Mining biomarkers from routine laboratory tests in clinical records associated with air pollution health risk assessment.

Clinical laboratory in hospital can produce amounts of health data every day. The purpose of this study was to mine biomarkers from clinical laboratory big data associated with the air pollution health risk assessment using clinical records. 13, 045, 629 clinical records of all 27 routine laboratory tests in Changsha Central Hospital, including ALB, TBIL, ALT, DBIL, AST, TP, UREA, UA, CREA, GLU, CK, CKMB, LDL-C, TG, TC, HDL-C, CRP, WBC, Na, K, Ca, Cl, APTT, PT, FIB, TT, RBC and those daily air pollutants concentration monitoring data of Changsha, including PM2.5, PM10, SO2, NO2, CO, and O3 from 2014 to 2016, were retrieved. The moving average method was used to the biological reference interval was established. The tests results were converted into daily abnormal rate. After data cleaning, GAM statistical model construction and data analysis, a concentration-response relationship between air pollutants and daily abnormal rate of routine laboratory tests was observed. Our study found that PM2.5 had a stable association with TP (lag07), ALB (lag07), ALT (lag07), AST (lag07), TBIL (lag07), DBIL (lag07), UREA (lag07), CREA (lag07), UA (lag07), CK (lag 06), GLU (lag07), WBC (lag07), Cl (lag07) and Ca (lag07), (P < 0.05); O3 had a stable association with AST (lag01), CKMB (lag06), TG (lag07), TC (lag05), HDL-C (lag07), K (lag05) and RBC (lag07) (P < 0.05); CO had a stable association with UREA (lag07), Na (lag7) and PT (lag07) (P < 0.05); SO2 had a stable association with TP (lag07) and LDL-C (lag0) (P < 0.05); NO2 had a stable association with APTT (lag7) (P < 0.05). These results showed that different air pollutants affected different routine laboratory tests and presented different pedigrees. Therefore, biomarkers mined from routine laboratory tests may potentially be used to low-cost assess the health risks associated with air pollutants.

Inhibition of LONP1 protects against erastin-induced ferroptosis in Pancreatic ductal adenocarcinoma PANC1 cells.

Ferroptosis is identified as a regulated cell death mediated by iron accumulation and lipid peroxidation. The disturbances of mitochondrial morphology and function have been shown in this process. Mitochondrial Lon peptidase 1 (LONP1) is one of the main multi-function enzymes in regulating the mitochondrial function and cytological stability. To evaluate whether LONP1 take a role in ferroptosis, we applied erastin to initiate the ferroptosis in human pancreatic ductal adenocarcinoma (PDAC) cells. Here we show that erastin triggers cell death in both of oncogenic RAS mutant PANC1 cells and wild KRAS BxPC3 cells and the expression of LONP1 was up-regulated in this process. Gene inhibition of LONP1 only negatively regulates erastin-induced cell death and the alterations of molecular indicators in PANC1 cells. Furthermore, we show that inhibition of LONP1 activates the Nrf2/Keap1 signal pathway and up-regulates the expression of GPX4, a key peroxidase in regulating ferroptosis. Together, our results uncover a previously unappreciated mechanism coupling LONP1 to ferroptosis.

APLNR is involved in ATRA-induced growth inhibition of nasopharyngeal carcinoma and may suppress EMT through PI3K-Akt-mTOR signaling.

