Progesterone receptor (PR) expression correlates strongly with progestin sensitivity in fertility-sparing therapy for endometrial carcinoma (EC). However, the mechanisms governing PR expression remain incompletely defined. Here, by stratifying EC patients into PR-high and PR-low groups, we observe that PR-low tumors exhibit enhanced invasion and metastasis signatures, whereas PR-high tumors display increased fatty acid metabolism. Through integrated network analysis, transcriptional correlation across multiple cohorts, and single-cell transcriptomic profiling, FOXA2 is identified as a master regulator of PR expression. Specifically, FOXA2 directly binds the PR promoter, which, in turn, transcriptionally activates PR expression and increases the sensitivity of EC cells to medroxyprogesterone acetate (MPA), an oral progestin used in clinical. Overexpression of FOXA2 markedly inhibits tumor progression, evidenced with reduced cell proliferation and migration while elevated apoptosis. Moreover, FOXA2 is critically involved in lipid metabolic modulation and the administration of Orlistat, an FDA-approval inhibitor of fatty acid synthase, elevates FOXA2 and PR expression, subsequently enhancing progestin sensitivity both in vitro and in vivo. Collectively, our findings identify FOXA2 as a key regulator in controlling PR levels in EC cells and propose the activation of FOXA2-PR axis via Orlistat treatment as a promising therapeutic strategy to improve progestin responsiveness in EC patients.

Background: Endometrial carcinoma is one of the most common female malignancies worldwide. Based on our preliminary investigation, DUSP1 was identified as a potential biomarker for endometrial carcinoma prognosis, but its function and mechanism remained unclear. Methods: In this study, genes highly correlated with DUSP1 in endometrial cancer were found through correlation analysis, and the promoter sequence of DUSP1 was analyzed by PROMO program. Next-generation phosphorylation mass spectrometry was used to explore new downstream target proteins and pathways of DUSP1 in endometrial carcinoma. The mRNA and protein expression levels were detected by real-time quantitative PCR, immunohistochemistry and Western blotting. The cell survival and proliferation were analyzed by CCK8 assay, cell apoptosis was analyzed by Annexin-V-APC and PI dual staining assay, and the cell invasion was analyzed by Transwell method. Results: (1) There was a high correlation between the expression of DUSP1 and the genes involved in AP-1 complex and its co-expression network. (2) Promoter sequence analysis predicted that the members of AP-1 complex might be the upstream transcriptional regulators of DUSP1. (3) Transfection experiments proved DUSP1 can inhibit tumor growth and invasion, and promote apoptosis by regulating ERK pathway. (4) The results of phosphorylation mass spectrometry showed that overexpression of DUSP1 mainly dephosphorylated EPHA2 in endometrial carcinoma, and co-immunoprecipitation verified the protein interaction between DUSP1 and EPHA2. (5) Overexpression or knockdown of EPHA2 significantly changed the phosphorylation level of EPHA2. (6) The expression of EPHA2 protein was high in patients with more aggressive endometrial cancer. (7) Using EPHA2 inhibitor could significantly slow down the growth rate of tumor cells. Conclusion: (1) There exists a mutual regulation relationship between DUSP1 and AP-1 co-expression network in endometrial carcinoma. (2) It is reported for the first time that DUSP1 phosphatase acts on the ser899 site of EphA2 in endometrial carcinoma. (3) DUSP1 can inhibit tumor growth and invasion, and promote apoptosis by regulating MAPK pathway through directly dephosphorylating ERK, or by dephosphorylating EPHA2.

