CCR9 AND CCR7 are overexpressed in CD4- CD8- thymocytes of myasthenia gravis patients.

INTRODUCTION: Chemokine CC motif receptors 9 and 7 (CCR9 and CCR7) play a major role in the migration of T-cell precursors to the thymus to initiate T thymopoiesis. However, their role in development of T-cells in myasthenia gravis (MG) patients has not been fully elucidated. METHODS: Expression and distribution of CCR9+ and CCR7+ cells were detected by flow cytometry and immunofluorescence. Real-time polymerase chain reaction was used to check the adhesion molecules on CD4- CD8- double-negative (DN) thymocytes. RESULTS: CCR9 and CCR7 expression by DN thymocytes increased in the MG thymus; the levels of CCR9, CCR7, interleukin-7R mRNA increased, and CXCR4 levels decreased compared with levels in the non-MG thymus. More CCR7 and CCR9 double-positive (DP) thymocytes were gathered near the subcapsular region in MG thymus. CONCLUSIONS: Enhanced expression of CCR9 and CCR7 may complicate the differentiation of DP thymocytes from the DN stage in MG thymus. Muscle Nerve, 2016 Muscle Nerve 55: 84-90, 2017.

Dysfunction of the PGC-1α-mitochondria axis confers adriamycin-induced podocyte injury.

Adriamycin (ADR)-induced nephropathy in animals is an experimental analog of human focal segmental glomerulosclerosis, which presents as severe podocyte injury and massive proteinuria and has a poorly understood mechanism. The present study was designed to test the hypothesis that the peroxisome proliferator-activated receptor-γ coactivator (PGC)-1α-mitochondria axis is involved in ADR-induced podocyte injury. Using MPC5 immortalized mouse podocytes, ADR dose dependently induced downregulation of nephrin and podocin, cell apoptosis, and mitochondrial dysfunction based on the increase in mitochondrial ROS production, decrease in mitochondrial DNA copy number, and reduction of mitochondrial membrane potential and ATP content. Moreover, ADR treatment also remarkably reduced the expression of PGC-1α, an important regulator of mitochondrial biogenesis and function, in podocytes. Strikingly, PGC-1α overexpression markedly attenuated mitochondrial dysfunction, the reduction of nephrin and podocin, and the apoptotic response in podocytes after ADR treatment. Moreover, downregulation of PGC-1α and mitochondria disruption in podocytes were also observed in rat kidneys with ADR administration, suggesting that the PGC-1α-mitochondria axis is relevant to in vivo ADR-induced podocyte damage. Taken together, these novel findings suggest that dysfunction of the PGC-1α-mitochondria axis is highly involved in ADR-induced podocyte injury. Targeting PGC-1α may be a novel strategy for the treatment of ADR nephropathy and human focal segmental glomerulosclerosis.

Increased expression levels of S100A4 associated with hypoxia-induced invasion and metastasis in esophageal squamous cell cancer.

Here, we explored the expression of S100A4 in esophageal squamous cell cancer (ESCC) tissues and investigated its role in hypoxia-induced invasion and metastasis in ESCC cell lines EC-1 and EC-9706. Immunohistochemistry analysis demonstrated that S100A4 was overexpressed in human ESCC tissues especially in ESCC tissues with deep invasion and lymph node metastasis. Hypoxia-induced S100A4 overexpression was observed in EC-1 and EC-9706 cells, in which it was associated with invasion and metastasis. Furthermore, we used EC-1 and EC-9706 cells again to upregulate or knockdown the expression S100A4 to investigate the mechanism role of S100A4 in hypoxia-induced invasion and metastasis in ESCC cells. And the results showed that S100A4 played an important role in promoting the invasion and metastasis of EC-1 and EC-9706 cells under hypoxia. Therefore, S100A4 overexpression might be an important mechanism by which hypoxia induced invasion and metastasis, and S100A4 could also be a potential target for the treatment of ESCC.

Notch-1-mediated esophageal carcinoma EC-9706 cell invasion and metastasis by inducing epithelial-mesenchymal transition through Snail.