The apelin receptor (APLNR) is a GPCR involved in many pathophysiological processes; however, the correlation between APLNR expression and nasopharyngeal carcinoma (NPC) has not been reported. In this study, we used cDNA microarray data to determine APLNR expression levels in NPC tissues. We found that APLNR expression was reduced in NPC tissues compared with noncancerous nasopharyngeal epithelial tissues. Subsequently, a large-scale sample of 1015 tissues was used to validate this discovery and explore the relationship between APLNR expression and prognosis of NPC. Expression levels of APLNR in NPC tissues were indeed down-regulated. Furthermore, positive expression of APLNR in NPC predicted a better prognosis (disease-free survival: P = 0.001; overall survival: P < 0.001). Moreover, ingenuity pathway analysis revealed that an indirect interaction existed between APLNR and retinoic acid (RA) in the cancer regulatory network. Consistently, after treatment with all-trans-RA (ATRA), we found that APLNR was significantly up-regulated in NPC cell lines (5-8F and HNE1), and proliferation of NPC cells was inhibited. Cell cycle arrest occurred in the G0/G1 phase. In contrast, knockdown of APLNR diminished ATRA-induced growth inhibition of NPC cells. In addition, we surprisingly found that APLNR also played an important role in migration and invasion of NPC. Wound-healing and Transwell assays revealed that APLNR overexpression led to reduced migratory and invasive properties in 2 NPC cell lines. Western blot results revealed that hallmarks of epithelial-mesenchymal transition (EMT) were altered as well, suggesting that APLNR was capable of inhibiting EMT in NPC cells. Our study further demonstrated that low expression of APLNR promoted EMT in NPC cells by activating the PI3K-protein kinase B-mammalian target of rapamycin signaling pathway. Taken together, our data suggest that APLNR could potentially predict prognosis for patients with NPC and inhibit proliferation, migration, invasion, and EMT in nasopharyngeal cancer cells.-Liu, Y., Liu, Q., Chen, S., Liu, Y., Huang, Y., Chen, P., Li, X., Gao, G., Xu, K., Fan, S., Zeng, Z., Xiong, W., Tan, M., Li, G., Zhang, W. APLNR is involved in ATRA-induced growth inhibition of nasopharyngeal carcinoma and may suppress EMT through PI3K-Akt-mTOR signaling.

Inhibin B suppresses anoikis resistance and migration through the transforming growth factor-ß signaling pathway in nasopharyngeal carcinoma.

Inhibin B (INHBB), a heterodimer of a common α-subunit and a ßB-subunit, is a glycoprotein belonging to the transforming growth factor-ß (TGF-ß) family. In this study, we observed INHBB expression was reduced in nasopharyngeal carcinoma (NPC) tissues compared to non-tumor nasopharyngeal epithelium tissues, and INHBB was associated with lymph node metastasis, stage of disease, and clinical progress. Positive expression of INHBB in NPC predicted a better prognosis (overall survival, P = 0.038). However, the molecular mechanisms of INHBB have not been addressed in NPC. We induced anoikis-resistant cells in NPC cell lines under anchorage-independent conditions, then found epithelial-mesenchymal transition markers changed, cell apoptosis decreased, cell cycle was modified, and invasion strengthened in anoikis-resistant NPC cells. These anoikis-resistant NPC cells showed decreased expression of INHBB compared with adhesion cells. Furthermore, INHBB was found to influence the above-mentioned changes. In the anoikis-resistant NPC cells with INHBB overexpression, apoptotic cells increased, S phase cells weakened, vimentin, matrix metallopeptidase-9, and vascular endothelial growth factor A expression were downregulated, and E-cadherin expression was upregulated, and vice versa in knockdown of INHBB (INHBB shRNA) anoikis-resistant NPC cells. Diminished INHBB expression could activate the TGF-ß pathway to phosphorylate Smad2/3 and form complexes in the nucleus, which resulted in the above changes. Thus, our results revealed for the first time that INHBB could suppress anoikis resistance and migration of NPC cells by the TGF-ß signaling pathway, decrease p53 overexpression, and could serve as a potential biomarker for NPC metastasis and prognosis as well as a therapeutic application.

[Optimization of sperm alkaline single-cell gel electrophoresis].

OBJECTIVE: To investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures. METHODS: Using alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method. RESULTS: Three agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05). CONCLUSION: The optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

[Improvement of immunoprecipitation/immunoblotting of endogenous protein by TrueBlot antibody].

