nemo-like kinase (NLK) is abnormally expressed in tumors, but its role in laryngeal cancer has not been reported. PATIENTS AND METHODS: Real Time PCR and Western blot were used to detect NLK mRNA and protein expression in cancer tissues and adjacent tissues of laryngeal cancer patients. The laryngeal carcinoma cell line Hep-2 cells were cultured in vitro and randomly divided into three groups: control group, NC group, and NLK siRNA group followed by the analysis of cell proliferation by MTT assay, Caspase3 activity, and cell invasion by the transwell chamber. MMP-9 and CDCP1 expression was measured by Western blot. RESULTS: NLK mRNA and protein expression was significantly increased in laryngeal carcinoma tissues compared with those in adjacent tissues (p<0.05). NLK siRNA transfection into Hep-2 cells significantly down-regulated NLK expression, inhibited Hep-2 cell proliferation and invasion, increased Caspase-3 activity with statistical differences compared to control group (p<0.05). Down-regulation of NLK expression in Hep-2 cells inhibited MMP-9 expression and decreased CDCP1 expression. CONCLUSIONS: NLK is expressed in tumor tissues of patients with laryngeal cancer. The down-regulation of NLK expression may play a role in the proliferation, apoptosis, and invasion of laryngeal carcinoma cells and it is possible by regulating MMP-9 and CDCP1 expression.

CT and MRI studies of hepatic angiosarcoma.

AIM: To analyse the computed tomography (CT) and magnetic resonance imaging (MRI) manifestations of hepatic angiosarcoma. MATERIALS AND METHODS: Nineteen patients with hepatic angiosarcoma underwent preoperative abdominal unenhanced and contrast-enhanced CT (11 cases) or (eight cases) MRI. RESULTS: The results of a coagulation examination showed varying degrees of abnormalities in 12 (63.16%) cases (most were prolonged prothrombin time and an increased proportion of prothrombin time), which were the most common abnormalities on the laboratory tests. Unenhanced CT of the lesions showed homogeneous or heterogeneous hypointense with hyperintense haemorrhagic lesions, contrast-enhanced CT was performed for six cases (6/11, 54.5%) with centripetal heterogeneous filling. All lesions showed heterogeneous intensity at MRI, including heterogeneous hypointense T1WI and homogeneous or heterogeneous hyperintense T2WI. Haemorrhage lesions showed higher hyperintensity with spot or patchy signals. Centripetal enhancement was found in six cases using contrast-enhanced imaging. Flaky patches of contrast enhancement were seen in the lesions. CONCLUSION: The CT and MRI features of most of the hepatic angiosarcomas in the present study were relatively characteristic: the border of the mass was indistinct, the density was heterogeneous, and haemorrhage was frequently seen, with secondary calcification in a few cases, whereas enhanced imaging showed typical centripetal heterogeneous enhancement. In addition, highly malignant angiosarcoma could not be enhanced.

Transforming Growth Factor-ß Partially Reversed the Immunosuppressive Effect of Mesenchymal Stem Cells in Mice.

BACKGROUND: Mesenchymal stem cells (MSCs) possess powerful immunosuppression capacity. Transforming growth factor-ß (TGF-ß) is a well-known anti-inflammatory cytokine and plays an important role in various inflammatory processes. We hypothesized that TGF-ß could synergize with MSCs in suppressing immune responses, and therefore established a mouse skin graft model to evaluate the effect of MSCs and MSCs combined with TGF-ß on transplantation immunity in vivo. METHODS: Balb/c and C57BL/6 mice were used to establish the skin graft model. The recipients were divided into 3 groups and received intravenous bone marrow mesenchymal stem cells (BMSCs), BMSCs pretreated with TGF-ß, and 0.9% saline solution, respectively. Skin graft survival time, pathological detection, the ratio of CD4+CD25+Foxp3+Treg cell of spleens, and the level of IFN-γ, IL-2, IL-10, and TGF-ß expression were tested. RESULTS: The survival time of skin grafts were prolonged in both BMSC (12.5 ± 1.35 days) and BMSC-TGF-ß (10.6 ± 1.90 days) recipients compared to the blank control recipients (8.0 ± 1.05 days). The ratio of CD4+CD25+Foxp3+Treg cell of spleens from BMSC and BMSC-TGF-ß recipients was higher than that of the blank control, and the upregulated proliferation in the BMSC group occurred earlier and was prolonged compared to the BMSC-TGF-ß group. The expression of IFN-γ and IL-2 was inhibited in both the BMSC and BMSC-TGF-ß groups compared to the blank, while the expression of IL-10 and TGF-ß was boosted. In contrast to the BMSC group, the BMSC-TGF-ß group exhibited a weaker effect on the expression of cytokines. CONCLUSION: TGF-ß partially reversed the immunosuppressive effect of MSCs in vivo. This immunoregulatory feature may have potential applications for treating transplant rejection.