Notch has recently been shown to promote epithelial-to-mesenchymal transition (EMT) by involving in the EMT process that occurs during tumor progression and converts polarized epithelial cells into motile, invasive cells. However, it is still unclear whether the Notch signaling pathway is associated with the regulation of EMT in esophageal carcinoma. The present study explored Notch-1-mediated esophageal carcinoma EC-9706 cell invasion and metastasis by inducing epithelial­mesenchymal transition through Snail. The results demonstrated that the inhibition of Notch-1 expression in the esophageal carcinoma cell line EC-9706 could suppress the occurrence of EMT and at the same time could decrease the invasion and metastasis ability of the EC-9706 cells, indicative of its role in EMT. Snail is a transcriptional repressor of E-cadherin. We found that with the inhibition of Notch-1 expression in the esophageal carcinoma cell line EC-9706, the expression of Snail also decreased. Mechanistic studies showed that the up-expression of Snail in the EC-9706 cells restored the suppression of EMT regulated by Notch-1 inhibition, suggesting the role of Snail in Notch-1-mediated EMT. At the same time, the up-expression of Snail in the EC-9706 cells could also rescue the invasion and metastasis ability inhibited by Notch-1 siRNA. Taken together, our results had revealed that Notch-1 could participate in the invasion and metastasis of esophageal carcinoma through EMT via Snail. This study indicated that Notch-1 might be a useful target for esophageal carcinoma prevention and therapy.

Differential expression profiling of microRNAs and their potential involvement in esophageal squamous cell carcinoma.

MicroRNAs are small, noncoding RNAs approximately 18-24 nucleotides in length that negatively regulate gene expression at the posttranscriptional and/or translational level by binding to complimentary sequences in the 3'-untranslated regions of target mRNAs. Growing evidence has indicated the important roles for different miRNA species in the development of different cancers. Therefore, miRNAs have the potential to become new biological markers for esophageal squamous cell carcinoma (ESCC) and to be applied in the diagnosis, prognosis, and targeted treatment of ESCC. In this study, we performed a miRNA microarray to analyze the miRNA expression profile in ESCC compared to normal tissues. Then, we made a preliminary analysis of the biological function for the most differentially expressed miRNAs and their potentially target genes regulated. Some microarray results were validated by performing quantitative RT-PCR. The study provided evidence that linked the biological role of miRNAs to ESCC and showed that miRNAs could undertake a variety of mechanisms. Additionally, we also found that altered miR-429 and miR-451 expression levels were associated with the occurrence of lymph node metastases and the differentiation status and TNM stage in ESCC. The study of miRNAs may lead to finding novel methods to diagnose, treat, and prevent ESCC.

DNA polymerase ß mutations and survival of patients with esophageal squamous cell carcinoma in Linzhou City, China.

Linzhou City in northern China has a high incidence of esophageal squamous cell carcinoma (ESCC). This study retrospectively analyzed the data of 231 cases with ESCC collected from 1998 to 2012. Mutations of DNA polymerase ß (polß) gene in the ESCC samples from patients in Linzhou City were examined by amplifying polß cDNA by RT-PCR followed by cloning and sequencing. Mutations in polß were found in 105 cases (45.9%). Nine types of mutations were identified in the polß cDNA; the most common were 177­234 nt deletion (11.3%), 462 nt G → T (9.1%), and 648 nt G → C (6.9%). Mutations in polß appeared to be associated with TNM status (P = 0.048). Follow-up data was used for survival analysis. The overall 5-year survival rate of the 231 patients was 37.4%; the rate for patients with wild-type (WT) polß was 41.8%. Compared with the WT polß group, the median survival for patients with specific mutations (177­234 nt deletion, 462 nt G → T, or 613 nt A → T) was significantly shorter (all P = 0.000), and the 5-year survival rate decreased to 0%. Patients with the 648 nt G → C mutation had improved survival (P = 0.000) with a 5-year survival rate of 100%. Our results identified nine types of mutations within polß cDNA in ESCC patients with four mutations related to patient survival.