Background: Steroid receptor-associated and regulated protein (SRARP) suppresses tumor progression and modulates steroid receptor signaling by interacting with estrogen receptors and androgen receptors in breast cancer. In endometrial cancer (EC), progesterone receptor (PR) signaling is crucial for responsiveness to progestin therapy. The aim of this study was to investigate the role of SRARP in tumor progression and PR signaling in EC.Methods: Ribonucleic acid sequencing data from the Cancer Genome Atlas, Clinical Proteomic Tumor Analysis Consortium, and Gene Expression Omnibus were used to analyze the clinical significance of SRARP and its correlation with PR expression in EC. The correlation between SRARP and PR expression was validated in EC samples obtained from Peking University People's Hospital. SRARP function was investigated by lentivirus-mediated overexpression in Ishikawa and HEC-50B cells. Cell Counting Kit-8 assays, cell cycle analyses, wound healing assays, and Transwell assays were used to evaluate cell proliferation, migration, and invasion. Western blotting and quantitative real-time polymerase chain reaction were used to evaluate gene expression. The effects of SRARP on the regulation of PR signaling were determined by co-immunoprecipitation, PR response element (PRE) luciferase reporter assay, and PR downstream gene detection.Results: Higher SRARP expression was significantly associated with better overall survival and disease-free survival and less aggressive EC types. SRARP overexpression suppressed growth, migration, and invasion in EC cells, increased E-cadherin expression, and decreased N-cadherin and Wnt family member 7A ( WNT7A ) expression. SRARP expression was positively correlated with PR expression in EC tissues. In SRARP -overexpressing cells, PR isoform B (PRB) was upregulated and SRARP bound to PRB. Significant increases in PRE-based luciferase activity and expression levels of PR target genes were observed in response to medroxyprogesterone acetate.Conclusions: This study illustrates that SRARP exerts a tumor-suppressive effect by inhibiting the epithelial-mesenchymal transition via Wnt signaling in EC. In addition, SRARP positively modulates PR expression and interacts with PR to regulate PR downstream target genes.

Transient receptor potential vanilloid 4 (TRPV4) is a calcium-permeable cation channel that has been associated with several types of cancer. However, its biological significance, as well as its related mechanism in endometrial cancer (EC) still remains elusive. In this study, we examined the function of calcium in EC, with a specific focus on TRPV4 and its downstream pathway. We reported here on the findings that a high level of serum ionized calcium was significantly correlated with advanced EC progression, and among all the calcium channels, TRPV4 played an essential role, with high levels of TRPV4 expression associated with cancer progression both in vitro and in vivo. Proteomic and bioinformatics analysis revealed that TRPV4 was involved in cytoskeleton regulation and Rho protein pathway, which regulated EC cell migration. Mechanistic investigation demonstrated that TRPV4 and calcium influx acted on the cytoskeleton via the RhoA/ROCK1 pathway, ending with LIMK/cofilin activation, which had an impact on F-actin and paxillin (PXN) levels. Overall, our findings indicated that ionized serum calcium level was significantly associated with poor outcomes and calcium channel TRPV4 should be targeted to improve therapeutic and preventive strategies in EC.

Cell adhesion molecules (CAMs) determine the behavior of cancer cells during metastasis. Although some CAMs are dysregulated in certain types of cancer and are associated with cancer progression, to the best of our knowledge, a comprehensive study of CAMs has not been undertaken, particularly in endometrial cancer (EC). In the present study the expression of 225 CAMs in EC patients with various clinicopathological phenotypes were evaluated by statistical analysis using publicly available data from The Cancer Genome Atlas database. The Kaplan-Meier method, and univariate and multivariate Cox proportional hazards regression models were used for survival analyses. Among the differentially expressed CAMs that were associated with aggressive clinicopathological phenotypes, 10 CAM genes were independent prognostic factors compared with other clinicopathological prognostic factors, including stage, grade, age, lymph node status, peritoneal cytology and histological subtype. A total of six genes (L1 cell adhesion molecule, mucin 15, cell surface associated, cell adhesion associated, oncogene regulated, immunoglobulin superfamily member 9B, protocadherin 9 and protocadherin β1) were selected for integrative analysis. The six-gene signature was demonstrated to be an independent prognostic factor and could effectively stratify patients with different risks. Patients with more high-expression CAMs had a higher risk of poor overall survival (OS) rate. The mortality risk for patients with elevation of >4 CAMs was 11 times of that in those without elevation of these 6 CAMs. Similar results were obtained when relapse-free survival (RFS) time was used during the analysis. Prognostic reliability of the six-gene model was validated using data of an independent cohort from the International Cancer Genome Consortium. In conclusion, a combination of CAM alterations contributed to progression and aggressiveness of EC. The six-gene signature was effective for predicting worse OS and RFS in patients with EC and could be complementary to the present clinical prognostic criteria.

