Testing for DNA methylation has potential in cancer screening. Most previous studies of DNA methylation in cervical cancer used a candidate gene approach. The aim our study was to identify novel genes that are methylated in cervical cancers and to test their potential in clinical applications. We did a differential methylation hybridization using a CpG island (CGI) microarray containing 8640 CGI tags to uncover methylated genes in squamous cell carcinomas (SCC) of the uterine cervix. Pooled DNA from cancer tissues and normal cervical swabs were used for comparison. Methylation-specific polymerase chain reaction, bisulfite sequencing and reverse transcription polymerase chain reaction were used to confirm the methylation status in cell lines, normal cervices (n 5 45), low-grade lesions (n 5 45), high-grade lesions (HSIL; n 5 58) and invasive squamous cell carcinomas (SCC; n 5 22 from swabs and n 5 109 from tissues). Human papillomavirus (HPV) was detected using reverse line blots. We reported 6 genes (SOX1, PAX1, LMX1A, NKX6-1, WT1 and ONECUT1) more frequently methylated in SCC tissues (81.5, 94.4, 89.9, 80.4, 77.8 and 20.4%, respectively) than in their normal controls (2.2, 0, 6.7, 11.9, 11.1 and 0%, respectively; p < 0.0001). Parallel testing of HPV and PAX1 methylation in cervical swabs confers an improved sensitivity than HPV testing alone (80% vs. 66%) without compromising specificity (63% vs. 64%) for HSIL/SCC. Testing PAX1 methylation marker alone, the specificity for HSIL/SCC is 99%. The analysis of these novel DNA methylations may be a promising approach for the screening of cervical cancers. ' 2008 Wiley-Liss, Inc.Key words: cervical cancer; epigenetics; hpv; methylation; microarray In addition to genetic changes, epigenetic alterations such as DNA methylation and histone modifications can result in heritable gene silencing without changes to genetic sequences and are recognized as important causes of cancer.1-3 DNA methylation mostly occurs at the 5 0 cytosine in the palindromic sequence, 5 0 -CpG-3 0 . CpG islands are CpG-rich areas of 1 kb that are usually located in the vicinity of genes, often near the promoters of widely expressed genes.4,5 Methylation of CpG sites in the human genome is catalyzed by a family of DNA methyltransferases (DNMTs). DNMT1 is a maintenance methyltransferase with a preference for hemimethylated DNA whereas DNMT3a and DNMT3b are de novo methyltransferases with approximately equal preferences for methylated and unmethylated DNA.2,6 The addition of methyl groups by DNMTs recruits complexes with transcription repressors that modify histones and thus silence genes. Global DNA hypomethylation and site-specific hypermethylation result in genomic instability and transcriptional gene inactivation, respectively, both of which are associated with cancer. 7,8 As epigenetic silencing of tumor suppressor genes by promoter hypermethylation is commonly observed in human cancers, DNA methylation could serve as a marker for early diagnosis of cancer and as a means of assessing the prognos...

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Aberrant CpG island hypermethylation is a common finding of cancers, which might be detectable in the tissue or serum of affected patients. We analyzed DNA methylation by methylationspecific polymerase chain reaction of 7 genes, which included secreted frizzled receptor proteins 1, 2, 4, 5 (SFRP1, 2, 4, 5), SRYbox 1 (SOX1), paired box gene 1 (PAX1) and LIM homeobox transcription factor 1, alpha (LMX1A) in primary tumor samples from 126 patients with ovarian cancer, 75 with a benign tumor and 14 with borderline malignancy of an ovarian tumor, and in the serum from 26 patients with ovarian cancer and 20 with a benign tumor. Six of 7 genes had higher methylation rates in patients with ovarian cancer than in borderline malignancy or benign tumor (p < 0.001). The methylation of SFRP1, SFRP2, SOX1 and LMX1A genes correlated with recurrence and overall survival of ovarian cancer patients. Combining the data for SFRP1, SFRP2 and SOX1 genes gave a relative risk for recurrence of 3.19 (p 5 0.013) in patients with at least one gene methylation, and combining the data for SFRP1, SOX1 and LMX1A gave an RR for cancer-related death of 6.09 (p 5 0.010). Methylation analysis of tissues and serum revealed a significant correlation (kappa values, 0.332-0.598) and a highly sensitivity and specificity rates (73.08 and 75%) as a screening marker. In conclusion, promoter hypermethylation of specific genes in critical pathways is common in ovarian cancer and has potential as a prognostic factor and a promising serum marker for early screening. ' 2008 Wiley-Liss, Inc.Key words: epigenetics; CpG islands; ovarian cancer; methylationspecific PCR About 190,000 new cases and 114,000 deaths from ovarian cancer are estimated to occur annually in the world. Ovarian cancer is the second most common gynecological malignancy and the fifth leading cause of cancer-related deaths among women in the United States. 1 Because there are no obvious symptoms in the early stage, 70-75% of patients are diagnosed in the advanced stages, and these patients have a low (<20%) 5-year survival rate. 2,3 Elevated serum concentration of cancer antigen 125 (CA125) is often associated with epithelial ovarian cancer but can also be associated with multiple benign disorders. 4 To be effective for screening, a marker requires high sensitivity for early detection and sufficient specificity to protect patients with false-positive results from unwarranted diagnostic evaluations. The development of useful molecular markers for ovarian cancer screening or prognostic prediction is warranted because of the low survival rate in patients diagnosed in the late stage of the disease.Candidate molecular markers include circulating DNA, RNA, or proteins produced from genetic or epigenetic modifications. As in other human cancers, loss of function of tumor suppressor genes (TSGs) caused by gene mutation, deletion or loss of heterozygosity are well-known genetic changes that induce tumor initiation and progression. However, these do not seem to be the major mechanism of TSG inactivation...