Myricetin exerts anti-proliferative, anti-invasive, and pro-apoptotic effects on esophageal carcinoma EC9706 and KYSE30 cells via RSK2.

Myricetin, a common dietary flavonoid, is widely distributed in fruits and vegetables and is used as a health food supplement based on its anti-tumor properties. However, the effect and mechanisms of myricetin in esophageal carcinoma are not fully understood. Here, we demonstrated the effect of myricetin on the proliferation, apoptosis, and invasion of the esophageal carcinoma cell lines EC9706 and KYSE30 and explored the underlying mechanism and target protein(s) of myricetin. CCK-8 assay, transwell invasion assay, wound-healing assay, cell cycle analysis, and apoptosis assay were used to evaluate the effects of myricetin on cell proliferation, invasion, and apoptosis. Nude mouse tumor xenograft model was built to understand the interaction between myricetin and NTD RSK2. Pull-down assay was used to verify molecular mechanism. Myricetin inhibited proliferation and invasion and induced apoptosis of EC9706 and KYSE30 cells. Moreover, myricetin was shown to bind RSK2 through the NH2-terminal kinase domain. Finally, myricetin inhibited EC9706 and KYSE30 cell proliferation through Mad1 and induced cell apoptosis via Bad. Myricetin inhibits the proliferation and invasion and induces apoptosis in EC9706 and KYSE30 cells via RSK2. Myricetin exerts anti-proliferative, anti-invasive, and pro-apoptotic effects on esophageal carcinoma EC9706 and KYSE30 cells via RSK2. Our results provide novel insight into myricetin as a potential agent for the prevention and treatment of esophageal carcinoma.

Umbilical cord blood-derived dendritic cells loaded with BGC823 tumor antigens and DC-derived exosomes stimulate efficient cytotoxic T-lymphocyte responses and antitumor immunity in vitro and in vivo.

BACKGROUND: Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells and from which a significant number of dendritic cells (DCs) can be produced. But the therapeutic role of DCs and exosomes (EXO) generated from DCs is not fully elucidated. MATERIAL AND METHODS: The UCB-derived DCs were loaded with tumor antigens generated from BGC823 cell line. Exosomes were derived from these DCs by ultracentrifugation. Dendritic cells and DCex were evaluated by light microscope, transmission electron microscope (TEM), flow cytometry, and western blot assay. The therapeutic role of DCs and EXO generated from DCs were then detected in vitro and in vivo. RESULTS: Dendritic cells isolated from umbilical cord blood after loading with tumor antigens generated from BGC823 cell line could express high levels of protein molecules: MHC-I, MHC-II, CD34, CD40, CD80, CD86, CD11c and CD54 and mediate a stronger promotion of T cells proliferation. And, they could also enhance the cytotoxicity effects of the generated CTL in vitro and in vivo. Exosomes isolated from these DCs were 40-90-nm round particles with a complete membrane structure and could also expressed molecules similar to DCs. Exosomes could stimulate T cell proliferation, produce effective cytotoxicity and induce more efficient in vivo antitumor immunity. CONCLUSIONS: These results suggested that tumor antigens loaded DCs derived from unrelated umbilical cord blood or DCex can induce tumor specific cytotoxicity and this may represent a novel immunotherapy for tumors. Because of their advantage of stable, easy to store, DCex have a more brilliant prospects in the tumor immunity. ADDITIONAL INFORMATION: We reported that exosomes derived from umbilical cord blood dendritic cell (UBDC), similar to DCs, can trigger activation of T cells significantly. These data demonstrate that DC-derived exosomes (DCex) can mediate essential adaptive immune functions.

DNA polymerase beta promoter mutations affect gene transcription, translation and the sensitivity of esophageal cancer cells to cisplatin treatment.