Traditional clinical features are not sufficient to accurately judge the prognosis of endometrioid endometrial adenocarcinoma (EEA). Molecular biological characteristics and traditional clinical features are particularly important in the prognosis of EEA. The aim of the present study was to establish a predictive model that considers genes and clinical features for the prognosis of EEA. The clinical and RNA sequencing expression data of EEA were derived from samples from The Cancer Genome Atlas (TCGA) and Peking University People's Hospital (PKUPH; Beijing, China). Samples from TCGA were used as the training set, and samples from the PKUPH were used as the testing set. Variable selection using Random Forests (VSURF) was used to select the genes and clinical features on the basis of TCGA samples. The RF classification method was used to establish the prediction model. Kaplan-Meier curves were tested with the log-rank test. The results from this study demonstrated that on the basis of TCGA samples, 11 genes and the grade were selected as the input features. In the training set, the out-of-bag (OOB) error of RF model-1, which was established using the ‘11 genes’, was 0.15; the OOB error of RF model-2, which was established using the ‘grade’, was 0.39; and the OOB error of RF model-3, established using the ‘11 genes and grade’, was 0.15. In the testing set, the classification accuracy of RF model-1, model-2 and model-3 was 71.43, 66.67 and 80.95%, respectively. In conclusion, to the best of our knowledge, the VSURF was used to select features relevant to EEA prognosis, and an EEA predictive model combining genes and traditional features was established for the first time in the present study. The prediction accuracy of the RF model on the basis of the 11 genes and grade was markedly higher than that of the RF models established by either the 11 genes or grade alone.

Kallikrein-related peptidase (KLK) family is one of the major serine proteases in tumor microenvironment, which plays a crucial role in cancer invasion and metastasis. A number of KLK family members have been found to be upregulated or downregulated in some cancers, and some KLKs may be potential biomarkers for cancers. However, little is known about the role of KLKs in endometrial carcinoma (EC). In this study, we analyzed the mRNA sequencing data of EC from The Cancer Genome Atlas (TCGA) public database and found that the higher expression of KLK family members 5-8 (KLK5-8) was associated with an aggressive clinicopathologic phenotype and worse prognosis in EC patients. High expression of KLK5-8 was also confirmed in our patients with advanced stage and high-grade EC, as well as in a highly invasive cell line. Our study also demonstrated the differences between the subcellular localization of KLK5-8 and the co-expression of different splicing variants of KLK5-8 in EC cells, suggesting that various isoforms of KLK5-8 may work synergistically to regulate invasion and migration. We found that the elevation of more KLKs in a patient's sample indicated a higher risk of worse survival. Combination of KLK5-8 was shown to be an independent prognostic factor for overall survival by multivariate Cox regression (hazard ratio: 2.215, 95% confidence interval: 1.045-4.694, P=0.038), and may be a promising biomarker.

纺锤体关卡是一个在进化上高度保守的有丝分裂监督系统,确保染色体在有丝分裂过程中高度保真.丝氨酸/苏氨酸蛋白激酶BUB1作为BUB家族成员之一,是纺锤体关卡的重要组成成分.BUB1表达异常,会使纺锤体关卡功能缺陷,导致染色体不稳定性及非整倍体细胞的发生.

