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Differentiation induction subtraction hybridization (DISH): a strategy for cloning genes displaying differential expression during growth arrest and terminal differentiation.

Human cancers often display aberrant patterns of differentiation. By appropriate chemical manipulation, specific human cancers, such as human melanoma, leukemia and neuroblastoma, can be induced to lose growth potential irreversibly and terminally differentiate. Treatment of HO-1 human melanoma cells with a combination of recombinant human fibroblast interferon (IFN-beta) and the antileukemic compound mezerein (MEZ) results in irreversible growth arrest, a suppression in tumorigenic properties and terminal cell differentiation. A potential mechanism underlying these profound changes in cancer cell physiology is the activation of genes that can suppress the cancer phenotype and/or the inactivation of genes that promote the cancer state. To define the repertoire of genes modulated as a consequence of induction of growth arrest and terminal differentiation in human melanoma cells, we are using a differentiation induction subtraction hybridization (DISH) approach. A subtracted cDNA library, differentiation inducer treated cDNAs minus uninduced cDNAs, was constructed that uses temporally spaced mRNAs isolated from HO-1 cells treated with IFN-beta+MEZ. Approximately 400 random clones were isolated from the subtracted DISH library and analyzed by reverse Northern and Northern blotting approaches. These strategies resulted in the identification and cloning of both 30 known and 26 novel cDNAs displaying elevated expression in human melanoma cells induced to growth arrest and terminally differentiate by treatment with IFN-beta+MEZ. The DISH scheme and the genes presently identified using this approach should provide a framework for delineating the molecular basis of growth regulation, expression of the transformed phenotype and differentiation in melanoma and other cancers.

Antineoplastic Agents↗

Identification of differentially expressed genes with multivariate outlier analysis.

DNA microarray offers a powerful and effective technology to monitor the changes in the gene expression levels for thousands of genes simultaneously. It is being widely applied to explore the quantitative alternation in gene regulation in response to a variety of aspects including diseases and exposure of toxicant. A common task in analyzing microarray data is to identify the differentially expressed genes under two different experimental conditions. Because of the large number of genes and small number of arrays, and higher signal-noise ratio in microarray data, many traditional approaches seem improper. In this paper, a multivariate mixture model is applied to model the expression level of replicated arrays, considering the differentially expressed genes as the outliers of the expression data. In order to detect the outliers of the multivariate mixture model, an effective and robust statistical method is first applied to microarray analysis. This method is based on the analysis of kurtosis coefficient (KC) of the projected multivariate data arising from a mixture model so as to identify the outliers. We utilize the multivariate KC algorithm to our microarray experiment with the control and toxic treatment. After the processing of data, the differential genes are successfully identified from 1824 genes on the UCLA M07 microarray chip. We also use the RT-PCR method and two robust statistical methods, minimum covariance determinant (MCD) and minimum volume ellipsoid (MVE), to verify the expression level of outlier genes identified by KC algorithm. We conclude that the robust multivariate tool is practical and effective for the detection of differentially expressed genes.

Algorithms↗

Non-linear tests for identifying differentially expressed genes or genetic networks.

MOTIVATION: One of the recently developed statistics for identifying differentially expressed genetic networks is Hotelling T2 statistic, which is a quadratic form of difference in linear functions of means of gene expressions between two types of tissue samples, and so their power is limited. RESULTS: To improve the power of test statistics, a general statistical framework for construction of non-linear tests is presented, and two specific non-linear test statistics that use non-linear transformations of means are developed. Asymptotical distributions of the non-linear test statistics under the null and alternative hypothesis are derived. It has been proved that under some conditions the power of the non-linear test statistics is higher than that of the T2 statistic. Besides theory, to evaluate in practice the performance of the non-linear test statistics, they are applied to two real datasets. The preliminary results demonstrate that the P-values of the non-linear statistics for testing differential expressions of the genetic networks are much smaller than those of the T2 statistic. And furthermore simulations show the Type I errors of the non-linear statistics agree with the threshold used and the statistics fit the chi2 distribution. SUPPLEMENTARY INFORMATION: Supplementary data are available on Bioinformatics online.

