Mechanism and bioinformatics analysis of the effect of berberine-enhanced fluconazole against drug-resistant Candida albicans.

Biofilms produced by Candida albicans present a challenge in treatment with antifungal drug. Enhancing the sensitivity to fluconazole (FLC) is a reasonable method for treating FLC-resistant species. Moreover, several lines of evidence have demonstrated that berberine (BBR) can have antimicrobial effects. The aim of this study was to clarify the underlying mechanism of these effects. We conducted a comparative study of the inhibition of FLC-resistant strain growth by FLC treatment alone, BBR treatment alone, and the synergistic effect of combined FLC and BBR treatment. Twenty-four isolated strains showed distinct biofilm formation capabilities. The antifungal effect of combined FLC and BBR treatment in terms of the growth and biofilm formation of Candida albicans species was determined via checkerboard, time-kill, and fluorescence microscopy assays. The synergistic effect of BBR and FLC downregulated the expression of the efflux pump genes CDR1 and MDR, the hyphal gene HWP1, and the adhesion gene ALS3; however, the gene expression of the transcriptional repressor TUP1 was upregulated following treatment with this drug combination. Furthermore, the addition of BBR led to a marked reduction in cell surface hydrophobicity. To identify resistance-related genes and virulence factors through genome-wide sequencing analysis, we investigated the inhibition of related resistance gene expression by the combination of BBR and FLC, as well as the associated signaling pathways and metabolic pathways. The KEGG metabolic map showed that the metabolic genes in this strain are mainly involved in amino acid and carbon metabolism. The metabolic pathway map showed that several ergosterol (ERG) genes were involved in the synthesis of cell membrane sterols, which may be related to drug resistance. In this study, BBR + FLC combination treatment upregulated the expression of the ERG1, ERG3, ERG4, ERG5, ERG24, and ERG25 genes and downregulated the expression of the ERG6 and ERG9 genes compared with fluconazole treatment alone (p < 0.05).

Tumor-derived miRNAs as tumor microenvironment regulators for synergistic therapeutic options.

MicroRNAs (miRNAs) are a class of non-coding RNAs that perform post-transcriptional gene regulation. This review focuses on the role of tumor cell-derived miRNAs in the regulation of the tumor microenvironment (TME) via receptor cell recoding, including angiogenesis, expression of immunosuppressive molecules, formation of radiation resistance, and chemoresistance. Furthermore, we discuss the potential of these molecules as adjuvant therapies in combination with chemotherapy, radiotherapy, or immunotherapy, as well as their advantages as efficacy predictors for personalized therapy. MiRNA-based therapeutic agents for tumors are currently in clinical trials, and while challenges remain, additional research on tumor-derived miRNAs is warranted, which may provide significant clinical benefits to cancer patients.

The aberrant overexpression of hsa_circ_0078607 in colorectal cancer tissues and serum and its clinical significance / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究hsa_circ_0078607在结直肠癌组织和患者血清中的表达水平及其与结直肠癌患者临床病理特征的关系，评价其能否作为结直肠癌潜在的分子诊断标志物及治疗靶标。方法：收集2018年6月至2022年1月于柳州市人民医院胃肠外科接受结直肠癌切除手术患者的58对癌及癌旁组织标本，收集2020年1月至2022年12月于柳州市人民医院初次确诊的结直肠癌患者、结直肠息肉患者及健康人体检血清共152例；从结直肠癌差异表达circRNA谱中挑选特异性高表达的hsa_circ_0078607作为候选标志物，采用qPCR法检测其在结直肠癌细胞、组织、患者血清及结直肠息肉患者血清中的相对表达量，分析其与临床病理特征的关系。采用ROC曲线评估hsa_circ_0078607对结直肠癌及结直肠息肉的诊断价值。通过Circular RNA Interactome数据库预测与hsa_circ_0078607结合的miRNA，并用Cytoscape 3.9.1软件构建circRNA-miRNA-mRNA调控网络，同时通过GO/KEGG富集分析进一步了解其功能。结果：与癌旁组织或健康人血清相比，hsa_circ_0078607在结直肠癌细胞、组织和血清及息肉患者血清中呈高表达（P<0.001），其中有52例（89.7%）患者癌组织中表达上调，6例（10.3%）表达下调。结直肠癌组织中hsa_circ_0078607的相对表达量与肿瘤位置（P=0.029）、分化程度（P=0.046）和远处转移（P=0.043）有关联。ROC结果显示，在结直肠癌组织和血清中其诊断结直肠癌的AUC分别为0.845 7[95%CI（0.772 8，0.918 6），P<0.000 1]和0.868 3[95%CI（0.790 7，0.945 9），P<0.000 1]；在息肉患者血清中，hsa_circ_0078607诊断结直肠息肉的AUC为0.710 1 [95%CI（0.610 0，0.810 1）]。GO/KEGG富集分析结果表明，hsa_circ_0078607下游的miRNA可能参与RNA聚合酶Ⅱ启动子转录调控、蛋白K48-连锁泛素化、Wnt、Hippo及MAPK信号通路调控等多个生物过程。结论：Hsa_circ_0078607在结直肠癌细胞、组织和血清中呈高表达，其在结直肠癌组织中的表达水平与肿瘤位置、分化程度和远处转移有关联，提示其可作为结直肠癌潜在的分子诊断标志物；其还可能介导结直肠癌的发生发展过程，对发现结直肠癌潜在的治疗靶点有重要意义。