BACKGROUND:Management of tumors has become more complex owing to tumor heterogeneity. Fewer studies have been performed on intra-tumor heterogeneity of endometrial cancer (EC) until now. Therefore, it is of great clinical value to explore the intra-tumor heterogeneity of EC based on clinical features and gene expression profiles. METHODS:A total of 1688 patients with EC were screened and 114 patients were finally selected, including specimens from 84 patients with primary EC without relapse (PE) and the paired metastases (P-M) specimens, as well as specimens from 30 patients with primary EC with relapse (RPE) and the paired relapsed EC (P-RE) specimens. Microarray and RNA-seq were used to detect gene expression of EC samples. Clinicopathological characteristics and molecular data were compared between PE and P-M groups and between RPE and P-RE groups to explore the intra-tumor heterogeneity of EC. RESULTS:The clinical intra-tumor spatial heterogeneity of pathological type, grade, ER status, and PR status between PE and P-M were 17.9%, 13.1%, 28.6%, and 28.6%, respectively. The clinical intra-tumor spatiotemporal heterogeneity of pathological type, grade, ER status, and PR status between RPE and P-RE were 16.7%, 33.3%, 25.0%, and 37.5%, respectively. Cluster analysis sorts EC samples based on progression type of lesion and their pathological type. There were differentially expressed genes between PE and P-M and between RPE and P-RE, of which gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis were mainly enriched in cell proliferation, the p53 signaling pathway, etc. CONCLUSIONS:: Clinical and molecular data showed that there was spatiotemporal heterogeneity in intra-tumor of EC, which may add to the complexity of diagnosis and therapeutics for EC. Considering the intra-tumor heterogeneity, sequential chemotherapy and precision medicine may be a more suitable treatment plan for EC.

目的 应用RNAi技术干扰大鼠A7r5血管平滑肌细胞(vascular smooth muscle cells,VSMCs) Per2基因并检测VSMCs时钟基因Bmall、输出基因Dbp及AT1受体基因(AGTR1) mRNA表达的变化,初步探讨时钟基因、输出基因及AGTR1间的相关性.方法 应用已筛选携带大鼠Per2基因干扰序列的慢病毒载体转染大鼠血管平滑肌A7r5细胞,通过RT-PCR方法检测时钟基因Per2及Bmall、钟控基因Dbp和AGTR1 mRNA水平及节律变化.结果 慢病毒载体转染A7r5细胞后,RT-PCR方法检测钟基因Per2的mRNA峰值时相后移,表达节律异常;钟基因Bmall的mRNA表达峰值增强,表达节律未受影响;下游输出基因Dbp的mRNA表达明显受抑,表达节律异常;AGTR1的mRNA表达后期明显下降,提示其变化延迟于Per2变化.结论 通过RNAi技术沉默大鼠血管平滑肌细胞中时钟基因Per2表达,使钟基因Bmall反向增强,明显抑制输出基因Dbp的表达,影响其节律,可能造成AGTR1的异常延后表达.

It has been demonstrated that β‑arrestin2 is involved in the initiation and development of many types of cancers. However, its role in colorectal cancer (CRC) remains poorly understood. The present study investigated the role of β‑arrestin2 in CRC using CRC patient tissues as well as the LoVo and HCT116 CRC cell lines. Briefly, significantly higher expression of β‑arrestin2 was observed in CRC tissues compared with normal colon tissues. In addition, the downregulation of β‑arrestin2 reduced 5‑FU‑induced apoptosis in the LoVo cells, while the overexpression of β‑arrestin2 increased the apoptosis of HCT116 cells in vitro. Furthermore, the downregulation of β‑arrestin2 reduced the expression of the pro‑apoptotic proteins cleaved‑caspase‑3 and Bax, and increased the expression of the anti‑apoptotic protein Bcl‑2 after 5‑FU treatment. In addition, the expression of p‑p65 was increased after the β‑arrestin2 downregulation and was decreased after the β‑arrestin2 overexpression. However, β‑arrestin2 downregulation had no effect on the proliferation, migration and invasion capacity of the LoVo cells. In conclusion, these results indicated that β‑arrestin2 promoted 5‑FU‑induced CRC cell apoptosis via the NF‑κB pathway and may be used as a prognosis marker for CRC.