Algorithms↗

Starch branching enzymes belonging to distinct enzyme families are differentially expressed during pea embryo development.

cDNA clones for two isoforms of starch branching enzyme (SBEI and SBEII) have been isolated from pea embryos and sequenced. The deduced amino acid sequences of pea SBEI and SBEII are closely related to starch branching enzymes of maize, rice, potato and cassava and a number of glycogen branching enzymes from yeast, mammals and several prokaryotic species. In comparison with SBEI, the deduced amino acid sequence of SBEII lacks a flexible domain at the N-terminus of the mature protein. This domain is also present in maize SBEII and rice SBEIII and resembles one previously reported for pea granule-bound starch synthase II (GBSSII). However, in each case it is missing from the other isoform of SBE from the same species. On the basis of this structural feature (which exists in some isoforms from both monocots and dicots) and other differences in sequence, SBEs from plants may be divided into two distinct enzyme families. There is strong evidence from our own and other work that the amylopectin products of the enzymes from these two families are qualitatively different. Pea SBEI and SBEII are differentially expressed during embryo development. SBEI is relatively highly expressed in young embryos whilst maximum expression of SBEII occurs in older embryos. The differential expression of isoforms which have distinct catalytic properties means that the contribution of each SBE isoform to starch biosynthesis changes during embryo development. Qualitative measurement of amylopectin from developing and maturing embryos confirms that the nature of amylopectin changes during pea embryo development and that this correlates with the differential expression of SBE isoforms.

1,4-alpha-Glucan Branching Enzyme↗

Differentially expressed genes execute zinc-induced apoptosis in precancerous esophageal epithelium of zinc-deficient rats.

Zinc deficiency (ZD) in rats increases esophageal cell proliferation and the incidence of N-nitrosomethylbenzylamine-induced esophageal tumors. Conversely, zinc replenishment (ZR) rapidly induces apoptosis in esophageal epithelia and reverses cancer development. We investigated gene expression changes in ZR versus ZD esophageal epithelia to identify differentially expressed genes associated with the antitumor effect of ZR. Weanling rats were fed a ZD diet for 6 weeks to establish esophageal cell proliferation or a zinc-sufficient (ZS) diet. Then, 10 ZD rats were treated with zinc gluconate intragastrically and switched to ZS diet; the remaining 10 ZD and ZS animals were treated with saline. All animals were killed 26-28 h later. Using cDNA microarrays, real-time polymerase chain reaction amplification and RNA hybridization techniques, we identified novel differentially expressed genes, including a RNA-binding protein with two RNA recognition motifs and a zinc knuckle (ZD7), and a DNA/RNA helicase with a DEAD box (ZD10) with two splice variants, ZD10a and ZD10b. In situ hybridization detected increased mRNA expression of ZD7, ZD10a and ZD10b in ZR esophageal epithelia, which displayed markedly increased occurrence of apoptotic cells, relative to ZD epithelia. Overexpression of ZD7 in human esophageal cancer cells resulted in induction of apoptosis and activation of caspase-3 and -7, activities that were inhibited by caspase-specific inhibitors. In addition, ZD7 mRNA levels and zinc-induced apoptosis in rat squamous carcinoma cells were reduced by specific small interfering ribonucleic acids. Thus, ZR rapidly induces ZD7 and ZD10 expression, which in turn stimulates apoptosis. These results provide the beginnings of a molecular pathway for zinc-induced apoptosis under conditions that reverse esophageal tumor initiation.

Animals↗

Differential expression of the L10 ribosomal protein during heart development.

Neural crest cells originating from the posterior rhombencephalon migrate to the cardiac outflow tract and participate in division of the aorta and pulmonary trunk. Ablation of this region of premigratory neural crest in the chick embryos results in nondivision of the outflow vessels. Subtractive hybridization was used to identify messages in the outflow tract of embryos with cardiac neural crest cells versus those lacking the cardiac neural crest cells. The chick L10 ribosomal protein was found to be differentially expressed in the outflow tract of embryos with cardiac neural crest. Several conditions have been identified that involve differential expression of ribosomal proteins, but this is the first report of differential expression in eukaryotic embryonic development.

Amino Acid Sequence↗

Identification of differentially expressed genes HSPC016 in dermal papilla cells with aggregative behavior.