The Drosha-Independent MicroRNA6778-5p/GSK3ß Axis Mediates the Proliferation of Gastric Cancer Cells.

Background: Gastric cancer (GC) is a primary cause of cancer death around the world. Previous studies have found that Drosha plays a significant role in the development of tumor cells. Soon after, we unexpectedly found that the expression of microRNA6778-5p (miR6778-5p) is unconventionally high in the gastric cancer cells low-expressing Drosha. So, we designed the Drosha interference sequence and recombined it into a lentiviral vector to construct Drosha knockdown lentivirus and transfected the Drosha knockdown lentivirus into gastric cancer cells to establish Drosha knockdown gastric cancer cell lines. We aimed to explore the effect of microRNA6778-5p on the proliferation of gastric cancer cells with Drosha knockdown and its intrinsic mechanism. Methods: We designed the Drosha interference sequence and recombined it into a lentiviral vector to construct Drosha knockdown lentivirus and transfected the Drosha knockdown lentivirus into gastric cancer cells to establish Drosha knockdown gastric cancer cell lines. After transfecting miR6778-5p mimics and inhibitor into gastric cancer cell lines with Drosha knockdown, the expression levels of miR6778-5p mimics in Drosha low-expressing gastric cancer cells increased, while miR6778-5p inhibitor decreased the expression levels of miR6778-5p. The Cell Counting Kit-8 (CCK-8) experiment was used to detect the proliferation ability of gastric cancer cells after overexpression or knockdown of miR6778-5p and bioinformatics predicted the relationship between miR6778-5p and glycogen synthase kinase-3ß (GSK3ß). Results: After infection with the Drosha knockdown lentivirus, Drosha's mRNA and protein levels were significantly downregulated in gastric cancer cells. The expression levels of miR6778-5p mimics in Drosha low-expressing gastric cancer cells increased, while miR6778-5p inhibitor decreased the expression levels of miR6778-5p. Overexpression of miR6778-5p significantly enhanced the proliferation ability of Drosha low-expression gastric cancer cells; on the contrary, knocking down miR6778-5p weakened the proliferation ability of Drosha low-expression gastric cancer cells. Bioinformatics predicted that miR6778-5p targeted glycogen synthase kinase-3ß (GSK3ß) and the mRNA and protein levels of GSK3ß decreased significantly after overexpression of miR6778-5p. Conclusion: miR6778-5p promotes the proliferation of Drosha low-expressing gastric cancer cells by targeting GSK3ß.

Diagnostic Value of Serum DNASE1L3 in Hepatitis B Virus-Related Hepatocellular Carcinoma.