OBJECTIVE:CD4+CD25+ regulatory T cells (Tregs) have been found to have a decreased effector function in patients with multiple sclerosis (MS). In this study, we co-cultured naïve CD4+ T cells of MS patients with myelin basic protein (MBP)85-99 peptide as specific antigen and allogenic B cells as antigen-presenting cells, in an attempt to generate adequate antigen-specific CD4+CD25+ Tregs with normal or improved immune function.PATIENTS AND METHODS:Naïve CD4+ T cells were isolated from peripheral blood mononuclear cells (PBMCs) from patients with MS (n=5) and healthy controls (HC, n=5). Furthermore, these purified naive CD4+ T cells were co-cultured with the CD40-activated B cells and MBP85-99 peptide to induce MBP-reactive CD4+CD25highCD127low Tregs. After harvesting these Tregs via a flow sorter, real-time PCR and mixed lymphocyte reaction (MLR) assay were performed to characterize cellular immune function. Supernatant interleukin (IL)-10 and transforming growth factor (TGF)-β1 protein levels were detected by an enzyme-linked immunosorbent assay (ELISA).RESULTS:With this method, the frequency of CD4+CD25highCD127low Tregs in CD4+ T cells was 3.5%-6%. In both MS and HC groups, there were relatively lower proliferation indices (PI) of MLR assay but higher supernatant IL-10 and TGF-β1 levels in the presence of MBP than those in the presence of other control antigens, where no significant differences were found.CONCLUSIONS:Via the ex vivo culture, adequate MBP-reactive CD4+CD25+ Tregsderived from autologous naïve CD4+ T cells of MS patients, were obtained and returned to normal without immune defects, and even upregulated their immunosuppressive function mostly through the elevated release of IL-10 and TGF-β1.

FAM3A plays important roles in regulating hepatic glucose/lipid metabolism and the proliferation of VSMCs. This study determined the role and mechanism of FAM3A in the adipogenesis of 3T3-L1 preadipocytes. During the adipogenesis of 3T3-L1 preadipocytes, FAM3A expression was significantly increased. FAM3A overexpression enhanced 3T3-L1 preadipocyte adipogenesis with increased phosphorylated Akt (pAkt) level, whereas FAM3A silencing inhibited 3T3-L1 preadipocyte adipogenesis with reduced pAkt level. Moreover, FAM3A silencing reduced the expression and secretion of adipokines in 3T3-L1 cells. FAM3A protein is mainly located in mitochondrial fraction of 3T3-L1 cells and mouse adipose tissue. FAM3A overexpression increased, whereas FAM3A silencing decreased ATP production in 3T3-L1 preadipocytes. FAM3A-induced adipogenesis of 3T3-L1 preadipocytes was blunted by inhibitor of P2 receptor. In white adipose tissues of db/db and HFD-fed obese mice, FAM3A expression was reduced. One-month rosiglitazone administration upregulated FAM3A expression, and increased cellular ATP content and pAkt level in white adipose tissues of normal and obese mice. In conclusion, FAM3A enhances the adipogenesis of preadipocytes by activating ATP-P2 receptor-Akt pathway. Under obese condition, a decrease in FAM3A expression in adipose tissues plays important roles in the development of adipose dysfunction and type 2 diabetes.

Multiple sclerosis (MS) and neuromyelitis optica (NMO) are inflammatory demyelinating disorders of the central nervous system (CNS). Various genetic and environmental factors have been identified to contribute to etiology of MS and NMO. Aquaporin 4 (AQP4), is the most abundant water channel in CNS. AQP4 is expressed in astrocytes of the brain, spinal cord, optic nerve and supportive cells in sensory organs. In contrast to MS, immunoreactivity of AQP4 is abolished in NMO lesions. However, conflicting results have been reported regarding the association between AQP4 polymorphisms and demyelinating disorders. Considering the ethnic differences of genetic variations, replications in other cohorts are required. In this study, single nucleotide polymorphisms (SNPs) of AQP4 gene in patients with NMO/neuromyelitis optica spectrum disorders (NMOSD), and MS in the Northern Han Chinese population were examined. Six selected AQP4 SNPs were genotyped by high-resolution melting (HRM) method. Compared with healthy control (HC), there was no significant difference of AQP4 allele and genotype frequency in MS or NMO/NMOSD group. This study showed no significant association of common AQP4 SNPs with MS or NMO/NMOSD, strongly suggesting that polymorphisms of AQP4 gene are unlikely to confer MS or NMO/NMOSD susceptibility, at least in Northern Han Chinese population.