The dermal papilla plays pivotal roles in hair follicle cycle and dermal papilla cells (DPCs) with aggregative behavior have more obviously inductive ability. In the present study, the suppression subtractive hybridization method was employed to identify the differentially expressed genes in dermal papillae cells with aggregative behavior. Following mRNA isolation of DPC with and without aggregative behavior, cDNA of both populations were prepared and subtracted by suppression PCR. Sequencing of enriched cDNAs identified five genes differentially expressed including capping protein, paladin, and vascular endothelial growth factor. Interestingly, HSPC016, first cloned from CD34+ hematopoietic stem/progenitor cells (HSPC), was identified by SSH, cDNA dot blot and Northern blot, which showed that this gene was differentially expressed in DPC with aggregative behavior. The full-length cDNA of HSPC016 was shown to be 366 bp, and the possible function of HSPC016 related to transcriptional regulation.

Amino Acid Sequence↗

Profiling of differentially expressed genes in human uterine leiomyomas.

Uterine leiomyomas are very common benign tumors resulting in clinically serious gynecological problems in women of reproductive age. Approximately, 1% of leiomyosarcoma was reported to arise in a preexisting leiomyoma. However, the molecular basis of these tumors is poorly understood. To understand the molecular changes during leiomyoma development, we profiled differentially expressed genes in ten paired leiomyoma and normal myometrial tissues using cDNA microarray chip analysis. We identified 67 genes (27 overexpressed and 40 underexpressed) which were scored as differentially expressed at least twofold in at least eight of ten patients. Eighteen of 67 genes have been already reported to be differentially expressed without their established functions in uterine leiomyoma and others have never been reported. Subsequently, the relative expression levels of representative genes from identified 67 genes were confirmed by reverse-transcriptase polymerase chain reaction and immunohistochemistry and were found to be consistent with the microarray data. This study could provide a new insight into the understanding of leiomyoma and leiomyosarcoma.

Adult↗

Differential expression of nitric oxide synthase isoforms in form-deprived chick eyes.

PURPOSE: To clarify whether nitric oxide synthase (NOS) is involved in development of myopia, we examined the influence of form deprivation on the expressions of NOS isoform mRNA. METHODS: NOS isoform cDNAs were amplified from total RNA extracted from control and 7-day-form-deprived chick retina-RPE (retinal pigment epithelium)-choroid, using competitive RT-PCR (reverse-transcription polymerase chain reaction). Each NOS isoform protein was also analyzed by Western blotting and immunohistochemistry. RESULT: Expression of inducible NOS (iNOS) mRNA was highest in the control chick retina-RPE-choroid, followed by the expression of brain NOS (bNOS) mRNA. Expression of endothelial NOS (eNOS) mRNA was faint. The iNOS protein level, however, was only slightly higher than the levels of the bNOS and eNOS proteins and was found mainly in the outer part of the photoreceptor layer and inner and outer parts of RPE and choroid. bNOS alone was found in the outer nuclear layer. Although form deprivation reduced the iNOS and bNOS mRNA expressions, only the iNOS protein showed significant reduction. CONCLUSION: All three NOS isoforms were expressed in chick retina-RPE-choroid. Predominant expression of iNOS, instead of bNOS and eNOS, suggested the existence of ocular tissue-specific regulation of the iNOS gene. In addition to differences in expression level, bNOS displayed regional differential expression. Moreover, only iNOS was reduced in response to form deprivation. It is suggested that NOS isoforms may be differentially involved in the mechanisms regulating the posterior eye tissues, including myopic eye growth.

Animals↗

Evidence for differential expression of CD45 isoforms by precursors for memory-dependent and independent cytotoxic responses: human CD8 memory CTLp selectively express CD45RO (UCHL1).

On the basis of differential CD45 expression, human T cells can be separated into approximately reciprocal populations. The mAb UCHL1 detects a 180 kd molecular mass isoform, CD45R0. The CD45RA cluster of mAbs reacts with CD45 isoforms of 205 and 220 kd molecular mass. These reagents subdivide both CD4 and CD8 T cell populations. CD4 T cells that proliferate in response to memory-dependent (recall) antigens have been shown to selectively express CD45R0. We extend these observations to a model for cytotoxic responses that allows the functional analysis of CD8 T cells with differential CD45 expression. Precursors for allo-specific CTL responses are readily detectable in CD45R0 as well as CD45RA populations. In contrast, we find memory CTLp greatly enriched among CD45R0 cells. In combination with earlier work, these results suggest that differential expression of CD45 isoforms is associated with memory formation for different classes of immune responses in both major T cell lineages.