BACKGROUND: Deoxyribonuclease 1-like 3 (DNASE1L3) is an endonuclease associated with many autoimmune diseases and tumors. However, the serum DNASE1L3 level in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) remains unreported. Thus, this study compared the diagnostic value of DNASE1L3 and alpha-feto-protein (AFP) individually and in combination in HBV-related HCC. METHODS: The study population consisted of 88 patients with HBV-related HCC, 80 patients with HBV-related liver cirrhosis (LC) and 88 control subjects. The serum DNASE1L3 levels were measured using an enzyme-linked immunosorbent assay. The serum AFP was also assayed. RESULTS: Our data showed that the serum DNASE1L3 levels were significantly higher in patients with HBV-related HCC than in the healthy controls and patients with LC. When the two biomarkers were analyzed individually, the receiver operating characteristic curve analysis showed that the areas under the curve of DNASE1L3 and AFP were 0.898 and 0.866, respectively. When DNASE1L3 and AFP were combined, the area under the curve was 0.951. The sensitivities of DNASE1L3 and AFP were 72.73% and 74.81%, respectively, and the specificities were 93.18% and 92.05%, respectively, in the diagnosis of HBV-related HCC. The sensitivity of the two combined could be improved to 89.77%. However, no correlation was found between serum DNASE1L3 and AFP in HBV-related HCC patients (r = 0.005, p = 0.734). CONCLUSIONS: Serum DNASE1L3 has high sensitivity and specificity in the diagnosis of HCC. DNASE1L3 combined with AFP has higher sensitivity and can improve the diagnostic efficiency of HBV-related HCC.

Clinical application of red cell distribution width, mean platelet volume, and cancer antigen 125 detection in endometrial cancer.

BACKGROUND: Red cell distribution width (RDW) and mean platelet volume (MPV) are considered to be associated with tumors. We investigated the diagnostic value of RDW, MPV, and cancer antigen 125 (CA125), alone or in combination, in the diagnosis of endometrial cancer and endometrial hyperplasia. METHODS: This study included 144 patients with endometrial cancer (stage I: 32; II: 42; III: 48; and IV: 22), 104 patients with endometrial hyperplasia, and 80 healthy control subjects. The whole blood cell parameters were analyzed by a Mindray Blood Cell Analyzer (CAL8000), whereas CA125 was analyzed using an Architect i2000 Analyzer (Abbott). RESULTS: Significant differences in RDW, MPV, and CA125 level were observed in the endometrial cancer, endometrial hyperplasia, and control groups (P < .05). Red cell distribution width was positively correlated (r = .735) whereas MPV was negatively correlated with (r = -.736) endometrial cancer staging. The area under the receiver operating characteristic curve of the combined diagnosis of endometrial cancer based on RDW, MPV, and CA125 was 0.924 (95% CI: 0.881-0.955). The sensitivity and specificity of the combined diagnosis were larger than those of the independent detections involving RDW, MPV, and CA125. CONCLUSIONS: The combination of RDW, MPV, and CA125 can improve the differential diagnosis of endometrial cancer and endometrial hyperplasia.

Oxidized ATM-mediated glycolysis enhancement in breast cancer-associated fibroblasts contributes to tumor invasion through lactate as metabolic coupling.

BACKGROUND: Cancer-associated fibroblasts (CAFs) are the predominant residents in the breast tumor microenvironment. In our work, we found activation of DNA damage-independent ATM (oxidized ATM), enhanced glycolysis and aberrant metabolism-associated gene expressions in breast CAFs. Nevertheless, whether and how oxidized ATM regulates the glycolytic activity of CAFs keep in unveil. Recently, a reverse Warburg effect was observed in tumor tissues, in which host cells (such as CAFs, PSCs) in the tumor microenvironment have been found to "fuel" the cancer cells via metabolites transfer. However, the molecular mechanisms of the metabolites from stromal cells playing a role to the progression of cancer cells remain to be determined. METHODS: Oxidized ATM activation in stromal CAFs was assessed by western blotting and immunofluorescence. The increased glycolytic ability of CAFs was validated by measurements of OCR and ECAR and detections of glucose consumption and lactate production. Kinase assay and western blotting were performed to confirm the phosphorylation of GLUT1. The membrane location of phosphorylated GLUT1 was determined by biotin pull-down assay and immunofluorescence staining. The regulation of PKM2 through oxidized ATM was evaluated by western blots. In addition, the impact of lactate derived from hypoxic CAFs on cancer cell invasion was investigated both in vitro (transwell assays, western blots) and in vivo (orthotopic xenografts). FINDINGS: Hypoxia-induced oxidized ATM promotes glycolytic activity of CAFs by phosphorylating GLUT1 at S490 and increasing PKM2 expression. Moreover, lactate derived from hypoxic CAFs, acting as a metabolic coupling between CAFs and breast cancer cells, promotes breast cancer cell invasion by activating the TGFß1/p38 MAPK/MMP2/9 signaling axis and fueling the mitochondrial activity in cancer cells. INTERPRETATION: Our work shows that oxidized ATM-mediated glycolysis enhancement in hypoxic stromal fibroblasts plays an essential role in cancer cell invasion and metastasis and may implicate oxidized ATM as a target for breast tumor treatment. FUND: This research was supported by National Natural Science Foundation of China.