The ability of a promoter to initiate transcription is important for the control of gene expression. Mutations in the DNA polymerase beta (po1ß) promoter may affect the transcription of this gene; however, the relationship between these mutations and the upregulation of the expression of po1ß remains unclear. Therefore, in the present study, three po1ß promoter mutants (M1, -37 C→A; M2, -114 G→A, -37 C→A; M3, -194 T→C) were generated to examine the effect of promoter mutations on polß gene expression and sensitivity to cisplatin. We found that the M1 and M2 mutant polß promoter constructs showed higher RLA than the wild-type polß promoter (P < 0.01), whereas the activity of the M3 polß promoter did not differ significantly from that of the wild-type polß promoter (P > 0.05). The expression levels of polß mRNA and protein were significantly higher (P < 0.01) and the sensitivity to cisplatin was significantly lower (P < 0.05) in Eca9706(-/-)-M1 and Eca9706(-/-)-M2 cells than in Eca9706(-/-)-W. The expression levels of polß mRNA and protein and the sensitivity to cisplatin were not significantly different between Eca9706(-/-)-M3 and Eca9706(-/-)-W cells (P > 0.05).These results revealed that specific mutations of the polymerase beta gene promoter significantly enhanced the gene's transcriptional activity. These mutations correspondingly increased the gene's mRNA and protein product, at the same time reduced the esophageal cancer cells' sensitivity to cisplatin.

[Retracted] miRNA­1207­5p is associated with cancer progression by targeting stomatin­like protein 2 in esophageal carcinoma.

Subsequently to the publication of the above article, and a Corrigendum that has already been published with the intention of showing corrected versions of Figs. 3 and 6 (DOI: 10.3892/ijo.2018.4254; published online on January 24, 2018), a concerned reader drew to the Editor's attention that there appeared to be an unexpected overlap of data in a couple of the panels showing flow cytometric data in Fig. 3A; furthermore, strikingly similar data also appeared in a paper that was submitted to the journal Cancer Gene Therapy at around the same time [Zang W, Wang T, Huang J, Li M, Wang Y, Du Y, Chen X and Zhao G: Long noncoding RNA PEG10 regulates proliferation and invasion of esophageal cancer cells. Cancer Gene Ther 22: 138­144, 2015]. Considering the latest discrepancies and concerns that have been raised with another of the figures in this paper, the Editor of International Journal of Oncology has decided that the article should be retracted from the publication. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership of the Journal for any inconvenience caused. [International Journal of Oncology 46: 2163­2171, 2015; DOI: 10.3892/ijo.2015.2900].

Ring1a protects against colitis through regulating mucosal immune system and colonic microbial ecology.

Inflammatory bowel disease (IBD) represents a prominent chronic immune-mediated inflammatory disorder, yet its etiology remains poorly comprehended, encompassing intricate interactions between genetics, immunity, and the gut microbiome. This study uncovers a novel colitis-associated risk gene, namely Ring1a, which regulates the mucosal immune response and intestinal microbiota. Ring1a deficiency exacerbates colitis by impairing the immune system. Concomitantly, Ring1a deficiency led to a Prevotella genus-dominated pathogenic microenvironment, which can be horizontally transmitted to co-housed wild type (WT) mice, consequently intensifying dextran sodium sulfate (DSS)-induced colitis. Furthermore, we identified a potential mechanism linking the altered microbiota in Ring1aKO mice to decreased levels of IgA, and we demonstrated that metronidazole administration could ameliorate colitis progression in Ring1aKO mice, likely by reducing the abundance of the Prevotella genus. We also elucidated the immune landscape of DSS colitis and revealed the disruption of intestinal immune homeostasis associated with Ring1a deficiency. Collectively, these findings highlight Ring1a as a prospective candidate risk gene for colitis and suggest metronidazole as a potential therapeutic option for clinically managing Prevotella genus-dominated colitis.

We found that PcG protein Ring1a could be a new risk gene for colitis. Ring1a deficiency causes aggravated colitis by regulating the mucosal immune system and colonic microbial ecology.

Relationship between TWIST expression and epithelial-mesenchymal transition of oesophageal squamous cell carcinoma.

