Author Correction: Enah overexpression is correlated with poor survival and aggressive phenotype in gastric cancer.

In the version of this article originally submitted, Fig 5j included a mismatched image which had been accidentally inserted by the authors during collation of the data. The final data was derived from three independent experiments, and three repetitions for each experiment to ensure the repeatability, reliability and scientificity of the experiment. During this process, the fields overlapped to some extent, giving rise to the mistake. It does not alter any inferences drawn from the data, and the statistical graph is correct. This error has been corrected in both the PDF and HTML versions of the Article.

Enah overexpression is correlated with poor survival and aggressive phenotype in gastric cancer.

Enabled homolog (Enah), which is a member of the Ena/VASP family that also includes VASP (vasodilator-stimulated phosphoprotein) and Ena/VASP like, is a mammalian ortholog of Drosophila Enabled (Ena). An increasing number of studies demonstrated Enah overexpression is involved in human colorectal carcinomas, breast cancers and hepatocellular carcinoma. However, the significance of Enah expression in gastric cancer (GC) is poorly elucidated. Here, we demonstrate that Enah is upregulated in GC and associated with AJCC stage, depth of invasion and poor overall survival (OS). Knockdown of Enah inhibited GC cell proliferation and metastasis and vice versa. Further experiments suggested that p-Erk1/2, p-AKT, p-p65, Vimentin and Fibronectin were downregulated and E-cadherin was upregulated after Enah silencing, implicating altered functions in GC proliferation and metastasis. Thus, our study suggests that Enah is a harmful factor for GC and a novel target for GC treatment.

FOXK1 plays an oncogenic role in the development of esophageal cancer.

Forkhead box k1 (FOXK1) is a member of the FOX class of transcription factors and it is dysregulated in many solid tumors including hepatocellular carcinoma, gastric cancer, colorectal cancer and prostate cancer. However, the expression status of FOXK1 and its clinical significance in esophageal cancer (EC) is still uncertain. Our study aimed at investigating the significance of FOXK1 expression in human EC and its biological function in the development of EC. We found that FOXK1 was overexpressed in EC tissues compared with corresponding non-tumor tissues using immunohistochemistry. And high FOXK1 expression was related to poor differentiation of EC. The Kaplan-Meier curve indicated that high FOXK1 expression may result in poor prognosis of EC patients. Furthermore, overexpression of FOXK1 in EC9706 cell inhibited cell apoptosis and promoted cell proliferation and migration, and suppression of FOXK1 in EC109 cell obtained reverse results. Our data suggest that FOXK1 plays an oncogenic role in EC pathogenesis and can serve as a therapeutic target for patients with EC.

Increased expression of calponin 2 is a positive prognostic factor in pancreatic ductal adenocarcinoma.

Calponin 2 plays an important role in regulating actin cytoskeleton, which is critical for cell division and migration. Previous studies have demonstrated that calponin 2 inhibits prostate cancer cell proliferation and metastasis. However, the role of calponin 2 in pancreatic tumor growth, metastasis and patient survival remains unclear. Here, we demonstrate that the level of calponin 2 is a positive prognostic factor for patients with pancreatic ductal adenocarcinoma (PDAC). Patients with high calponin 2 expression in the tumor presented less lymph node metastasis and longer survival. Knockdown of calponin 2 facilitated pancreatic cancer cell proliferation and metastasis. Further experiments suggested that PI3K/AKT, NF-κB, Vimentin, Fibronectin, Snail and Slug were upregulated and E-cadherin was downregulated after calponin 2 was knocked down, implicating altered functions in PDAC proliferation and metastasis. In addition, we verified that calponin 2 functioned through inhibiting PI3K/AKT and NF-κB pathways. Our study suggests that the upregulation of calponin 2 in PDAC correlates to lower malignancy and presents a novel target for the development of new treatment.

Folate receptor alpha is associated with cervical carcinogenesis and regulates cervical cancer cells growth by activating ERK1/2/c-Fos/c-Jun.

Folate receptor alpha (FRα) is over-expressed in numerous epithelial malignancies, however, the association between FRα and cervical cancer remains unclear. The purpose of this study was to explore the effects of FRα on cervical cancer and its regulation of the ERK signaling pathway. In this case-control study, moderate/strong expression of FRα, phosphorylated ERK1/2 (p-ERK1/2), p-c-Fos, and p-c-Jun proteins was increased with the progressive severity of cervix lesions (P < 0.05). Moreover, the expression levels of p-ERK1/2, p-c-Fos, and p-c-Jun proteins was positively correlated with those of FRα protein in cervical squamous cell carcinoma (SCC) group (P < 0.05). In vitro experiment indicated down-regulation of FRα by siRNA suppressed cell proliferation, promoted cell apoptosis, induced cell cycle arrest at G0/G1 phase, and reduced expression of p-ERK1/2, p-c-Fos, and p-c-Jun proteins. The results suggest that FRα is associated with the progression of cervical cancer and regulates cervical cancer cells growth through phosphorylating ERK1/2, c-Fos, and c-Jun, which are key factors of the ERK signaling pathway. Therefore, FRα may be an effective target for early detection and therapy of cervical cancer.