Application of molecular, microbiological, and immunological tests for the diagnosis of bone and joint tuberculosis.

BACKGROUND: To evaluate the application of interferon gamma release assay (IGRA), rifampicin resistant real-time fluorescence quantitative PCR technique Xpert Mycobacterium tuberculosis/rifampicin (Xpert MTB/RIF), and the levels of TNF-α and TGF-ß in the diagnosis of bone and joint tuberculosis. METHODS: Eighty-six patients with bone and joint tuberculosis, diagnosed by pathology or microbiology, were examined by Xpert MTB/RIF and IGRA (T-SPOT. TB) for Mycobacterium tuberculosis infection, and the TNF-α and TGF-ß levels of the patients were measured. RESULTS: The sensitivity of IGRA in diagnosing bone and joint tuberculosis was 81.4%; Xpert MTB/RIF's sensitivity was 70.9%. The combined sensitivity of the two methods was 91.9%. The combined detection sensitivity of the two methods was higher than individual IGRA or Xpert MTB/RIF detection sensitivity. The TNF-α and TGF-ß levels in bone and joint tuberculosis patients were higher than those in the control group. CONCLUSION: Xpert MTB/RIF, IGRA, TNF-α, and TGF-ßs expression have value in the rapid diagnosis of bone and joint tuberculosis, and the sensitivity and accuracy of bone and joint tuberculosis diagnosis by combining them can improve it.

Elevated serum visfatin levels are associated with poor prognosis of hepatocellular carcinoma.

Visfatin is considered a pro-inflammatory adipocytokine, and it is commonly increased in obesity-related diseases. This study aimed to evaluate the levels of serum visfatin in patients with hepatocellular carcinoma (HCC) and its diagnostic and predictive value in detecting HCC. Fasting serum levels of visfatin of 135 HCC patients, 115 chronic hepatitis B (CHB) patients, 129 liver cirrhosis (LC) patients, and 149 healthy controls were determined via enzyme-linked immunosorbent assay. Meanwhile, serum alpha fetal protein (AFP) and interleukin-6 (IL-6) were also assayed. The median serum visfatin concentration in HCC patients was 1.113 ng/mL (range: 0.823-2.214 ng/mL), which was significant higher than those of healthy controls, CHB patients, and LC patients (P<0.05). The serum visfatin concentration in HCC patients was positively correlated with AFP (r=0.595, P<0.001) and IL-6 (r=0.261, P<0.015) and was also associated with tumor size and tumor node metastasis stage. Moreover, elevated levels of serum visfatin were associated with a higher HCC risk for CHB and LC patients. Multivariate Cox regression analysis had shown that HCC patients with high levels of serum visfatin had significantly shorter overall survival times than those with low serum visfatin levels (P<0.001). Using a cutoff visfatin level of 1.403 ng/mL, the receiver operating characteristic curve analysis showed unappealing sensitivity and specificity values (45.76% and 74.79%, respectively; AUC=0.626) regarding visfatin's use as a diagnostic marker for HCC. Our results indicate that increased serum visfatin levels are associated with poor prognosis of HCC. Visfatin may be a potential therapeutic target of HCC.

Genetic variant in visfatin gene promoter contributes to reduced risk of hepatocellular carcinoma in a Chinese population.