OBJECTIVE: To improve the immunoblotting of immunoprecipitated proteins and decrease the interference of immunoprecipitation antibody in the interaction of endogenous proteins. METHODS: Transient transfect cells with fusion protein expression vector containing the targeted S5b gene and the FLAG tag, the transfected cells or untransfected cells were harvested to study the exogenous or endogenous protein interaction. The total cell lysate was immunoprecipitated by specific antibody. Then the eluted immunocomplex was separated by SDS-PAGE, and the TrueBlot antibody or conventional antibody was used as the secondary antibody for immunoblotting detection of S5b and its partner (Rpt1 and Rpt2). RESULTS: Clear immunoblotting bands for S5b, Rpt1 and Rpt2 were obtained. CONCLUSION: TrueBlot antibody prefers the immunoblot antibody to immunoprecipitation antibody, and decreases the interruption of immunoprecipitation antibody to display clear protein band.

Assessment of anti-recombination and double-strand break-induced gene conversion in human cells by a chromosomal reporter.

Gene conversion is one of the frequent end results of homologous recombination, and it often underlies the inactivation of tumor suppressor genes in cancer cells. Here, we have developed an integrated assay system that allows simultaneous examination of double-strand break (DSB)-induced gene conversion events at the site of a DSB (proximal region) and at a surrounding region ~1 kb away from the break (distal region). Utilizing this assay system, we find that gene conversion events at the proximal and distal regions are relatively independent of one another. The results also indicate that synthesis-dependent strand annealing (SDSA) plays a major role in DSB-induced gene conversion. In addition, our current study has demonstrated that hMLH1 plays an essential role in anti-recombination and gene conversion. Specifically, the anti-recombination activity of hMLH1 is partially dependent on its interaction with hMRE11. Our data suggests that the role of hMLH1 and hMRE11 in the process of gene conversion is complex, and these proteins play different roles in DSB-induced proximal and distal gene conversions. In particular, the involvement of hMLH1 and hMRE11 in the distal gene conversion requires both hMSH2 and heteroduplex formation.

[Rapid detection of microalbuminuria with high-performance liquid chromatography].

OBJECTIVE: To develop a rapid detection method for microalbuminuria with size-exclusion high performance liquid chromatography. METHODS: With a new mobile phase, samples were injected onto an Agilent Zorbax GF-250 size exclusion chromatographic column. The method was evaluated and urine albumin of 56 diabetic patients was analyzed. RESULTS: The mobile phase containing 0.1% formic acid and acetonitrile, with 20 µL sample size, 1 mL/min flow rate, 205 nm detection wavelength. The retention time of albumin both in human serum and urine was 1.7 min. The linear range was 5-2000 mg/L. The lower limit of measurement was 2 mg/L. The intra-assay coefficient of variation and the inter-assay coefficient of variation were 3.98% and 4.05% (20 mg/L), 3.55% and 3.60% (200 mg/L), 4.65% and 4.74% (2000 mg/L), respectively. Recovery rates were 95.3%, 98.1%, and 97.2%. Microalbuminuria was detected in 30 samples by high performance liquid chromatography and 15 samples by immunoturbidimetry from 56 patients with diabetic mellitus. CONCLUSION: A fast and high sensitivity method, namely size-exclusion high performance liquid chromatography, with mobile phase containing 0.1% formic acid and acetonitrile has been established to analyze microalbuminuria, which can detect more microalbuminuria than other methods and is suitable for clinical routine measurement.

Ku70 affects the frequency of chromosome translocation in human lymphocytes after radiation and T-cell acute lymphoblastic leukemia.