Antigens, Differentiation↗

[Gene chip screen in allergic rhinitis and preliminary analysis of the differential expressed genes].

OBJECTIVE: To discover the etiology of allergic rhinitis(AR) at the genomic level. METHODS: Microarray analysis on a genomic scale was used to find changes in genes expression in AR. The total RNAs were isolated from nasal mucosa of AR and normal, then purified to mRNAs and were reversely transcribed to cDNAs with the incorporation of fluorescent-labeled dUTP to prepare for the hybridization probes. The genes differential expressed in AR were screened with the gene chip which contained 12,800 human genes in all 8 samples. RESULT: 734 genes were shown in differential expressed profile with 430 upregulated genes and 304 downregulated genes, and all 8 samples were 15 genes upregulated and 6 genes downregulated. The differential expressed genes involved in the functions of inflammation, immunology, neuroendocrine and signal conduct, and they had potential value in AR study. CONCLUSION: The investigation based on gene chip in researching related genes in AR afforded a new idea in studying pathogenesis of nasal allergic diseases.

DNA, Complementary↗

AffyMiner: mining differentially expressed genes and biological knowledge in GeneChip microarray data.

BACKGROUND: DNA microarrays are a powerful tool for monitoring the expression of tens of thousands of genes simultaneously. With the advance of microarray technology, the challenge issue becomes how to analyze a large amount of microarray data and make biological sense of them. Affymetrix GeneChips are widely used microarrays, where a variety of statistical algorithms have been explored and used for detecting significant genes in the experiment. These methods rely solely on the quantitative data, i.e., signal intensity; however, qualitative data are also important parameters in detecting differentially expressed genes. RESULTS: AffyMiner is a tool developed for detecting differentially expressed genes in Affymetrix GeneChip microarray data and for associating gene annotation and gene ontology information with the genes detected. AffyMiner consists of the functional modules, GeneFinder for detecting significant genes in a treatment versus control experiment and GOTree for mapping genes of interest onto the Gene Ontology (GO) space; and interfaces to run Cluster, a program for clustering analysis, and GenMAPP, a program for pathway analysis. AffyMiner has been used for analyzing the GeneChip data and the results were presented in several publications. CONCLUSION: AffyMiner fills an important gap in finding differentially expressed genes in Affymetrix GeneChip microarray data. AffyMiner effectively deals with multiple replicates in the experiment and takes into account both quantitative and qualitative data in identifying significant genes. AffyMiner reduces the time and effort needed to compare data from multiple arrays and to interpret the possible biological implications associated with significant changes in a gene's expression.

Algorithms↗

Identification of differentially expressed genes by serial analysis of gene expression in human prostate cancer.

Prostate cancer is the leading cause of cancer death in American males. To better understand the genetic bases of this disease, we have generated a comprehensive molecular profile of human prostate. The gene expression pattern in normal and prostate cancer tissues was analyzed by serial analysis of gene expression (SAGE). A total of 133,217 transcripts were analyzed, and 35,185 distinct SAGE tags were identified representing 19,287 genes. Comparison of the transcripts in normal and tumor tissue revealed 156 differentially expressed genes (P < 0.05), of which 88 genes were up-regulated and 68 genes were down-regulated in the tumor tissue. Based on SAGE data, we estimate that the transcriptome for human prostate is approximately 37,000. Several differentially expressed genes identified by SAGE were selected for confirmation using immunohistochemistry. Some genes (e.g., E2F4) were overexpressed in tumor epithelial cells and some (e.g., Daxx) were increased in tumor stroma. Further characterization of the role of E2F4 and Daxx as well as other differentially expressed genes may provide useful insights into the mechanism of prostate cancer development.

Dihydrotestosterone↗

Isolation, sequence analysis and expression profile of a novel porcine gene, NIP7, differentially expressed in the Longissimus dorsi muscle tissues from Meishan, Meishan x Large White cross and Large White pigs.