Community physicians' knowledge of secondary prevention after ischemic stroke: a questionnaire survey in Shanxi Province, China.

BACKGROUND: This cross-sectional, questionnaire-based survey, conducted in Shanxi Province, China, evaluated the knowledge of community physicians of secondary prevention of ischemic stroke and transient ischemic attacks (TIAs). METHODS: A total of 1910 physicians practicing at 832 community-based clinics, hospitals and other care centers in 11 prefectures of Shanxi Province completed the questionnaires between 1 July and 30 September 2013. RESULTS: Over 90 % of participants were aware of the most common risk factors for stroke, but lifestyle-related factors were seen as of low or medium importance for secondary prevention. Only about 50 % of physicians were aware of the existence of commonly used stroke scales, and fewer said that they would use those scales in their clinical practice. There were slight differences in the responses to some of the questions on risk factors and stroke scales were associated with the physicians' gender, academic qualifications, practice duration and location. Less than half of the participants were aware of the secondary prevention recommendations included in the most recent guidelines. CONCLUSION: The survey revealed a huge gap in knowledge of current guidelines for secondary prevention of ischemic stroke and TIA among the physicians surveyed. Continuing education and training of community physicians, administered as a public health program, is needed to improve the healthcare of ischemic stroke and TIA patients.

ERK1/2 promoted proliferation and inhibited apoptosis of human cervical cancer cells and regulated the expression of c-Fos and c-Jun proteins.

Small-molecule inhibitors targeted MAPK have been wildly used for some cancer therapeutics as a biologically viable model, but no one has been used for cervical caner. ERK1/2, one of MAPK kinases, is expressed high in cervical cancer tissue. The aim of the present study was to evaluate the effects of ERK1/2 inhibitor U0126 on proliferation and apoptosis of cervical cancer cells and appraise the correlated mechanism of the effects. In this study, the cell proliferation of Hela and C33A cervical cancer cells was tested by Cell Counting Kit-8 (CCK8) assay and cell counting after treated with ERK1/2 inhibitor U0126. The cell cycle and apoptosis were evaluated by flow cytometry (FCM). The protein levels of ERK1/2 and c-Fos and c-Jun were detected by Western blot. The results indicated that after down-regulating ERK1/2 proteins with the inhibitor U0126, Hela and C33A cells proliferation was inhibited, cell apoptosis was promoted, the proportions of G0/G1 stage in cell cycle increased, and G2/M stages decreased. After down-regulating ERK1/2 proteins of Hela and C33A cells, the expression levels of p-c-Fos protein decreased, while p-c-Jun protein increased. The results of this study indicated that ERK1/2 may promote the development of cervical cancer cells, suggesting ERK1/2 inhibitor may be used as an effective target for cervical cancer therapies working for. It might inhibit cervical cancer cells growth via regulating the transcription factors expression of c-Fos and c-Jun.

Folate deficiency and FHIT hypermethylation and HPV 16 infection promote cervical cancerization.

Fragile histidine triad (FHIT) is a suppressor gene related to cervical cancer through CpG island hypermethylation. Folate is a water-soluble B-vitamin and an important cofactor in one-carbon metabolism. It may play an essential role in cervical lesions through effects on DNA methylation. The purpose of this study was to observe effects of folate and FHIT methylation and HPV 16 on cervical cancer progression. In this study, DNA methylation of FHIT, serum folate level and HPV16 status were measured using methylation-specific polymerase chain reaction (MSP), radioimmunoassay (RIA) and polymerase chain reaction (PCR), respectively, in 310 women with a diagnosis of normal cervix (NC, n=109), cervical intraepithelial neoplasia (CIN, n=101) and squamous cell carcinoma of the cervix (SCC, n=101). There were significant differences in HPV16 status (χ2=36.64, P<0.001), CpG island methylation of FHIT (χ2=71.31, P<0.001) and serum folate level (F=4.57, P=0.011) across the cervical histologic groups. Interaction analysis showed that the ORs only with FHIT methylation (OR=11.47) or only with HPV 16 positive (OR=4.63) or with serum folate level lower than 3.19ng/ml (OR=1.68) in SCC group were all higher than the control status of HPV 16 negative and FHIT unmethylation and serum folate level more than 3.19ng/ml (OR=1). The ORs only with HPV 16 positive (OR=2.58) or with serum folate level lower than 3.19ng/ ml (OR=1.28) in CIN group were all higher than the control status, but the OR only with FHIT methylation (OR=0.53) in CIN group was lower than the control status. HPV 16 positivity was associated with a 7.60-fold increased risk of SCC with folate deficiency and with a 1.84-fold increased risk of CIN. The patients with FHIT methylation and folate deficiency or with FHIT methylation and HPV 16 positive were SCC or CIN, and the patients with HPV 16 positive and FHIT methylation and folate deficiency were all SCC. In conclusion, HPV 16 infection, FHIT methylation and folate deficiency might promote cervical cancer progression. This suggests that FHIT may be an effective target for prevention and treatment of cervical cancer.