BACKGROUND: As one of the most common chromosomal causes, chromosome translocation leads to T-cell acute lymphoblastic leukemia (T-ALL). Ku70 is one of the key factors of error-prone DNA repair and it may end in translocation. So far, the direct correlation between Ku70 and translocation has not been assessed. This study aimed to investigate the association between Ku70 and translocation in human lymphocytes after radiation and T-ALL. METHODS: Peripheral blood lymphocytes (PBLs) from volunteers and human lymphocyte cell line AHH-1 were irradiated with X-rays to form the chromosome translocations. Phytohemagglutinin (PHA) was used to stimulate lymphocytes. The frequency of translocation was detected by fluorescence in situ hybridization (FISH). Meanwhile, the expression of Ku70 was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. Furthermore, Ku70 interference, overexpression and chemical inhibition were used in AHH-1 cell lines to confirm the correlation. Finally, the expression of Ku70 in T-ALL samples with or without translocation was detected. RESULTS: The expression of Ku70 and frequencies of translocation were both significantly increased in PBLs after being irradiated by X-rays, and a positive correlation between the expression (both mRNA and protein level) of Ku70 and the frequency of translocation was detected (r = 0.4877, P = 0.004; r = 0.3038, P = 0.0358 respectively). Moreover, Ku70 interference decreased the frequency of translocations, while the frequency of translocations was not significantly affected after Ku70 overexpression. The expression of Ku70 and frequencies of translocation were both significantly increased in cells after irradiation, combined with chemical inhibition (P < 0.01). The protein level and mRNA level of Ku70 in T-ALL with translocation were obviously higher than T-ALL with normal karyotype (P = 0.009, P = 0.049 respectively). CONCLUSIONS: Ku70 is closely associated with the frequency of chromosome translocation in human lymphocytes after radiation and T-ALL. Ku70 might be a radiation damage biomarker and a potential tumor therapy target.

Polymorphisms Within DNA Double-Strand Breaks Repair-Related Genes Contribute to Structural Chromosome Abnormality in Recurrent Pregnancy Loss.

Background: Structural chromosome abnormality (SCA) is an important cause of human diseases, including recurrent pregnancy loss (RPL). DNA double-strand breaks (DSBs) repair-related genes play critical roles in SCA. The present study aims to investigate the potential contribution of DSBs repair-related gene polymorphisms to SCA. Methods: Fifty-four affected RPL individuals with SCA, 88 affected RPL individuals without SCA, and 84 controls were analyzed. Targeted whole-exome sequencing (WES) was used for screening single nucleotide polymorphisms in six DSBs repair-related genes (EP300, XRCC6, LIG4, XRCC4, PRKDC, and DCLRE1C), and validation was performed by Sanger sequencing. Finally, we detected the frequency of radiation-induced chromosome translocations in no SCA samples with significant polymorphisms by fluorescence in situ hybridization (FISH). Results: A total of 35 polymorphisms have been identified and confirmed. Frequencies of EP300 rs20551, XRCC6 rs132788, and LIG4 rs1805388 were significantly different between SCA RPL and no SCA RPL (p = 0.030, 0.031, and 0.040 respectively). Frequencies of those three gene polymorphisms between SCA RPL and controls also were significantly different (p = 0.017, 0.028, and 0.029 respectively). Moreover, the frequency of the G allele at rs20551 locus, the T allele at rs132788 locus and the A allele at rs1805388 locus was significantly higher in SCA RPL than no SCA RPL (OR = 3.227, p = 0.005; OR = 1.978, p = 0.008 and OR = 1.769, p = 0.036 respectively) and controls (OR = 7.130, p = 0.000; OR = 2.157, p = 0.004; OR = 2.397, p = 0.003 respectively). Additionally, the frequency of radiation-induced translocation in no SCA samples with rs20551, rs132788 or rs1805388 was significantly higher compared with the wild type samples (p = 0.015, 0.012, and 0.007 respectively). Conclusion: Our results suggest that rs20551, rs132788, and rs1805388 might be associated with the risk of SCA. Larger scales of genetic variations studies and functional experiments are necessary to further confirm these findings.

Association of hMSH5 C85T polymorphism with radiation sensitivity of testicular cell lines GC-1, GC-2, TM3, and TM4.