Variations of antibiotic resistance profiles in chickens during administration of amoxicillin, chlortetracycline and florfenicol.

AIM: To assess the effect of antibiotics administered in feed on the resistance phenotypes and genotypes of Escherichia coli in the chicken intestine. METHOD AND RESULTS: Chickens were administered amoxicillin, chlortetracycline and florfenicol in feed and 203 intestinal E. coli were examined for their susceptibility to 11 antimicrobial agents and for the presence of antibiotic resistance genes (ARG) using PCR. DNA was extracted from chicken stool samples in 15, 20, 30 and 40 day old chickens. We found that while antibiotic resistance rates increased with time, the relative gene abundance of tet(W), tet(A), cmlA, cfr and sul1 decreased. In contrast, the relative abundance of gene blaTEM and mcr-1 increased over the experimental period. Pearson correlation analysis indicated that sul1 was correlated with tet(W) (R = 0·630, P < 0·01) and cmlA was correlated with cfr (R = 0·587, P < 0·01). Interestingly, mcr-1 correlated with tet(W) (R = -0·546, P < 0·05). CONCLUSIONS: Administration of different antibiotic reduced the relative abundance of ARG in chickens but did not halt the expansion of antibiotic resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: Changing the pattern of antibiotic types used to prevent antibiotic resistance in chickens is not a viable method to prevent the spread of ARG.

Risk factors contributing to early and late recurrence of haemoptysis after bronchial artery embolisation.

OBJECTIVE: To identify the risk factors for early and late relapse of haemoptysis after bronchial artery embolisation (BAE). DESIGN: We performed a retrospective study of 255 patients with haemoptysis who underwent BAE from January 2009 to June 2016 at the First Affiliated Hospital of Nanjing Medical University, Nanjing, China. Using a Cox regression model, risk factors contributing to early (within the first month) and late (>1 month) recurrence were analysed. We censored recurrence-free patients at 1 month and patients without rebleeding after 1 month. RESULTS: The cumulative recurrence rate at 1, 6, 12, 24 and 48 months was respectively 14.9%, 16.9%, 20.6%, 31.0% and 39.9%, with a median follow-up of 574 days. Early relapse occurred in 38 of 255 patients, while late recurrence was found in 45 of the remaining 217 patients. The risk factors related to early recurrence were lung destruction on computed tomography of the chest and involvement of non-bronchial systemic arteries. The risk factors associated with late recurrence were tuberculosis sequelae, the presence of shunts and the use of gelfoam as an embolisation material. CONCLUSIONS: The variables related to incomplete embolisation were the risk factors for early rebleeding. The variables associated with same-vessel recanalisation and formation of a new collateral circulation influenced late recurrence.

The effect of curcumin on cell adhesion of human esophageal cancer cell.