Knowledge on the role of gene variants in the visfatin promoter region in the hepatitis B virus (HBV)-related liver diseases is limited. In this study, we genotyped two potentially functional single nucleotide polymorphisms (SNPs) in the visfatin promoter region, -1535C>T (rs61330082) and -3187G>A (rs11977021), in 120 HBV-related chronic hepatitis B (CHB) patients, 140 HBV-related liver cirrhosis (HBV-LC) patients, 243 HBV-related hepatocellular carcinoma (HBV-HCC) patients, and 224 asymptomatic HBV carriers. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated by logistic regression. The results showed subjects with a TT genotype of -1535C>T had a significantly decreased risk of HBV-HCC related to the CC and CC + CT genotypes (adjusted OR = 0.493, 95% CI = 0.313-0.778; OR = 0.535, 95% CI = 0.362-0.791, respectively). A lowered risk also appeared in the comparison between allele T and allele C (OR = 0.734, 95%, CI = 0.581-0.950). However, these associations existed only in people with Zhuang ethnicity, but not in people with Han ethnicity. There were no significant associations between -3187G>A polymorphisms and the risk of HBV-related liver diseases. Our results suggested that visfatin -1535C>T polymorphisms might be associated with decreased risk of HBV-HCC among the ethnic Zhuang population in Guangxi, China.

CA153 in Breast Secretions as a Potential Molecular Marker for Diagnosing Breast Cancer: A Meta Analysis.

PURPOSE: Many studies have reported that carbohydrate antigen 153 (CA153) in breast secretions (BS) can discriminate breast cancer (BC) patients from healthy individuals, indicating CA153 in BS as a potential index for BC. This meta-analysis aimed to evaluate the actual diagnostic value of CA153 in BS. METHODS: Related papers were obtained from Pubmed, Embase, Scopus, Ovid, Sciverse, the Cochrane library, Chinese Biomedical literature Database (CBM), Technology of Chongqing (VIP), Wan Fang Data, and Chinese National Knowledge Infrastructure (CNKI). Pooled sensitivity, specificity, and diagnostic odds ratio (DOR) of CA153 in BS for BC diagnosis were analyzed with the random effect model. SROC and the area under the curve (AUC) were applied to assess overall diagnostic efficiency. RESULTS: This meta-analysis included five studies with a total of 329 BC patients and 381 healthy subjects. For CA153 in BS, the summary sensitivity, specificity, and DOR to diagnose BC were 0.63 (95% confidence interval (CI): 0.57∼0.68), 0.82 (95% CI: 0.78∼0.86), and 9.18 (95% CI: 4.22∼19.95), respectively. Furthermore, the AUC of BS CA153 in the diagnosis of BC was 0.8614. CONCLUSIONS: CA153 in BS is a valuable molecular marker in diagnosing BC and should be applied in standard clinical practices of BC screening upon confirmation of our findings in a larger prospective study.

Association of Interferon Gamma +874T/A Polymorphism and Leukemia Risk: A Meta-Analysis.

Interferon gamma (IFN-γ) has antitumor and antiproliferative effects, and previous studies indicated IFN-γ +874T/A (rs2430561) polymorphism were related to the risk of many types of cancer. However, the association between IFN-γ +874T/A polymorphism and leukemia risk remained controversial.We performed a comprehensive meta-analysis based on the Preferred Reporting Items for Systematic Reviews and Meta-analyses statement (PRISMA). Electronic database of Embase, Pubmed, and the Cochrane Library were searched for eligible articles published up to December 13, 2015. The association between genetic polymorphisms and leukemia risk was measured by odds ratios (ORs) and its corresponding 95% confidence intervals (CIs).A total of 8 studies amounting to 420 patients and 767 control subjects were retrieved for this study. Although associations between IFN-γ +874T/A polymorphism and overall leukemia risks were lacking, decreased chronic lymphocytic leukemia (CLL) risk was detected in the allelic model (T vs A, OR=0.660, 95%CI = 0.483-0.902, P = 0.009, I = 0.0% and P = 0.863 for heterogeneity), the codominant model (TT vs AA, OR = 0.472, 95%CI = 0.247-0.902, P = 0.023, I = 0.0% and P = 0.994 for heterogeneity), and dominant model (TT + TA vs AA, OR = 0.457, 95%CI = 0.285-0.734, P = 0.001, I = 40.3% and P = 0.195 for heterogeneity) by using fixed-effect model separately. On the contrary, results indicated T carries have an increased chronic myelogenous leukemia (CML) risk in dominant model (TT + TA vs AA, OR = 1.783, 95%CI = 1.236-2.573, P = 0.002, I = 19.0% and P = 0.295 for heterogeneity).This study suggests IFN-γ +874T/A polymorphism are related to CML and CLL risk. In addition, our work also points out IFN-γ +874T/A polymorphism may play dual contrasting role in leukemia risk.