Background: Survivin is an oncoprotein silenced in normal mature tissues but reactivated in serous ovarian cancer (SOC). Although transcriptional activation is assumed for its overexpression, the long 3'-untranslated region (3'-UTR) in survivin gene, which contains many alternate polyadenylation (APA) sites, implies a propensity for posttranscriptional control and therefore was the aim of our study. Methods: The abundance of the coding region, the proximal and the distal region of survivin mRNA 3'-UTR, was evaluated by real-time polymerase chain reaction (PCR) in SOC samples, cell lines, and normal fallopian tube (NFT) tissues. The APA sites were confirmed by rapid amplification of cDNA 3' ends and DNA sequencing. Real-time PCR were used to screen survivin-targeting microRNAs (miRNAs) that were inversely correlated with survivin. The expression of an inversely correlated miRNA was restored by pre-miRNA transfection or induction with a genotoxic agent to test its inhibitory effect on survivin overexpression. Results: Varying degrees of APA were observed in SOC by comparing the abundance of the proximal and the distal region of survivin 3'-UTR, and changes of 3'-UTR correlated significantly with survivin expression (r = 0.708, P < 0.01). The main APA sites are proved at 1197 and 1673 of survivin 3'-UTR by DNA sequencing. Higher level of 3'-UTR proximal region than coding region was observed in NFT, as well as in SOC and cell lines. Among the survivin-targeting miRNAs, only a few highly expressed miRNAs were inversely correlated with survivin levels, and they mainly targeted the distal part of the 3'-UTR. However, in ovarian cancer cells, restoration of an inversely correlated miRNA (miR-34c) showed little effect on survivin expression. Conclusions: In NFT tissues, survivin is not transcriptionally silenced but regulate posttranscriptionally. In SOC, aberrant APA leads to the shortening of survivin 3'-UTR which enables it to escape the negative regulation of miRNAs and is responsible for survivin up-regulation.

OBJECTIVE:To investigate the role of myosin light chain kinase (MLCK) in intestinal barrier function in a mouse model with nonalcoholic steatohepatitis (NASH).METHODS:The C57BL/6 mice were randomly divided into five groups including control group, nonalcoholic fatty liver (NAFL) group, NAFL administrated with MLCK inhibitor ML-7 group, nonalcoholic steatohepatitis (NASH) group, NASH administrated with ML-7 group. Plasma ALT and AST were tested. The degree of liver steatosis was assessed by hematoxylin-eosin staining on liver tissue sections.Intestinal mucosal tight junction was observed by electron microscope. The expression of MLCK on intestinal mucosa was detected by immunohistochemistry staining. The level of lipopolysaccharide (LPS) in portal vein was determined by enzyme linked immune sorbent assay (ELISA). The protein and mRNA expression of inflammatory cytokines in liver tissue were tested using ELISA and real-time PCR.RESULTS:MLCK expression in intestinal mucosa was increased in NASH group compared with control group (P<0.01). The tight junctions of intestinal barrier were disrupted in NASH group and intercellular space was larger than control group [(26.60 ± 1.20) nm vs (14.90 ± 0.33) nm, P<0.05], which were improved after ML-7 administration [(14.9 0 ± 0.67) nm]. The LPS in portal vein was higher in NASH group than control group [(7.260 ± 3.184) U/L vs (2.962 ± 0.845) U/L, P<0.05], suggesting that the permeability of intestinal barrier was impaired, however the level of LPS was reduced by ML-7 [(3.772 ± 1.033) U/L, P<0.05]. ALT and AST in plasma, TNFα and IL-6 in liver tissue, the mRNA levels of TNFα and NF-κB in liver tissue were all elevated in NASH group compared with control group (all P<0.05), which were reduced by MLCK inhibitor ML-7.CONCLUSION:Epithelia MLCK probably plays a role in intestinal barrier impairment, which is critical to the pathogenesis of NASH.