OBJECTIVE: Esophageal cancer is the 8th most common cancers worldwide and the 6th most common cause of death among cancers. Curcumin has been reported to have the function of anti-inflammatory, antioxidant, anti-rheumatoid, and anti-atherosclerosis role. It can also reduce lipid, eliminate free radicals and inhibit the growth of the tumor. Many reports had suggested that curcumin has shown great potential in the treatment of tumors by inducing apoptosis. Little is known about the effects of curcumin on cell adhesion of tumor cancer. Therefore, in this study, we attempted to look for a new approach to target resistant cells and improve efficacy without toxicity. MATERIALS AND METHODS: Human esophageal cancer cell line (Eca-109 cells) was cultured. Cell adhesion was detected under a microplate reader. Reactive oxygen species were measured using Fluostar Omega Spectrofluorimeter. SOD activity and GSH content in cells were detected by commercial determination kit. The expression of p-JAK, p-STAT3 and STAT3 were measured by Western blot and RT-PCR. RESULTS: Cell adhesion assay showed curcumin enhances cell-cell adhesion and cell-matrix adhesion in Eca-109 cells. ROS levels, SOD activity and total GSH content were detected and the results showed curcumin decreases intracellular ROS levels but increases SOD activity and total GSH content. Then, NAC (ROS inhibitor) and ICI (ER inhibitor) were pre-treated. Results showed ICI reversed the decreasing of intracellular ROS levels and the increasing of SOD activity and total GSH content affected by curcumin, but NAC had no such impact. Taken together, ER rather than ROS involves in cell adhesion affected by curcumin. Meanwhile, the downregulating of p-JAK, p-SATA3 and total STAT3 were caused by curcumin but NAC had no such influence. They were reversed by ICI, but NAC had no such influence. CONCLUSIONS: Curcumin could increase cell adhesion through inhibiting JAK/STAT3 mediated by ER in Eca-109.

MicroRNA-124 inhibits cell invasion and epithelial-mesenchymal transition by directly repressing Snail2 in gastric cancer.

OBJECTIVE: MicroRNAs (miRNAs) act as critical regulators of genes expression involved in tumor biological processes. The study aimed to investigate the clinical significance and biological role of miR-124 in gastric cancer (GC). PATIENTS AND METHODS: MiR-124 expression was analyzed from 88 GC tissues and adjacent normal tissues by quantitative Real-time PCR (qRT-PCR). Kaplan-Meier curves and log-rank test was used to evaluate the association between miR-124 and the over survival (OS) time of GC patients. MTT assays and transwell invasion assays were performed to assess cell proliferation and invasion. The relationship between miR-124 and Snail2 expression was analyzed by dual luciferase reporter assay. Western blot analyses were performed to detect the relative protein expression. RESULTS: We found that miR-124 expression was significantly reduced in GC tissue samples when compared to the adjacent normal tissues (p<0.05). Lower miR-124 was found to be associated with tumor size (p=0.001), lymphatic metastasis (p=0.008) and TNM stage (p=0.015). Furthermore, patients who have lower miR-124 predicted poor OS time (p<0.05). Function studies suggested that cell proliferation and invasion ability of GC cells were inhibited by up-regulation of miR-124 expression. Moreover, we demonstrated that Snail2 was a direct target of miR-124. Meanwhile, miR-124 inhibited Epithelial-Mesenchymal Transition (EMT) process by repressing the Snail2 expression in GC cells. CONCLUSIONS: MiR-124 acted as a tumor suppressor in GC and may be a useful target for GC treatment.

Elevated serum retinol-binding protein 4 levels are correlated with blood pressure in prehypertensive Chinese.

Retinol-binding protein 4 (RBP4) has been reported involving in the occurrence and development of hypertension. However, to date, few data are available on the correlation between serum RBP4 level and blood pressure (BP) in prehypertension. Therefore, this association was investigated in prehypertensive Chinese. Overall, 160 subjects with prehypertension (Pre-HT group) and 160 subjects with normal BP (NBP group) were recruited in this study. The subjects were divided into the following four subgroups according to body mass index (BMI): obese Pre-HT subgroup; non-obese Pre-HT subgroup; obese NBP subgroup; and non-obese NBP subgroup (n=80 in each). Anthropometric parameters, systolic BP (SBP), diastolic BP (DBP) and several biochemical parameters were measured. Fasting insulin was evaluated by radioimmunoassay. Serum RBP4 level was measured by enzyme-linked immunosorbent assay. The Pre-HT group had higher levels of serum RBP4 level than did the NBP group (P<0.001). Moreover, higher RBP4 levels were identified in the obese Pre-HT subgroup relative to the non-obese Pre-HT subgroup (P=0.005). However, no difference in RBP4 level was identified between the obese and the non-obese NBP subgroups (P=0.317). RBP4 level was positively correlated with BMI (r=0.226, P=0.001), SBP (r=0.468, P<0.001) and DBP (r=0.358, P<0.001) after adjustment for age, sex, smoking status and alcohol consumption. The results of the multiple regression analyses demonstrated that RBP4 level was independently associated with SBP (ß=0.427, P<0.001) and DBP (ß=0.338, P<0.001). In conclusion, serum RBP4 level was significantly higher and closely associated with BP in prehypertensive Chinese.