CEA in breast ductal secretions as a promising biomarker for the diagnosis of breast cancer: a systematic review and meta-analysis.

PURPOSE: Other studies have shown that levels of carcinoembryonic antigen (CEA) in breast ductal secretions (BDS) differ significantly between breast cancer (BC) patients and healthy individuals, providing direct evidence for CEA in BDS as a promising biomarker for BC. This meta-analysis was designed to assess the potential diagnostic value of CEA in BDS. METHODS: Relevant articles were retrieved from Embase, Pubmed, and the Cochrane Library. Sensitivity, specificity, and diagnostic odds ratio (DOR) of CEA in BDS for diagnosing BC were pooled using random effects models. SROC and the area under the curve (AUC) were used to estimate overall diagnostic performance. RESULTS: This meta-analysis comprised five studies with a total of 340 BC patients and 448 healthy controls. For CEA in BDS, the pooled sensitivity, specificity, and DOR to diagnose BC were 58 % [95 % confidence interval (CI): 52-63 %], 87 % (95 % CI: 84-90 %), and 7.07 (95 % CI: 3.10-16.12), respectively. Moreover, the AUC of CEA in the diagnosis of BC was 0.8570. CONCLUSIONS: CEA in BDS is a promising biomarker in the diagnosis of BC and should be evaluated as a standard screening tool upon verification of our results in a larger study population.

Comparison of Architect i2000sr and Cobas e601 Systems for Determining Serum Human Chorionic Gonadotropin-Beta.

BACKGROUND: Human chorionic gonadotropin-beta (ß-hCG) is an important index used to monitor embryonic development following embryo transfer. Architect i2000sr and Cobas e601 are widely used automated immunoassay systems used to measure serum ß-hCG concentrations; however, the correlations between serum ß-hCG levels measured with these two immunoassays and the accuracy of the immunoassays have not been fully evaluated. METHODS: Serum ß-hCG levels were measured in 133 serum samples using the Architect i2000sr and Cobas e601 automated immunoassay systems. Passing-Bablok regression analysis was used to compare the correlation in serum ß-hCG levels obtained using the two immunoassays. A Bland-Altman plot analysis was used to identify mean ratios and 95% CIs of the mean ratios of the ß-hCG results between the two immunoassays. In this graphical method the mean ratios between the two techniques were plotted against the averages of the two techniques. RESULTS: The total coefficients of variations (CVs) of serum ß-hCG ranged from 3.12 - 4.66% for Cobas e601 and 3.18 - 4.99% for Architect i2000sr. The measured value of serum ß-hCG detected by the two immunoassays was statistically significant (p < 0.001). The Passing-Bablok regression analysis showed good correlation between the serum ß-hCG values measured using the two systems. At a low concentration of serum ß-hCG (< 10000 IU/L, n = 52), the correlation coefficient r was 0.9628. At a high concentration of serum ß-hCG (> 10000 IU/L, n = 81), the correlation coefficient r was 0.8076. The Bland-Altman plot analysis showed that the measured value of serum ß-hCG detected by Architect i2000sr was about 1.25 times higher than that of Cobas e601. The mean ratio was 1.12 at a low concentration of serum ß-hCG, and it was 1.33 at a high concentration. CONCLUSIONS: Architect i2000sr and Cobas e601 have good concordance for determining serum ß-hCG. However, the ß-hCG values measured with Architect i2000sr were 25% higher than those obtained using Cobas e601.

LncRNA-Hh Strengthen Cancer Stem Cells Generation in Twist-Positive Breast Cancer via Activation of Hedgehog Signaling Pathway.