Down-regulation of FRα inhibits proliferation and promotes apoptosis of cervical cancer cells in vitro.

Folate receptor alpha (FRα) mediates folate uptake by endocytosis, and while folate is essential to DNA methylation and synthesis and may have an important role in proliferating cells. FRα is known to be expressed in rapidly proliferating cells, including many cancer cell lines, but there has been no systematic assessment of expression in cervical cancer cell lines. The aim of the present study was to evaluate the effects of FRα on proliferation and apoptosis of cervical cells and correlation mechanism. In this study, we investigated the biological function of FRα in Hela cells using RNA interference. Cell proliferation was evaluated by Cell Counting Kit-8 (CCK8) assay, while cell cycling and apoptosis were assessed by flow cytometry, mRNA levels by real time- PCR and protein levels of FRα, c-Fos and c-Jun by Western blotting. The results revealed that FRα was highly expressed in Hela cells and its silencing with a small interfering RNA (siRNA) inhibited cell proliferation and induced cell apoptosis, arresting the cell cycle in G0/G1 stages while decreasing the proportion in S and G2/M stages, and suppressed the expression levels of c-Fos and c-Jun. In conclusion, the results of this study indicated that FRα down-regulation might be capable of suppressing cervical cancer cell proliferation and promoting apoptosis. It suggested that FRα might be a novel therapeutic target for cervical cancer.

[Influence of folate on fragile histidine triad gene expression, cell proliferation and apoptosis in cervical cancer cell lines].

OBJECTIVE: To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation, expression of mRNA and protein of fragile histidine triad (FHIT)gene in cervical cancer cells. METHODS: Cervical cancer cell lines including CaSki (HPV16-positive) and C33A (HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR (MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR, respectively. RESULTS: Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.98, P < 0.001) and the apoptosis rate increased (C33A:r = 0.98, P < 0.001; CaSki:r = 0.99, P < 0.001) along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1 µg/ml and 10 µg/ml, partially positive at 100 µg/ml and 250 µg/ml, but negative at 500 µg/ml and 1 000 µg/ml in both C33A and CaSki cells. Comparing with the control group, the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells, and showing that the FHIT gene mRNA expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.94, P < 0.001) and protein expression (C33A:r = 0.96, P < 0.001; CaSki:r = 0.97, P < 0.001) both increased along with the increase of folate concentration. CONCLUSION: Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro, so would reverse the aberration mRNA and protein expression of FHIT gene.

[Effect of DNA methyltransferase 1 and methyl-CpG-binding protein 2 abnormal expression on cervical lesions and related interaction].

OBJECTIVE: To explore the effect of DNA methyltransferase 1 (DNMT1) and methyl-CpG-binding protein 2 (MeCP2) on cervical cancer and cervix precancerous lesion. METHODS: 74 patients with cervix squamous cell carcinoma(SCC), 52 patients with cervical intraepithelial neoplasm I (CIN I), 60 patients with cervical intraepithelial neoplasm II - II (CIN II-III)and 58 patients with histologically diagnosed cervix inflammation(CI), were included in this study. Information as demography, reproductive history, life style, HPV infection were collected. Western Blot were used to detect the expression of DNMT1 protein and MeCP2 protein. Real-time PCR was used to detect the expression of DNMT1 and MeCP2 mRNA. RESULTS: Levels of DNMT1 and MeCP2 protein expression increased gradually with the deterioration of cervical lesion (H = 94.33, P < 0.001;F = 21.580, P < 0.001). Along with the deterioration of cervical lesion, levels of DNMT1 and MeCP2 mRNA expression were gradually increasing( F = 4.758, P = 0.003; F = 7.804, P < 0.001). Data from Correlation analysis showed that both protein (r = 0.287, P < 0.001) and mRNA(r = 0.179, P = 0.005)were positive correlated with DNMT1 and MeCP2. RESULTS: of our study indicated that there was an additive interaction between high-expression of DNMT1 protein and high-expression of MeCP2 protein in SCC or CIN II-III. However, there was an additive interaction between high-expression of DNMT1 mRNA and high-expression of MeCP2 mRNA in SCC or CIN II-III. CONCLUSION: Results from our study revealed the fact that both high expression of DNMT1 protein and high expression of MeCP2 protein could increase the risk of cervix cancerization. According to our findings, there might be a synergistic action existed between DNMT1 and MeCP2 during the progression of cervix cancelation.