Efficacy of a combination of taurine and stenosis removing on cognitive impairment induced by carotid artery stenosis in rats.

OBJECTIVE: We assessed the protective effect on a carotid artery stenosis rat model with using a combination of taurine and stenosis treatment. Rats were randomly divided into five groups: Group 1 (sham), Group 2 (model), Group 3 (stenosis removing), Group 4 (taurine), and Group 5 (combination). MATERIALS AND METHODS: The carotid artery stenosis model was created by ligating the common carotid artery along with various gauges of needle and then removing the needle. The cognitive performance was analyzed using P300 latency and escape latency in the Morris water maze (MWM) task. The levels of oxidative products malondialdehyde (MDA), inflammatory factors interleukin (IL-1ß) and tumor necrosis factor (TNF-α), the activities of anti-oxidative enzymes superoxide dismutase (SOD) and catalase (CAT) were determined using enzyme-linked immunosorbent assay (ELISA) kits, respectively. RESULTS: The results revealed that the cognitive impairment can be reduced by stenosis removing and taurine, combining the administration of stenosis removing and taurine had a significant effect on the recovery of cognitive function. CONCLUSIONS: A further study showed that, the protection of cognitive function is related to inhibition of oxidative stress and suppression of inflammatory response.

Meta-analysis of the association between PADI4 -92C/G polymorphism and rheumatoid arthritis in the Chinese population

Many studies have evaluated the correlation between peptidylarginine deiminase 4 (PADI4) -92C/G polymorphism and rheumatoid arthritis (RA), but the results remain inconclusive. Therefore, we performed a meta-analysis in the Chinese population to provide comprehensive data on the association between PADI4 -92C/G polymorphism and RA. Eligible studies published before May 2016 were identified in PubMed and Chinese databases. The strengths of these associations were assessed by pooled odds ratios (OR) and 95% confidence interval (CI). Eight studies documenting a total of 1351 RA cases and 1585 controls were included in this meta-analysis. In the overall analysis, a significant association between the PADI4 -92C/G polymorphism and RA was found in the Chinese population (G vs C: OR=1.32, 95%CI=1.02-1.71; GG+CG vs CC: OR=1.75, 95%CI=1.20-2.53). The subgroup analyses stratified by geographic area(s) and source of controls revealed significant results in South China, in hospital-based studies and population-based studies. In summary, this meta-analysis suggested that PADI4 -92C/G polymorphism may be associated with the RA incidence in the Chinese population, especially for South China. Further studies conducted on other ethnic groups are required for definite conclusions.

MicroRNA-132 inhibits migration, invasion and epithelial-mesenchymal transition by regulating TGFß1/Smad2 in human non-small cell lung cancer.