Highlights d SARS-CoV-2 genome sequencing and phylogenetic analyses identify 35 recurrent mutations d Association with 117 clinical phenotypes reveals potentially important mutations d D500-532 in Nsp1 coding region correlates with lower viral load and serum IFN-b d Viral isolates with D500-532 mutation induce lower IFN-I response in the infected cells

Oncogenic activation of the Wnt signaling pathway is common in cancers, but mutation of b-catenin in ovarian cancer is rare. In addition to genetic events, epigenetic modification of secreted frizzled-related protein (SFRP) family has been shown to be important in regulating Wnt signaling. Although high degree of homology is observed in the same family, different SFRPs may have opposing effects on the same process. We reported recently that a Wnt antagonist, SFRP5, is downregulated frequently through promoter hypermethylation and that this hypermethylation is associated with overall survival in ovarian cancer. The aim of this study was to analyze the function of SFRP5 in ovarian cancer. Functional assays including measuring cell proliferation, invasion, colony formation and xenograft were performed using ovarian cancer cell lines with overexpression of SFRP5 or a short hairpin RNA silencing. The methylation status of SFRP5 in relation to cisplatin resistance in ovarian cancer patients was analyzed. Restoration of the expression of SFRP5 attenuated Wnt signaling in ovarian cancer cells and suppressed cancer cell growth, invasion of cells and tumorigenicity in mice. These effects were independent of the canonical pathway. The expression of SFRP5 inhibited epithelial-mesenchymal transition (EMT). The restoration of SFRP5 downregulated AKT2 and sensitized ovarian cancer cells to chemotherapy. These effects are consistent with the poor response to platinumbased chemotherapy in patients with methylation of SFRP5. Our data suggested that epigenetic silencing of SFRP5 leads to oncogenic activation of the Wnt pathway and contributes to ovarian cancer progression and chemoresistance through the TWIST-mediated EMT and AKT2 signaling.Ovarian cancer is the second most common gynecological malignancy and the fifth leading cause of cancer-related death among women in the United States; nearly 60% of women with ovarian cancer eventually succumb to the disease.

A recent hypothesis for cancer chemoresistance posits that cytotoxic survival of a subpopulation of tumor progenitors drives the propagation of recurrent disease, underscoring the need for new therapeutics that target such primitive cells. To discover such novel compounds active against drug-resistant ovarian cancer, we identified a subset of chemoresistant ovarian tumor cells fulfilling current definitions of cancer-initiating cells from cell lines and patient tumors using multiple stemness phenotypes, including the expression of stem cell markers, membrane dye efflux, sphere formation, potent tumorigenicity, and serial tumor propagation. We then subjected such stem-like ovarian tumor-initiating cells (OTIC) to high-throughput drug screening using more than 1,200 clinically approved drugs. Of 61 potential compounds preliminarily identified, more stringent assessments showed that the antihelmintic niclosamide selectively targets OTICs in vitro and in vivo. Gene expression arrays following OTIC treatment revealed niclosamide to disrupt multiple metabolic pathways affecting biogenetics, biogenesis, and redox regulation. These studies support niclosamide as a promising therapy for ovarian cancer and warrant further preclinical and clinical evaluation of this safe, clinically proven drug for the management of this devastating gynecologic malignancy.

ObjectivesAn increased risk of tuberculosis (TB) has been reported in patients treated with TNF-α antagonists, an issue that has been highlighted in a WHO black box warning. This review aimed to assess the risk of TB in patients undergoing TNF-α antagonists treatment.MethodsA systematic literature search for randomised controlled trials (RCTs) was performed in MEDLINE, Embase and Cochrane library and studies selected for inclusion according to predefined criteria. ORs with 95% CIs were calculated using the random-effect model. Subgroup analyses considered the effects of drug type, disease and TB endemicity. The quality of evidence was assessed using the Grades of Recommendation, Assessment, Development and Evaluation (GRADE) approach.Results29 RCTs involving 11 879 patients were included (14 for infliximab, 9 for adalimumab, 2 for golimumab, 1 for etanercept and 3 for certolizumab pegol). Of 7912 patients allocated to TNF-α antagonists, 45 (0.57%) developed TB, while only 3 cases occurred in 3967 patients allocated to control groups, resulting in an OR of 1.94 (95% CI 1.10 to 3.44, p=0.02). Subgroup analyses indicated that patients of rheumatoid arthritis (RA) had a higher increased risk of TB when treated with TNF-α antagonists (OR 2.29 (1.09 to 4.78), p=0.03). The level of the evidence was recommended as ‘low’ by the GRADE system.ConclusionsFindings from our meta-analysis indicate that the risk of TB may be significantly increased in patients treated with TNF-α antagonists. However, further studies are needed to reveal the biological mechanism of the increased TB risk caused by TNF-α antagonists treatment.

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