In order to detect the molecular mechanism of heterosis in pigs, the mRNA differential display technique was performed to investigate the differences in gene expression in the Longissimus dorsi muscle tissues from Meishan, Meishan x Large White cross and Large White pigs. One novel gene that was differentially expressed was identified using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and its complete cDNA sequence was obtained using the rapid amplification of cDNA ends (RACE) method. The nucleotide sequence of the gene is not homologous to any of the known porcine genes. The sequence prediction analysis revealed that the open reading frame of this gene encodes a protein of 180 amino acids that contains the conserved putative RNA-binding domain in PseudoUridine synthase and Archaeosine transglycosylase (PUA) and has high homology with the 60S ribosome subunit biogenesis protein NIP7 homolog of three species--human (98%), mouse (97%) and rat (96%)--so that it can be defined as swine 60S ribosome subunit biogenesis protein NIP7 homolog (NIP7). The tissue expression analysis indicated that the swine NIP7 gene is over expressed in muscle, heart, liver, fat, kidney, and lung, but weakly expressed in small intestine, ovary, and spleen. The genomic DNA sequence of swine NIP7 gene was finally amplified and result revealed that the swine NIP7 gene contains five exons and four introns. Our experiment is the first to establish the primary foundation for further research on the swine NIP7 gene.

Amino Acid Sequence↗

Microarray analysis of differentially expressed genes in vaginal tissues from women with stress urinary incontinence compared with asymptomatic women.

BACKGROUND: The pathophysiology of pelvic floor dysfunction resulting in stress urinary incontinence (SUI) in women is complex. Evidence suggests that there is also a genetic predisposition towards SUI. We sought to identify differentially expressed genes involved in extracellular matrix (ECM) metabolism in vaginal tissues from women with SUI in the secretory phase of menses compared with asymptomatic women. METHODS: Tissue samples were taken from the periurethral vaginal wall of five pairs of premenopausal, age-matched SUI and continent women and subjected to microarray analysis using the GeneChip Human Genome U133 oligonucleotide chip set. RESULTS: Extensive statistical analyses generated a list of 79 differentially expressed genes. Elafin, keratin 16, collagen type XVII and plakophilin 1 were consistently identified as up-regulated ECM genes. Elafin, a serine protease inhibitor involved in the elastin degradation pathway and wound healing, was expressed in pelvic fibroblasts and confirmed by Western blot, quantitative competitive PCR and immunofluorescence cell staining. CONCLUSIONS: Genes involved in elastin metabolism were differentially expressed in vaginal tissue from women with SUI, suggesting that elastin remodelling may be important in the molecular aetiology of SUI.

Adult↗

Differential expression of glutathione S-transferase isoforms in compartments of the testis and segments of the epididymis of the rat.

Specific cell types of the mammalian testes demonstrate varying susceptibility to toxic insult by chemical agents. The mammalian testis is divided into two major compartments: seminiferous tubules, the site of spermatogenesis, and interstitium, which contains the Leydig cells. Glutathione S-transferase (GST) expression was examined in isolated compartments of the rat testis and in segments of the epididymis. Western blot analysis revealed the presence of GST class alpha, mu, and pi bands in each of the isolated compartments of the testis, and HPLC analysis of monomeric isoforms provided evidence for differential expression of multiple GST isoforms in testicular compartments. All major isoforms (e.g., forms 1, 2, 3, 4, 6, 7, 8, 9, and 11) were detected in the cytosol of whole testis. Isoform subunit 4 was the major form in the tubule, whereas isoform subunit 11 is the dominant form in the Leydig cells. Isoform subunits 3, 4, and 6 were enriched in the tubules as compared to interstitial or Leydig cells. The preferential action of reproductive toxicants at specific stages of aging may be due to an age-dependent expression of the activating or detoxifying enzymes in the reproductive tract. Therefore, the age-dependent expression of testicular GST isoforms was also examined. Expression of isoform subunits 2 and 4 displayed an age dependence, with the largest increase in these subunits occurring between ages 4 and 15 weeks. Isoform expression did not correlate with serum testosterone levels. HPLC analysis of the GST isoforms in the longitudinal segments of the epididymis and vas deferens revealed differential expression within these segments. Total GST protein and catalytic activity was highest in the caput epididymis and progressively decreased toward the vas deferens. Isoform subunit 2 was the major form expressed in the epididymis. The results of this study indicate that the GSTs are differentially expressed in testicular compartments and epididymal segments, and that this may contribute to susceptibility of different cell types to xenobiotic damage.

Animals↗

Human myometrial genes are differentially expressed in labor: a suppression subtractive hybridization study.