OBJECTIVE: Increasing evidence shows that microRNA involves in the development of several types of cancers, however, the role of microRNA-132 (miR-132) in non-small cell lung cancer (NSCLC) metastasis remains largely unknown. In this study, we aimed to investigate the effect of miR-132 on the epithelial-mesenchymal transition (EMT) and the potential mechanisms in NSCLC. PATIENTS AND METHODS: The Quantitative real-time PCR (QRT-PCR) was used to detect the miR-132 levels in 15 NSCLC tissues and cell lines. Transwell and wound healing assays were used to evaluate the function of miR-132 in NSCLC cell metastasis. EMT-related markers were determined by using qRT-PCR. EMT-related TGFß1/Smad2 signaling pathway was explored using Western blot. RESULTS: MiR-132 expression level was lower in NSCLC tissues compared with the matched adjacent normal tissues. It was also downregulated in A549 cell lines compared to normal lung epithelial cell BEAS-2B. MiR-132 overexpression obviously inhibited migration and invasion capacities in A549 cells while miR-132 down-regulation would enhance such capacities. Expression of EMT-related markers and TGFß1/Smad2 was higher in A549 cells transfected with miR-132 inhibitor compared with those transfected with miR-132 mimic. Moreover, expression of EMT-related markers and Smad2 was increased in NSCLC tissues compared to in the adjacent normal tissues and the reverse expression of miR-132 and Smad2 was observed. CONCLUSIONS: These results indicate that miR-132 may play a suppressive role in the metastasis of NSCLC cells by promoting EMT via TGFß1/Smad2 signaling pathway.

Overexpression of maelstrom promotes bladder urothelial carcinoma cell aggressiveness by epigenetically downregulating MTSS1 through DNMT3B.

We have recently identified and characterized a novel oncogene, maelstrom (MAEL) from 1q24, in the pathogenesis of hepatocellular carcinoma. In this study, MAEL was investigated for its oncogenic role in urothelial carcinoma of the bladder (UCB) tumorigenesis/aggressiveness and underlying molecular mechanisms. Here, we report that overexpression of MAEL in UCB is important in the acquisition of an aggressive and/or poor prognostic phenotype. In UCB cell lines, knockdown of MAEL by short hairpin RNA is sufficient to inhibit cell growth, invasiveness/metastasis and suppressed epithelial-mesenchymal transition (EMT), whereas ectopic overexpression of MAEL promoted cell growth, invasive and/or metastatic capacity and enhanced EMT both in vitro and in vivo. We further demonstrate that MAEL could induce UCB cell EMT by downregulating a critical downstream target, the metastasis suppressor 1 (MTSS1) gene, ultimately leading to an increased invasiveness of cancer cells. Notably, overexpression of MAEL in UCB cells substantially enhanced the enrichment of DNA methyltrans-ferase (DNMT)3B and histone deacetylase (HDAC)1/2 on the promoter of the MTSS1, and thereby epigenetically suppressing the MTSS1 transcription. Downregulation of MTSS1 by MAEL in UCB cells is partially dependent on DNMT3B. Furthermore, we identify that beside the gene amplification of MAEL, miR-186 is a key negative regulator of MAEL and downregulation of miR-186 is another important mechanism for MAEL overexpression in UCBs. These data suggest that overexpression of MAEL, caused by gene amplification and/or decreased miR-186, has a critical oncogenic role in UCB pathogenesis by downregulation of MTSS1, and MAEL could be used as a novel prognostic marker and/or effective therapeutic target for human UCB.

Cyclin I promotes cisplatin resistance via Cdk5 activation in cervical cancer.

OBJECTIVE: Cisplatin (cis-diamminedichloroplatinum II, CDDP) is one of the most effective chemotherapeutic agents and is widely used in the treatment of cervical cancer (CC), but cancer cell acquired resistance to this drug during the course of its treatment. The aim of this study was to investigate the role of cyclin I to cisplatin resistance in CC cell. PATIENTS AND METHODS: Cervical tumor specimens from 30 patients were recruited in this study. We analyzed the expression of cyclin I by real-time polymerase chain reaction (qRT-PCR), Western blotting examination of downstream effectors. Cell proliferation assay and xenograft experiments were performed for cisplatin cytotoxicity assay. Lentivirus-mediated and siRNA-mediated genes overexpression or knockdown were applied to investigate the role of cyclin I to cisplatin resistance in CC cell. RESULTS: We found that high level of cyclin I was associated with cisplatin resistance in CC. Here, we described that cyclin I protein becomes highly expressed in human CC patients resistant to cisplatin chemotherapy. Stable overexpressed cyclin I promotes Hela cell resistance to higher concentrations of cisplatin. In addition, upregulated level of cyclin I increased tumor cells growth in vitro and enhanced tumor resistance to cisplatin in vivo. The further mechanism investigated showed that cyclin I upregulated the expression of cyclin-dependent kinase 5 (Cdk5) promoting cisplatin resistance by preventing apoptosis in CC cell line. Consistently, the cyclin I overexpressed Hela cell lines produce increased sensitivity to cisplatin treatment through knockdown of Cdk5 protein with siRNA. CONCLUSIONS: These data suggest that a cyclin I-Cdk5 complex forms a critical antiapoptotic factor in the process of generating cisplatin resistance in cervical cancer.