BACKGROUND: The hMSH5 C85T polymorphism, which encodes hMSH5 P29S, is associated with individual differences in spermatogenic abnormalities caused by ionizing radiation (IR), but the molecular mechanisms remain unclear. OBJECTIVES: This manuscript aims to explore the role of hMSH5 C85T polymorphism in IR-induced individual differences in spermatogenic abnormalities. MATERIAL AND METHODS: We transfected pcDNA-hMSH5P29S vector into mouse spermatogonia GC-1, mouse spermatocytes GC-2, mouse testicular mesenchymal cells TM3, and mouse testicular support cells TM4. After radiation, we evaluated cell survival with colony formation assay, apoptosis with TUNEL assay and caspase-3 activity assay, DNA damage with comet assay and an in vivo NHEJ activity assay. RESULTS: Results showed that only spermatocytes GC-2 transfected with pcDNA-hMSH5P29S vector had significant differences in IR-induced cell survival and apoptosis when compared to that transfected with pcDNA empty vector and pcDNA-wild-hMSH5 vector, while there was no statistical difference in GC-1, TM3, and TM4. In addition, comet assay showed that the DNA damage of GC-2 transfected with pcDNA-hMSH5P29S vector increased significantly compared to that transfected with pcDNA empty vector and pcDNA-wild-hMSH5 vector after IR. And in vivo NHEJ activity assay showed that the NHEJ activity of GC-2 transfected with pcDNA-hMSH5P29S vector was statistically higher than that transfected with pcDNA empty vector and pcDNA-wild-hMSH5 vector. CONCLUSION: Our study indicates that the hMSH5 C85T polymorphism leads to an abnormal increase in apoptosis and lessen the control on error-prone NHEJ of spermatocyte GC-2, thereby altering the difference of radiation sensitivity of spermatogenesis.

Comprehensive analysis and identification of key genes and signaling pathways in the occurrence and metastasis of cutaneous melanoma.

BACKGROUND: Melanoma is a malignant tumor of melanocytes, and the incidence has increased faster than any other cancer over the past half century. Most primary melanoma can be cured by local excision, but metastatic melanoma has a poor prognosis. Cutaneous melanoma (CM) is prone to metastasis, so the research on the mechanism of melanoma occurrence and metastasis will be beneficial to diagnose early, improve treatment, and prolong life survival. In this study, we compared the gene expression of normal skin (N), primary cutaneous melanoma (PM) and metastatic cutaneous melanoma (MM) in the Gene Expression Omnibus (GEO) database. Then we identified the key genes and molecular pathways that may be involved in the development and metastasis of cutaneous melanoma, thus to discover potential markers or therapeutic targets. METHODS: Three gene expression profiles (GSE7553, GSE15605 and GSE46517) were downloaded from the GEO database, which contained 225 tissue samples. R software identified the differentially expressed genes (DEGs) between pairs of N, PM and MM samples in the three sets of data. Subsequently, we analyzed the gene ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of the DEGs, and constructed a protein-protein interaction (PPI) network. MCODE was used to seek the most important modules in PPI network, and then the GO function and KEGG pathway of them were analyzed. Finally, the hub genes were calculated by the cytoHubba in Cytoscape software. The Cancer Genome Atlas (TCGA) data were analyzed using UALCAN and GEPIA to validate the hub genes and analyze the prognosis of patients. RESULTS: A total of 134, 317 and 147 DEGs were identified between N, PM and MM in pair. GO functions and KEGG pathways analysis results showed that the upregulated DEGs mainly concentrated in cell division, spindle microtubule, protein kinase activity and the pathway of transcriptional misregulation in cancer. The downregulated DEGs occurred in epidermis development, extracellular exosome, structural molecule activity, metabolic pathways and p53 signaling pathway. The PPI network obtained the most important module, whose GO function and KEGG pathway were enriched in oxidoreductase activity, cell division, cell exosomes, protein binding, structural molecule activity, and metabolic pathways. 14, 18 and 18 DEGs were identified respectively as the hub genes between N, PM and MM, and TCGA data confirmed the expression differences of hub genes. In addition, the overall survival curve of hub genes showed that the differences in these genes may lead to a significant decrease in overall survival of melanoma patients. CONCLUSIONS: In this study, several hub genes were found from normal skin, primary melanoma and metastatic melanoma samples. These hub genes may play an important role in the production, invasion, recurrence or death of CM, and may provide new ideas and potential targets for its diagnosis or treatment.