We have investigated mRNA and protein expression of TWIST, Vimentin and E-cadherin in ESCC (oesophageal squamous cell carcinoma) and explored their relationship with tumour's infiltration and metastasis. RT-PCR (reverse transcriptase-PCR) was used to evaluate mRNA expression of TWIST, E-cadherin and Vimentin in 40 cases of ESCC. The protein expression of the genes was examined by immunohistochemical staining in each specimen. Expression of TWIST, E-cadherin and Vimentin mRNA and protein with clinicopathologic parameters were analysed. mRNAs of TWIST, Vimentin and E-cadherin were expressed in 75, 55 and 35% respectively of ESCC, i.e. significantly different from that in normal oesophageal mucosa (15, 0 and 85% respectively; P<0.01). In ESCC with LN (lymph node) metastasis, expression of TWIST and Vimentin mRNA, but not E-cadherin mRNA was significantly higher (100 and 83%) than in ESCC without LN metastasis (64 and 43%, P=0.018) respectively. Levels of mRNA expression of the 3 genes followed similar patterns to their above-mentioned frequencies. Protein expression of TWIST, E-cadherin and Vimentin were observed in 70, 35 and 50% respectively of ESCC, which were significantly different from normal mucosa (15, 80 and 0%; P<0.001). In ESCC with LN metastasis, protein expression of TWIST and Vimentin, but not E-cadherin, were significantly higher (100 and 75%) than in ESCC without LN metastasis (61 and 39%). Protein expression of TWIST was positively correlated with Vimentin (r=0.327, P=0.039), but negatively correlated with E-cadherin (r= -0.633, P=0.000). Thus, both mRNAs and proteins of TWIST and Vimentin were significantly overexpressed in ESCC, especially ESCC with LN metastasis. The mRNA and protein of E-cadherin were down-regulated in ESCC. These results suggest potential roles of TWIST as the promoter of tumour invasion and metastasis associated with down-regulation of E-cadherin.

S100A4 mediated cell invasion and metastasis of esophageal squamous cell carcinoma via the regulation of MMP-2 and E-cadherin activity.

It is well documented that S100A4 is upregulated in a large amount of invasive tumors and plays a pivotal role in tumor invasion and metastasis. However, the precise role and mechanism S100A4 exerts in the invasion and metastasis of esophageal squamous cell carcinoma (ESCC) have not been fully elucidated to date. Our data demonstrated that S100A4 was overexpressed in human ESCC tissues, especially in ESCC with poor differentiation, deep invasion and lymph node metastasis. Subsequently, the knockdown of S100A4 by RNAi in ESCC cell line (EC-1) could reduce cell invasion, metastasis and proliferation ability in vitro. Most importantly, S100A4 regulated MMP-2 positively and E-cadherin negatively in vivo and in vitro to some extent. Our results suggest that S100A4 is an important factor in the invasion, metastasis and proliferation of ESCC and may control invasion and metastasis at least in part through the regulation of MMP-2 and E-cadherin activity. S100A4 may serve as a biomarker for progression of ESCC and a potential molecular target for biotherapy of ESCC.

New Approach to Intelligence Screening for Children With Global Development Delay Using Eye-Tracking Technology: A Pilot Study.

Objective: There has become a consensus for detecting intellectual disability in its early stages and implementing effective intervention. However, there are many difficulties and limitations in the evaluation of intelligence-related scales in low-age children. Eye-tracking technology may effectively solve some of the pain points in the evaluation. Method: We used an eye-tracking technology for cognitive assessment. The subjects looked at a series of task pictures and short videos, the fixation points of which were recorded by the eye-movement analyzer, and the data were statistically analyzed. A total of 120 children aged between 1.5 and 4 years participated in the study, including 60 typically developing children and 60 children with global development delay, all of whom were assessed via the Bayley scale, Peabody Picture Vocabulary Test (PPVT), and Gesell scale. Results: Cognitive scores from eye-tracking technology are closely related to the scores of neuropsychological tests, which shows that the technique performs well as an early diagnostic test of children's intelligence. Conclusions: The results show that children's cognitive development can be quickly screened using eye-tracking technology and that it can track quantitative intelligence scores and sensitively detect intellectual impairment.

Exosomes derived from LPS-stimulated human thymic mesenchymal stromal cells enhance inflammation via thrombospondin-1.