Longikaurin A, a natural ent-kaurane, induces G2/M phase arrest via downregulation of Skp2 and apoptosis induction through ROS/JNK/c-Jun pathway in hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer, and is also highly resistant to conventional chemotherapy treatments. In this study, we report that Longikaurin A (LK-A), an ent-kaurane diterpenoid isolated from the plant Isodon ternifolius, induced cell cycle arrest and apoptosis in human HCC cell lines. LK-A also suppressed tumor growth in SMMC-7721 xenograft models, without inducing any notable major organ-related toxicity. LK-A treatment led to reduced expression of the proto-oncogene S phase kinase-associated protein 2 (Skp2) in SMMC-7721 cells. Lower Skp2 levels correlated with increased expression of p21 and p-cdc2 (Try15), and a corresponding decrease in protein levels of Cyclin B1 and cdc2. Overexpression of Skp2 significantly inhibited LK-A-induced cell cycle arrest in SMMC-7721 cells, suggesting that LK-A may target Skp2 to arrest cells at the G2/M phase. LK-A also induced reactive oxygen species (ROS) production and apoptosis in SMMC-7721 cells. LK-A induced phosphorylation of c-Jun N-terminal kinase (JNK), but not extracellular signal-regulated kinase and P38 MAP kinase. Treatment with, the JNK inhibitor SP600125 prevented LK-A-induced apoptosis in SMMC-7721 cells. Moreover, the antioxidant N-acetylcysteine prevented phosphorylation of both JNK and c-Jun. Taken together, these data indicate that LK-A induces cell cycle arrest and apoptosis in cancer cells by dampening Skp2 expression, and thereby activating the ROS/JNK/c-Jun signaling pathways. LK-A is therefore a potential lead compound for development of antitumor drugs targeting HCC.

EIF5A2 predicts outcome in localised invasive bladder cancer and promotes bladder cancer cell aggressiveness in vitro and in vivo.

BACKGROUND: EIF5A2, eukaryotic translation initiation factor 5A2, is associated with several human cancers. In this study, we investigated the role of EIF5A2 in the metastatic potential of localised invasive bladder cancer (BC) and its underlying molecular mechanisms were explored. METHODS: The expression pattern of EIF5A2 in localised invasive BC was determined by immunohistochemistry. In addition, the function of EIF5A2 in BC and its underlying mechanisms were elucidated with a series of in vitro and in vivo assays. RESULTS: Overexpression of EIF5A2 was an independent predictor for poor metastasis-free survival of localised invasive BC patients treated with radical cystectomy. Knockdown of EIF5A2 inhibited BC cell migratory and invasive capacities in vitro and metastatic potential in vivo and reversed epithelial-mesenchymal transition (EMT), whereas overexpression of EIF5A2 promoted BC cells motility and invasiveness in vitro and metastatic potential in vivo and induced EMT. In addition, we found that EIF5A2 might activate TGF-ß1 expression to induce EMT and drive aggressiveness in BC cells. EIF5A2 stabilized STAT3 and stimulated nuclear localisation of STAT3, which resulted in increasing enrichment of STAT3 onto TGF-ß1 promoter to enhance the transcription of TGF-ß1. CONCLUSIONS: EIF5A2 overexpression predicts tumour metastatic potential in patients with localised invasive BC treated with radical cystectomy. Furthermore, EIF5A2 elevated TGF-ß1 expression through STAT3 to induce EMT and promotes aggressiveness in BC.