Association between fine particle exposure and common test items in clinical laboratory: A time-series analysis in Changsha, China.

Most studies on the health effects of PM2.5 (fine particulate matter with diameter smaller than 2.5 µm) use indirect indicators, such as mortality and number of hospital visits. Recent research shows that biomarkers can also be used to evaluate the health effects of PM2.5; however, these biomarkers are not very common. Clinical laboratories can provide a significant amount of test data that have been proven to have important diagnostic value. Therefore, we use big data analysis methods to find the associations between clinical laboratory common test items and PM2.5 exposure. Data related to air pollution and meteorological information between 2014 and 2016 were obtained from the China National Environmental Monitoring Centre and the China National Meteorological Information Center. Additionally, data of 27 common test items from the same period were collected from Changsha Central Hospital. Primary analyses included a generalized additive model to analyze the associations between PM2.5 concentration and common test items; the model was adjusted for time trends, weather conditions (temperature and humidity), and days of the week. Furthermore, we adjusted the effects of other air pollutants, such as PM10, SO2, NO2, CO, and O3. 17 items such as TP, ALB, ALT, AST, TBIL, DBIL, UREA, CREA, UA, GLU, LDL, WBC, K, Cl, Ca, TT, and FIB were significantly positively associated with PM2.5 concentration (P< 0.05) and have concentration-response relationship. After adjusting the effect of PM10+SO2+NO2+CO+O3, TP, ALB, ALT, AST, TBIL, DBIL, UREA, CREA, UA, GLU, WBC, Cl, and Ca were still significantly associated with PM2.5 concentration (P< 0.05). This current study suggested that clinical laboratory common test items may be used to assess and predict the health effects of PM2.5 on the population.

Jujuboside B promotes the death of acute leukemia cell in a RIPK1/RIPK3/MLKL pathway-dependent manner.

Initiation of necroptosis has been considered as a promising strategy for anticancer therapies, especially for eradicating apoptosis-resistant malignant cells. Jujubisode B is a natural saponins extracted from the seeds of Zizyphi Spinosi Semen, and possesses multiple pharmacological activities, including antianxiety, anti-inflammation, antiplatelet aggregation and induction of apoptosis. This study aims to explore the effect of jujuboside B on acute leukemic cells and the underlying mechanisms. Our results showed that jujuboside B inhibited leukemia cell growth in a dose-dependent manner and attenuated the clonogenic ability of U937 cells, concomitant with activation of RIPK1/RIPK3/MLKL pathway; these phenomena were evidently blocked by necroptosis inhibitor (Nec-1). With the help of Molecular Operating Environment (MOE) program, we identified that RIPK1, RIPK3 and MLKL are potential targets of jujuboside B. To the best of our knowledge, this is the first study to provide evidence that jujuboside B possesses antileukemic activity via a mechanism involving activation of RIPK1/RIPK3/MLKL pathway.

[Effect of fluorouracil combined with FK228 on the proliferation, apoptosis and Fas mRNA level in HepG2 hepatoma cell lines].