Cancer stem cells (CSCs) are a subpopulation of neoplastic cells with self-renewal capacity and limitless proliferative potential as well as high invasion and migration capacity. These cells are commonly associated with epithelial-mesenchymal transition (EMT), which is also critical for tumor metastasis. Recent studies illustrate a direct link between EMT and stemness of cancer cells. Long non-coding RNAs (lncRNAs) have emerged as important new players in the regulation of multiple cellular processes in various diseases. To date, the role of lncRNAs in EMT-associated CSC stemness acquisition and maintenance remains unclear. In this study, we discovered that a set of lncRNAs were dysregulated in Twist-positive mammosphere cells using lncRNA microarray analysis. Multiple lncRNAs-associated canonical signaling pathways were identified via bioinformatics analysis. Especially, the Shh-GLI1 pathway associated lncRNA-Hh, transcriptionally regulated by Twist, directly targets GAS1 to stimulate the activation of hedgehog signaling (Hh). The activated Hh increases GLI1 expression, and enhances the expression of SOX2 and OCT4 to play a regulatory role in CSC maintenance. Thus, the mammosphere-formation efficiency (MFE) and the self-renewal capacity in vitro, and oncogenicity in vivo in Twist-positive breast cancer cells are elevated. lncRNA-Hh silence in Twist-positive breast cells attenuates the activated Shh-GLI1 signaling and decreases the CSC-associated SOX and OCT4 levels, thus reduces the MFE and tumorigenesis of transplanted tumor. Our results reveal that lncRNAs function as an important regulator endowing Twist-induced EMT cells to gain the CSC-like stemness properties.

High-visfatin levels in women with polycystic ovary syndrome: evidence from a meta-analysis.

OBJECTIVE: Previous studies regarding visfatin levels in women with polycystic ovary syndrome (PCOS) showed conflicting results. To evaluate the visfatin levels in PCOS, a meta-analysis was performed. METHODS: A comprehensive literature search of eligible studies in Embase, Pubmed and the Cochrane Library was undertaken through November 2014. Standardized mean differences (SMDs) with 95% confidence intervals (CIs) were calculated to estimate the strength of the association. RESULTS: A total of 1341 subjects (695 cases and 646 controls) were included in this meta-analysis. The pooled analysis results indicated that the visfatin levels were significantly higher in PCOS patients than that of controls (SMD = 1.19, 95% CI 0.77-1.60, p = 0.000). The results from stratified analysis and univariate analysis suggested that high-visfatin levels were not related to body mass index (BMI), insulin resistance (IR) and total testosterone ratio. Significant heterogeneity was observed in all analysis. CONCLUSION: Our results indicate that high-circulating visfatin level is an intrinsic characteristic of PCOS, which suggests visfatin could be a potential biomarker for PCOS.

Twist promotes reprogramming of glucose metabolism in breast cancer cells through PI3K/AKT and p53 signaling pathways.

Twist, a key regulator of epithelial-mesenchymal transition (EMT), plays an important role in the development of a tumorigenic phenotype. Energy metabolism reprogramming (EMR), a newly discovered hallmark of cancer cells, potentiates cancer cell proliferation, survival, and invasion. Currently little is known about the effects of Twist on tumor EMR. In this study, we found that glucose consumption and lactate production were increased and mitochondrial mass was decreased in Twist-overexpressing MCF10A mammary epithelial cells compared with vector-expressing MCF10A cells. Moreover, these Twist-induced phenotypic changes were augmented by hypoxia. The expression of some glucose metabolism-related genes such as PKM2, LDHA, and G6PD was also found to be upregulated. Mechanistically, activated ß1-integrin/FAK/PI3K/AKT/mTOR and suppressed P53 signaling were responsible for the observed EMR. Knockdown of Twist reversed the effects of Twist on EMR in Twist-overexpressing MCF10A cells and Twist-positive breast cancer cells. Furthermore, blockage of the ß1-integrin/FAK/PI3K/AKT/mTOR pathway by siRNA or specific chemical inhibitors, or rescue of p53 activation can partially reverse the switch of glucose metabolism and inhibit the migration of Twist-overexpressing MCF10A cells and Twist-positive breast cancer cells. Thus, our data suggest that Twist promotes reprogramming of glucose metabolism in MCF10A-Twist cells and Twist-positive breast cancer cells via activation of the ß1-integrin/FAK/PI3K/AKT/mTOR pathway and inhibition of the p53 pathway. Our study provides new insight into EMR.