Human parturition is effected by a cascade of factors, of which many are unknown. We aim to identify the genes that are changed by labor in the human myometrium by suppression subtractive hybridization. We also seek to ascertain whether these genes are differentially expressed in the myometrium at the upper or fundal and lower segments of the uterus. Term myometrial tissues were obtained from laboring and nonlaboring women undergoing cesarean section after obtaining informed consent. Total RNA was used in suppression subtractive hybridization (CLONTECH PCR Select) to produce two subtracted cDNA libraries enriched for genes expressed during or before labor, labor and not-in-labor libraries, respectively. Dot blot screening of 400 positive clones, constituting 20% of the two subtracted libraries, revealed 30 differentially expressed clones, 14 of which were up-regulated by labor. Among the 10 known genes that were up-regulated in labor, 6 had apparent immune regulatory and inflammatory roles. Three are well-known inflammatory mediators and modulators that were previously linked with parturition: IL-8, manganese superoxide dismutase (MnSOD), and metalloproteinase-9. Three others, interferon-inducible 1-8d gene, elongation factor 1alpha, and nucleophosmin, have not been previously linked with labor. Constitutively expressed genes, including cyclophilin and alpha-actin, were found to be altered by labor. Quantitative real-time RT-PCR using Taqman probes further confirmed the up-regulation of some of these genes. The amounts of the specific genes assayed were standardized to 18S ribosomal RNA and are expressed as mean +/- SEM. Quantitative real-time RT-PCR showed that IL-8 mRNA rose from 0.003 +/- 0.002 in nonlaboring samples (n = 38) to 0.24 +/- 0.11 (n = 20) in gestational-age-matched spontaneously laboring women (P = 0.035). Similarly, MnSOD rose from 0.11 +/- 0.02 (n = 24) to 1.23 +/- 0.56 (n = 24) in gestational-age-matched women (P = 0.047). Additionally, cyclophilin, often used as a constitutive or housekeeping gene marker, increased from 0.0008 +/- 0.0002 (n = 6) to 0.002 +/- 0.0004 (n = 6; P = 0.008) during labor. Notably, MnSOD mRNA was differentially distributed between the upper (0.63 +/- 0.18) and lower (0.15 +/- 0.05; n = 15; P = 0.022) segments of the uterus, but IL-8 was not (n = 17; P = 0.97). Induced labor further showed significantly higher levels of IL-8 (0.63 +/- 0.21; n = 14) than spontaneous labor (0.22 +/- 0.11; n = 20; P = 0.046), but not MnSOD (P = 0.1). This work identifies novel as well as known genes that were not previously associated with parturition. It extends previous data indicating that there is differential expression of some, but not all genes within the gravid human uterus. Inflammatory genes constitute a major proportion of the known genes found to be up-regulated in labor, lending support to the hypothesis of an inflammatory mechanism for human parturition. This work further indicates that many factors associated with human labor and their complex interactions remain to be elucidated.

Computer Systems↗

Differential expression of S100 gene family in human esophageal squamous cell carcinoma.

PURPOSE: To study the differential expression of the S100 gene family at the RNA level in human esophageal squamous cell carcinoma (ESCC), and to find the relationship of the S100 gene family with ESCC. METHODS: Firstly, the specific primers were designed for the different S100 genes with Software Primer 3, which required that both primer sequences of each S100 gene were from two different exons respectively. Then, the differential expression of 16 S100 genes was examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) in 62 cases of ESCC versus the corresponding normal esophageal mucosa. All RT-PCR products were analyzed by 1.5% agarose gel. With Fluor-S MultiImager and Multi-Analyst software, the electrophoresis images were evaluated with statistics analysis using SAS 8.1 software. RESULTS: Eleven out of 16 S100 genes were significantly downregulated ( p<0.05) in ESCC versus the normal counterparts such as S100A1, S100A2, S100A4, S100A8, S100A9, S100A10, S100A11, S100A12, S100A14, S100B, and S100P genes. Only the S100A7 gene in the S100 family was markedly upregulated ( p<0.05). Moreover, the S100B gene was significantly correlated with histological differentiation of ESCC ( p=0.0247), and the deregulation of some S100 genes was closely correlated ( p<0.05), such as S100A10/S100A11, S100A2/S100A8, S100A2/S100A14, S100A8/S100A14, and S100A2/S100P etc. CONCLUSIONS: The S100 gene family is closely associated with ESCC.

Carcinoma, Squamous Cell↗