OBJECTIVE: To explore the effect of fluorouracil (FU) combined with epigenetic drug FK228 (depsipeptide FR901228) on the proliferation, apoptosis and Fas mRNA level in HepG2 hepatoma cell lines. METHODS: There were 4 groups in this experiment: an untreated group, an FK228 treated group, a FU treated group,and a FU combined with FK228 group.Tetrazolium salt colorimetry (MTT) assay was used to assess the cell inhibition rate. Q value of Kingos formula and multi-factor analysis of variance (ANOVA ) were used to judge the combination treatment effect,and flow cytometry was used to detect the apoptosis rate. Fas mRNA level was analyzed by RT-PCR. RESULTS: FK228 or FU would inhibit the growth of HepG2 cells in a concentration-dependent and time-dependent manner. Cell inhibition rates of HepG2 were significantly enhanced in the FU combined with FK228 group, compared with that in the FU treated group alone (P<0.05). Both Q values were more than 1, the 2 drug combinations showed interaction,and FU combined with FK228 had synergistic effect.Compared with the FK228 treated group and the FU treated group, apoptosis rate of HepG2 cells was significantly increased (P<0.05), and the Fas mRNA level was up-regulated in HepG2 cells in FU combined FK228 group (P<0.05). CONCLUSION: Combination of FK228 and FU can enhance the proliferation inhibition and apoptosis induction of FU in hepatoma cell lines, up-regulate the Fas mRNA level, and increase the sensitivity of hepatoma cell lines to FU.

SIRT6, a novel direct transcriptional target of FoxO3a, mediates colon cancer therapy.

SIRT6, NAD+-dependent deacetylase sirtuin 6, has recently shown to suppress tumor growth in several types of cancer. Colon cancer is a challenging carcinoma associated with high morbidity and death. However, whether SIRT6 play a direct role in colon tumorigenesis and the underlying mechanism are not understood. Methods: To investigate the role of SIRT6 in colon cancer, we firstly analyzed the specimens from 50 colorectal cancer (CRC) patients. We generated shSIRT6 LoVo cells and xenograft mouse to reveal the essential role of SIRT6 in cell apoptosis and tumor growth. To explore the underlying mechanism of SIRT6 regulation, we performed FRET and real-time fluorescence imaging in living cells, real-time PCR, immunoprecipitaion, immunohistochemistry, flow cytometry and luciferase reporter assay. Results: The expression level of SIRT6 in patients' specimens is lower than that of normal controls, and patients with higher SIRT6 level have a better prognosis. Here, we identified that transcriptional factor FoxO3a is a direct up-stream of SIRT6 and positively regulated SIRT6 expression, which in turn, promotes apoptosis by activating Bax and mitochondrial pathway. Functional studies reveal that Akt inactivation increases FoxO3a activity and augment its binding to SIRT6 promoter, leading to elevated SIRT6 expression. Knocking down SIRT6 abolished apoptotic responses and conferred resistance to the treatment of BKM120. Combinational therapies with conventional drugs showed synergistic chemosensitization, which was SIRT6-dependent both in vitro and in vivo. Conclusion: The results uncover SIRT6 as a new potential biomarker for colon cancer, and its unappreciated mechanism about transcription and expression via Akt/FoxO3a pathway.

Identification of key genes and multiple molecular pathways of metastatic process in prostate cancer.