AXIN2 rs2240308 polymorphism contributes to increased cancer risk: evidence based on a meta-analysis.

BACKGROUND: Variants in the axis inhibition 2 (AXIN2) gene might alter the protein's structure or function or create a multiprotein destruction complex in the Wnt signaling pathway and thus affect an individual's susceptibility to cancer. The objective of this study is to evaluate broadly the evidence available for the AXIN2 rs2240308 polymorphism and risk of cancer. METHODS: A comprehensive literature search was undertaken for eligible studies in Embase, PubMed, and Cochrane Library up to Nov 30, 2014. Odds ratios (ORs) and the corresponding 95 % confidence intervals (CIs) were used to measure the strength of the models. RESULTS: Eight articles (10 case-control studies with 1,502 cases and 1,590 controls) were included in this analysis. Overall, the AXIN2 rs2240308 polymorphism was associated with a significant increase in the risk of cancer (G allele vs. A allele: OR = 1.21, 95 % CI = 1.05-1.40, I (2) = 39.5 % and P Q = 0.094 for heterogeneity; GG vs. AA: OR = 1.30, 95 % CI = 1.04-1.63, I (2) = 35.9 % and P Q = 0.121 for heterogeneity; GG vs. GA + AA: OR = 1.36, 95 % CI = 1.17-1.58, I (2) = 19.5 % and P Q = 0.263 for heterogeneity). Asian populations showed similar results. Stratified analysis by cancer types indicated that the AXIN2 rs2240308 polymorphism increases the risk of lung cancer (G allele vs. A allele: OR = 1.36, 95 % CI = 1.17-1.59; GA vs. AA: OR = 1.43, 95 % CI = 1.01-2.02; GG vs. AA: OR = 1.93, 95 % CI = 1.36-2.75; GG + GA vs. AA: OR = 1.65, 95 % CI = 1.18-2.30; GG vs. GA + AA: OR = 1.45, 95 % CI = 1.18-1.79. All I (2) < 50 % and P Q > 0.100 for heterogeneity). CONCLUSIONS: This study showed that the AXIN2 rs2240308 polymorphism contribute to increasing the risk of cancer, especially lung cancer in Asian populations.

Oxidized ATM promotes abnormal proliferation of breast CAFs through maintaining intracellular redox homeostasis and activating the PI3K-AKT, MEK-ERK, and Wnt-ß-catenin signaling pathways.

Abnormal proliferation is one characteristic of cancer-associated fibroblasts (CAFs), which play a key role in tumorigenesis and tumor progression. Oxidative stress (OS) is the root cause of CAFs abnormal proliferation. ATM (ataxia-telangiectasia mutated protein kinase), an important redox sensor, is involved in DNA damage response and cellular homeostasis. Whether and how oxidized ATM regulating CAFs proliferation remains unclear. In this study, we show that there is a high level of oxidized ATM in breast CAFs in the absence of double-strand breaks (DSBs) and that oxidized ATM plays a critical role in CAFs proliferation. The effect of oxidized ATM on CAFs proliferation is mediated by its regulation of cellular redox balance and the activity of the ERK, PI3K-AKT, and Wnt signaling pathways. Treating cells with antioxidant N-acetyl-cysteine (NAC) partially rescues the proliferation defect of the breast CAFs caused by ATM deficiency. Administrating cells with individual or a combination of specific inhibitors of the ERK, PI3K-AKT, and Wnt signaling pathways mimics the effect of ATM deficiency on breast CAF proliferation. This is mainly ascribed to the ß-catenin suppression and down-regulation of c-Myc, thus further leading to the decreased cyclinD1, cyclinE, and E2F1 expression and the enhanced p21(Cip1) level. Our results reveal an important role of oxidized ATM in the regulation of the abnormal proliferation of breast CAFs. Oxidized ATM could serve as a potential target for treating breast cancer.