Inflammatory response mediated by immune cells is either directly or indirectly regulated by mesenchymal stromal cells (MSCs). Accumulating evidence suggests that thrombospondin-1 (TSP-1) is highly expressed in response to inflammation. In this work, we isolated and identified human thymic mesenchymal stromal cells (tMSCs) and detected the expression of TSP-1. We found that tMSCs expressed TSP-1 and Poly (I:C) or LPS treatment promoted the expression of TSP-1. Further, we isolated and identified exosomes originating from tMSCs (MEXs). Notably, exosomes derived from LPS-pretreated tMSCs (MEXsLPS) promoted the polarization of macrophages to M1-like phenotype and IL-6, TNF-α secretion as well as the pro-inflammatory differentiation of CD4+T cells into Th17 cells. Upon silencing the expression of TSP-1 in tMSCs, the pro-inflammatory effects of MEXsLPS were suppressed. Therefore, these findings uncovered TSP-1 as the principal factor in MEXsLPS pro-inflammatory regulation.

[Inhibitory effect and molecular mechanism of silencing S100A4 gene on the growth of transplanted tumor of human esophageal carcinoma EC-1 cells in nude mice].

OBJECTIVE: To study the effect of S100A4siRNA on tumor growth of xenografted human esophageal squamous cell carcinoma (ESCC) cell line EC1 in nude mice and explore its related molecular mechanism. METHODS: The xenografted tumor model was established in nude mice, and S100A4siRNA chemically synthesized was used to transfect the xenografted nude mice. The tumor growth was observed. The mRNA and protein expressions of S100A4, MMP-2 and E-cadherin in tumor after transfection with S100A4siRNA were detected by RT-PCR and immunohistochemistry. RESULTS: The tumor volume of S100A4siRNA transfection group was lower than that of nonsense siRNA transfection group and blank control group (P < 0.05), and tumor inhibitive ratio of S100A4siRNA group was higher than that of nonsense siRNA transfection group (P < 0.05). Furthermore, the mRNA and protein expressions of S100A4, MMP-2 in S100A4siRNA group were lower than those in nonsense siRNA group and control group (P < 0.05), the mRNA and protein expressions of E-cadherin in S100A4siRNA group were higher than those in nonsense siRNA group and control group (P < 0.05). CONCLUSION: S100A4siRNA could effectively lead to growth inhibition of xenografted human esophageal tumor in nude mice, with down regulation of MMP-2 and up regulation of E-cadherin, which provides theoretical basis for molecular therapy of ESCC.

Identification of key gene modules and genes in colorectal cancer by co-expression analysis weighted gene co-expression network analysis.

Colorectal cancer (CRC) has been one of the most common malignancies worldwide, which tends to get worse for the growth and aging of the population and westernized lifestyle. However, there is no effective treatment due to the complexity of its etiology. Hence, the pathogenic mechanisms remain to be clearly defined. In the present study, we adopted an advanced analytical method-Weighted Gene Co-expression Network Analysis (WGCNA) to identify the key gene modules and hub genes associated with CRC. In total, five gene co-expression modules were highly associated with CRC, of which, one gene module correlated with CRC significantly positive (R = 0.88). Functional enrichment analysis of genes in primary gene module found metabolic pathways, which might be a potentially important pathway involved in CRC. Further, we identified and verified some hub genes positively correlated with CRC by using Cytoscape software and UALCAN databases, including PAICS, ATR, AASDHPPT, DDX18, NUP107 and TOMM6. The present study discovered key gene modules and hub genes associated with CRC, which provide references to understand the pathogenesis of CRC and may be novel candidate target genes of CRC.

Klf5 acetylation regulates luminal differentiation of basal progenitors in prostate development and regeneration.