BACKGROUND: Cancer metastasis is well known as the most adverse outcome and the major cause of mortality in cancer patients, including prostate cancer (PCa). There are no credible predictors, to this day, that can reflect the metastatic ability of localized PCa. In the present study, we firstly identified the differentially expressed genes (DEGs) and molecular pathways involved in the metastaic process of PCa by comparing gene expressions of metastaic PCa with localized PCa directly, with the purpose of identifying potential markers or therapeutic targets. METHODS: The gene expression profiles (GSE6919 and GSE32269) were downloaded from the Gene Expression Omnibus database, which contained 141 tissue samples, including 87 primary localized PCa samples and 54 metastaic PCa samples. After data processing, DEGs were identified by R language using the Student's t-test adjusted via the Beniamini-Hochberg method. Subsequently, the gene ontology functional and pathway enrichment analyses of DEGs were performed and the protein-protein interaction network was constructed. Hub genes were identified using the plug-in cytoHubba in Cytoscape software by MCC and degree. Furthermore, validation and prognostic significance analysis of the hub genes were performed by UALCAN and gene expression profiling interactive analysis (GEPIA). RESULTS: A total of 90 DEGs were identified between localized and metastaic PCa, which consisted of 47 upregulated and 43 downregulated genes. The enriched functions and pathways of the DEGs include catabolic process, cell cycle, response to steroid hormone, extracellular matrix (ECM)-receptor interaction and vascular smooth muscle contraction. A total of 10 genes were identified as hub genes and biological process analysis of hub genes showed that cell cycle phase, cell division, and mitotic cell cycle process were mainly enriched. The expression of hub genes were confirmed in metastaic PCa when compared with localized PCa tissues by The Cancer Genome Atlas database. Moreover, the disease-free survival analysis of hub genes revealed that these genes may play an important role in invasion, progression or recurrence. Therefore, these hub genes might be the key genes contributed to tumor progression or metastasis in PCa and provide candidate therapeutic targets for PCa. CONCLUSIONS: The present study identified some DEGs between localized and metastaic PCa tissue samples. These key genes might be potential therapeutic targets and biomarkers for the metastaic process of PCa.

Long-term high air pollution exposure induced metabolic adaptations in traffic policemen.

OBJECTIVE: To assess the adverse physiological changes induced by long-term exposure to PM2.5. METHODS: Totally 183 traffic policemen and 88 office policemen as the control group, were enrolled in this study. The concentrations of PM2.5 in both the working places of traffic and office policemen were obtained. Detailed personal questionnaires and conventional laboratory tests including hematology, fasting blood glucose, blood lipids, liver, kidney, immunity and tumor-related markers were conducted on all participants of this study. RESULTS: A dose-response relationship between the FBG, HDL-c and CEA values and the PM2.5 exposure duration was observed. Multivariate analysis confirmed that one hour on duty outdoor per day for one year was associated with an increase in FBG of 0.005% (95% CI: 0.0004% to 0.009%), CEA of 0.012% (95% CI: 0.006% to 0.017%), and a decrease in HDL-C of 0.001% (95% CI: 0.00034% to 0.002%). CONCLUSION: Long-term high air pollution exposure may lead to metabolism adaptation and it is likely involved in the development of cardiovascular disease and diabetes mellitus.

Long-term exposure to high air pollution induces cumulative DNA damages in traffic policemen.

The specific effects of long-term exposure to high air pollution on human health and biological remain unclear. To explore the adverse health effects as well as biological mechanisms and biomarkers for durative exposure to air pollution, 183 traffic policemen and 88 office policemen were enrolled in this study. The concentration of PM2.5 in both the traffic and office policemen's working environments were obtained. Detailed personal questionnaires were completed and levels of inflammation, oxidative stress and DNA damage markers of all participants were analyzed in this study. The average PM2.5 concentration of the intersections of main roads and the offices of control group were 132.4±48.9µg/m3 and 50.80±38.6µg/m3, respectively. The traffic policemen, who stably exposed to at least 2 times higher PM2.5 in their work area as compared with the control group, have a median average duration of 7.00years, and average cumulative intersection duty time reached 8030h. No statistically significant differences in the levels of inflammation markers were observed between the traffic and office policemen. However, the DNA damage markers in traffic policemen shared significant positive correlation with cumulative intersection duty time and higher than those in the office policemen. Multiple linear regression analysis demonstrated that the increase of cumulative intersection duty time by 1h per day for one year was associated with the increase in 8-hydroxy-20-deoxyguanosine of 0.329% (95% CI: 0.249% to 0.409%), tail DNA of 0.051% (95% CI: 0.041% to 0.061%), micronucleus frequency of 0.036‰ (95% CI: 0.03‰ to 0.043‰), and a decrease in glutathione of 0.482% (95% CI: -0.652% to -0.313%). These findings suggest that long-term exposure to high air pollution could induce cumulative DNA damages, supporting the hypothesis that durative exposure to air pollution is associated with an increased risk of cancer.