Objective To investigate the hyperthermia-induced heat shock protein 70 (HSP70) expression and its significance in bladder carcinoma cell line EJ. Methods The 106/ml EJ was placed in 1.5 ml Eppendorf tube for water bath of different temperatures, which were induced by hyperthermia for 2 h, 4 h, 6 h, and 8 h, and recovered in 5% CO2 incubator (37 ℃) for 2 h. The flow cytometry (FCM) and Western blot were used to determine HSP70. RT-PCR was used to determine HSP70 mRNA. The SPSS16.0 statistical software was used to process the data. Results The HSP70 expression in human bladder carcinoma cell line EJ was increased along with the increasing temperature, with significant difference (P<0.05). However, the cell mortality was increased when the temperature exceeded 43 ℃. The HSP70 expression was increased along with the increasing induction time, with significant difference ( P<0.05). The HSP70 expression no longer increased at 8 h. Conclusion The optimal condition of hyperthermia-induced HSP70 expression in human bladder carcinoma cell line EJ is 43 ℃ for 6 h, which provides a certain experimental foundation for the production of tumor vaccine based on HSP70 bladder carcinoma.

After carefully checking the original data of migration and invasion experiment, we find we had selected the wrong picture to show the effect of microRNA320a on the migration of LoVo cells. The corrected version of Fig. 6A shown below contains appropriate representative images. We apologize for the error and appreciate the opportunity to correct the scientific record. All authors agree with this correction. [the original article was published in the Oncology Reports 27: 685-694, 2012 DOI: 10.3892/or.2011.1561]

本研究联合使用N-丁基-N-4-羟丁基亚硝胺(BBN)和抗坏血酸钠(Na-AsA)诱导小鼠膀胱肿瘤模型,观察热休克蛋白(HSP)70在膀胱癌发生发展中的表达及其与肿瘤微血管密度(MVD)的关系. 1 材料与方法 1.1 建立小鼠膀胱癌模型:6周龄纯系C57BL/6雄性小鼠60只,分笼饲养,每笼5只,室温24 ℃,相对湿度60%,按昼夜节律调节光照.

Objective To explore the association of common polymorphisms of aquaporin-4 (AQP-4) gene with multiple sclerosis (MS),neuromyelitis optica (NMO) in Chinese Northern Han population.Methods We enrolled patients diagnosed with MS (n =93),NMO (n =32),neuromyelitis optica spectrum disorders (NMOSD,n =35),clinically isolated syndrome (CIS,n =14) in Peking University People's Hospital,Beijing Tiantan Hospital and Beijing Fuxing Hospital from 2002 to 2013 and healthy controls (HC,n =109) who were healthy medical workers in Peking University People's Hospital from 2011 to 2013.Six chosen AQP-4 single-nucleotide polymorphisms (SNPs) were genotyped by highresolution melting method.Chi-square test was adopted to compare the frequency of alleles and genotypes of SNPs between MS,NMO,NMOSD,CIS patients and HC,as well as Bonferroni's method for adjustment analysis.Furthermore,stratification analysis was performed in terms of serum AQP-4 antibody in NMO group.Results Compared with HC,the frequency of A allele of rs162007 was nominally lower in MS (36.6％ (68/186) vs 47.6％ (98/206),odds ratio(OR) =0.635,95％ confidence interval(CI) 0.424-0.952,P =0.028) and NMOSD (33.8％ (23/68) vs 47.6％ (98/206) ; OR =0.563,95％ CI 0.318-0.998,P =0.048),while the frequency of T allele of rs3763043 was higher in CIS group (57.1％ (16/28)vs 37.0％ (74/200) ; OR =2.270,95％ CI 1.018-5.061,P =0.041) than that in HC.However,none of these SNPs was associated with MS,NMO,NMOSD or CIS after correction with Bonferroni's method.The stratification analysis also showed no remarkable distinction between NMO patients with positive and negative AQP-4 antibody.Conclusions No significant association of common AQP-4 gene polymorphisms with MS and NMO is found in Chinese Northern Han population,as well as with NMO positive for AQP-4 antibody,suggesting that the common polymorphisms of AQP-4 gene may not be associated with genetic susceptibility of these above-mentioned disorders including NMO.