AIB1 predicts bladder cancer outcome and promotes bladder cancer cell proliferation through AKT and E2F1.

BACKGROUND: We previously demonstrated that AIB1 overexpression is an independent molecular marker for shortened survival of bladder cancer (BC) patients. In this study, we characterised the role and molecular mechanisms of AIB1 in BC tumorigenicity. METHODS: AIB1 expression was measured by immunohistochemistry in non-muscle-invasive BC tissue and adjacent normal bladder tissue. In addition, the tumorigenicity of AIB1 was assessed with in vitro and in vivo functional assays. RESULTS: Overexpression of AIB1 was observed in tissues from 46 out of 146 patients with non-muscle-invasive BC and was an independent predictor for poor progression-free survival. Lentivirus-mediated AIB1 knockdown inhibited cell proliferation both in vitro and in vivo, whereas AIB1 overexpression promoted cell proliferation in vitro. The growth-inhibitory effect induced by AIB1 knockdown was mediated by G1 arrest, which was caused by reduced expression of key cell-cycle regulatory proteins through the AKT pathway and E2F1. CONCLUSION: Our results suggest that AIB1 promotes BC cell proliferation through the AKT pathway and E2F1. Furthermore, AIB1 overexpression predicts tumour progression in patients with non-muscle-invasive BC.

N-acetylcysteine attenuates subcutaneous administration of bleomycin-induced skin fibrosis and oxidative stress in a mouse model of scleroderma.

BACKGROUND: Several lines of evidence suggest that the generation of reactive oxygen species (ROS) is of major importance in the pathogenesis of scleroderma, and thus antioxidant therapy may be useful for patients with an impaired oxidative defence mechanism. AIM: To examine the effect of N-acetylcysteine (NAC) on skin fibrosis and oxidative stress in a bleomycin (BLM)-induced mouse model of scleroderma. METHODS: We used this mouse model to evaluate the effect of NAC on skin fibrosis and oxidative stress. Skin fibrosis was evaluated by histopathological examination and hydroxyproline content. To measure lipid peroxidation, we used a thiobarbituric acid-reactive species, malondialdehyde (MDA). Oxidative protein damage (carbonyl content) and the activities of catalase (CAT) and superoxide dismutase (SOD) were determined to evaluate oxidative stress in the skin tissue. RESULTS: Treatment with NAC attenuated the skin fibrosis induced by BLM, significantly reducing the MDA and protein carbonyl content in these mice. SOD activity in BLM-only mice and BLM plus NAC-treated mice was increased compared with control mice. However, there was no significant difference in skin SOD activity of mice treated with both BLM and NAC compared with those treated with BLM only. In addition, CAT activity was not altered in the BLM plus NAC mice. CONCLUSIONS: NAC treatment attenuates skin fibrosis in a BLM-induced mouse model of scleroderma, and this is associated with diminished oxidative stress. The results suggest that NAC may be a potential therapeutic agent for patients with scleroderma.

Inactivation of the bed nucleus of the stria terminalis suppresses the innate fear responses of rats induced by the odor of cat urine.

In this study, we investigated whether two brain regions, the bed nucleus of the stria terminalis (BNST) and the basolateral amygdala (BLA), affected male rats' (Rattus norvigicus) ability to innately discriminate between a predator odor (cat urine) and female rat urine. Muscimol, a GABAa receptor agonist, was bilaterally microinjected into either the BNST or BLA of rats through implanted stainless-steel guide cannulas to temporarily inactivate these brain nuclei. The behavioral responses of the treated rats to female rat urine and cat urine were then tested in an experimental arena. Compared to a saline infusion control, the injection of muscimol into the BNST strongly reversed the innate aversion of rats to cat urine but the injection of muscimol into the BLA had no effect. Furthermore, intra-BNST infusion of muscimol caused rats to be equally attracted to urine from cats and female rats but intra-BLA infusion did not stop rats manifesting fear on exposure to cat urine and exploratory behavior on exposure to female rat urine. We conclude that the BNST plays a more crucial role in modulating innate fear responses in rats than the BLA.