Prostate development depends on balanced cell proliferation and differentiation, and acetylated KLF5 is known to alter epithelial proliferation. It remains elusive whether post-translational modifications of transcription factors can differentially determine adult stem/progenitor cell fate. Here we report that, in human and mouse prostates, Klf5 is expressed in both basal and luminal cells, with basal cells preferentially expressing acetylated Klf5. Functionally, Klf5 is indispensable for maintaining basal progenitors, their luminal differentiation, and the proliferation of their basal and luminal progenies. Acetylated Klf5 is also essential for basal progenitors' maintenance and proper luminal differentiation, as deacetylation of Klf5 causes excess basal-to-luminal differentiation; attenuates androgen-mediated organoid organization; and retards postnatal prostate development. In basal progenitor-derived luminal cells, Klf5 deacetylation increases their proliferation and attenuates their survival and regeneration following castration and subsequent androgen restoration. Mechanistically, Klf5 deacetylation activates Notch signaling. Klf5 and its acetylation thus contribute to postnatal prostate development and regeneration by controlling basal progenitor cell fate.

[Effect of KISS-1 on invasive potential and proliferation of esophageal squamous carcinoma cell line EC-1].

OBJECTIVE: To investigate the effect of KISS-1 expression on the potential of invasion and proliferation of esophageal squamous carcinoma cell EC-1. METHODS: Protein and mRNA expressions of KISS-1 were evaluated by Western blot and RT-PCR in four esophageal carcinoma cell lines (EC-1, Eca109, EC9706 and TE-1). Using liposome-mediated transfection, an eukaryotic expression vector (pcDNA3.1-KISS-1) of KISS-1 gene was transfected into EC-1 cells. Boyden chamber model, MTT and clone formation assay were used to detect the potential of invasion and proliferation. RESULTS: Western blot and RT-PCR showed a baseline low level of expression of KISS-1 protein (0.715 +/- 0.109) and mRNA (0.670 +/- 0.176) in EC-1 cells. pcDNA3.1-KISS-1 expression vector was successfully transfected into EC-1 cells. Western blot and RT-PCR showed that the expression of KISS-1 protein (1.143 +/- 0.218) and mRNA (0.877 +/- 0.162) in EC-1 cells transfected with pcDNA3.1-KISS-1 were significantly higher than those transfected with the control vector pcDNA3.1 (0.745 +/- 0.130, 0.685 +/- 0.128; t = 3.850, 2.481, P < 0.05) and the control cells (0.855 +/- 0.184, 0.677 +/- 0.138; t = 2.275, 2.306, P < 0.05). Boyden chamber analysis showed that the invasiveness of the cells transfected with KISS-1 at 24 h (91.8 +/- 11.7), 48 h (117.8 +/- 11.1) and 72 h (139.2 +/- 11.8) were significantly reduced than that of the cells transfected with the control vector pcDNA3.1 (118.1 +/- 14.7, 141.7 +/- 13.2, 162.2 +/- 22.7; t = 3.153, 4.215, 3.569, P < 0.01) and the control cells (112.2 +/- 15.6, 138.1 +/- 13.0, 162.3 +/- 14.0; t = 4.154, 3.797, 2.702, P < 0.05). MTT showed that the proliferation potential of cells after transfection with KISS-1 at 48 h (0.517 +/- 0.127) and 72 h (0.394 +/- 0.137) were significantly reduced than that of cells transfected with the control vector pcDNA3.1 (0.636 +/- 0.186, 0.513 +/- 0.150; t = 2.054, 2.709, P < 0.05) and the control cells (0.646 +/- 0.135, 0.511 +/- 0.153; t = 2.276, 2.205, P < 0.05). Clone formation assay suggested that cells transfected with KISS-1 (157.2 +/- 36.4) showed significantly decreased clone formation than cells transfected with the control vector pcDNA3.1 (236.3 +/- 78.1; t = 3.441, P < 0.01) and the control cells (242.5 +/- 48.6; t = 2.250, P < 0.05). CONCLUSION: KISS-1 gene inhibits the potential of invasion and proliferation of EC-1 cells.

Retraction Note: RNAi targeting CXCR4 inhibits proliferation and invasion of esophageal carcinoma cells.

The Editor-in-Chief has retracted this article [1] because Fig. 3 shows overlap with Fig. 6 in [2], Fig. 2b in [3] and Fig. 6a in [4]. An investigation by Zhengzhou University has confirmed that these figures overlap. The data reported in this article are therefore unreliable.