Comparative detection of aberrantly methylated DNA in preoperative and postoperative stool from patients with colorectal cancers.

BACKGROUND: Early detection of colorectal cancer (CRC) is crucial to reducing tumor-related mortality. Evaluating aberrantly methylated DNA in stool is promising for CRC screening. However, DNA methylation in the colonic epithelium of background mucosa may compromise stool DNA (sDNA) test results. Thus, we compared aberrant methylation of cancer-related genes in preoperative and postoperative sDNA, with the aim of demonstrating that a cancer-specific methylated allele in sDNA originates from CRCs. METHODS: Patients who were to undergo CRC resection in Kyushu University Hospital during 2003-2010 were prospectively enrolled. Preoperative (pre) stool samples from 54 patients, postoperative (post) samples from 52 of the patients and tumor samples were collected. Aberrant promoter methylation of CDH4 and GATA5 was assessed in the primary tumors by methylation-specific polymerase chain reaction (MSP) and in stool samples by real-time MSP. REULTS: Aberrant methylation of CDH4 and/or GATA5 was detected in 45 of CRC tissue samples (83.3%) and identified in 23 pre sDNA samples (42.3%) from CRC patients. Aberrant methylation was not found in pre sDNA obtained from CRC patients without aberrant methylation of these genes or in post sDNA in any patient. The detection rate of methylated alleles did not correlate with depth of invasion or tumor stage. CONCLUSION: Our findings demonstrate that aberrantly methylated alleles identified in sDNA originate from CRCs. Although tumor-specific aberrant methylation is found in sDNA from patients harboring early and advanced CRC throughout the colon and rectum, the sensitivity of this test needs to be improved for early detection of CRC.

A CpG island methylator phenotype of colorectal cancer that is contiguous with conventional adenomas, but not serrated polyps.

A subset of colorectal cancers (CRCs) harbor the CpG island methylator phenotype (CIMP), with concurrent multiple promoter hypermethylation of tumor-related genes. A serrated pathway in which CIMP is developed from serrated polyps is proposed. The present study characterized CIMP and morphologically examined precursor lesions of CIMP. In total, 104 CRCs treated between January 1996 and December 2004 were examined. Aberrant promoter methylation of 15 cancer-related genes was analyzed. CIMP status was classified according to the number of methylated genes and was correlated with the clinicopathological features, including the concomitant polyps in and around the tumors. The frequency of aberrant methylation in each CRC showed a bimodal pattern, and the CRCs were classified as CIMP-high (CIMP-H), CIMP-low (CIMP-L) and CIMP-negative (CIMP-N). CIMP-H was associated with aberrant methylation of MLH1 (P=0.005) and with an improved recurrence-free survival (RFS) rate following curative resection compared with CIMP-L/N (five-year RFS rate, 93.8 vs. 67.1%; P=0.044), while CIMP-N tumors were associated with frequent distant metastases at diagnosis (P=0.023). No concomitant serrated lesions were present in the tumors, whereas conventional adenoma was contiguous with 11 (10.6%) of 104 CRCs, including four CIMP-H CRCs. CIMP-H was classified in CRCs by a novel CIMP marker panel and the presence of concomitant tumors revealed that certain CIMP-H CRCs may have arisen from conventional adenomas.

Esophageal ruptures: triage using the systemic inflammatory response syndrome score.

Esophageal rupture is a rare entity. Delay in the diagnosis and treatment may threaten the patient's life. The decision for surgical or nonsurgical treatment, however, remains controversial because advocates of both treatments have reported comparable results. To quantify the decision making, we suggest the systemic inflammatory response syndrome (SIRS) score for triage of an esophageal rupture. Using this criterion for 12 patients resulted in the survival of all of them. Therefore, we advocate use of the SIRS score for triage of an esophageal rupture.

[Chemotherapy for non-small cell lung cancer: split-dose administration of Cisplatin over four consecutive days and Vinorelbine ditartrate].

PURPOSE: At present, combination chemotherapy with Cisplatin (CDDP) and Vinorelbine ditartrate (VNR) is one of the standard regimens for non-small cell lung cancer (NSLC). To avoid renal damage by CDDP, hydration and diuretic are indicated. But elderly/postoperative patients who have reduced lung vessel capacity are a high-risk group for pulmonary edema/right heart failure by hydration. In our hospital, CDDP is administered on four consecutive days without large hydration. MATERIAL & METHODS: CDDP: 80 mg/m2 (over four consecutive days)without large hydration+VNR: 20 mg/m2 was administered 30 NSLC patients(Stage III A & IV). Serum concentration of CDDP was monitored. RESULT: Response rate was CR: 0 case; PR: 9 cases; SD: 16 cases; PD: 5 cases. Mean survival time (MST) was 292 days. The efficacy and prognosis are equivalent to a conventional CDDP+VNR regimen. On the other hand, side effects were reduced; neutrocytopenia (> Grade 3): 17%, renal dysfunction (>Grade 1): 17%. Mean serum concentrations of CDDP were accumulated day by day, 0.91 microg/mL(Day 1), 2.44 microg/mL(Day 4), but were all under the toxic threshold(8 microg/mL). CONCLUSION: Our regimen (CDDP given over four consecutive days without large hydration) may become a regimen for the high-risk patient.

Ferrocenylnaphthalene diimide-based electrochemical detection of methylated gene.

Ferrocenylnaphthalene diimide (FND)-based electrochemical hybridization assay was applied to the detection of methylated cytosine of DNA using the products obtained after treatment with bisulfite followed by polymerase chain reaction (PCR), where unmethylated cytosine is converted to thymine and methylated one to cytosine. Twenty-meric DNA probes for the methylated (cytosine) and unmethylated (thymine) types of the part of the promoter region of cyclin D-dependent protein kinase inhibitor, p16, gene (p16(Ink4a)) were used to be immobilized on the electrochemical array (ECA) chip. Using 1 microL of 10 ng/microL of methylated sample obtained from the methylation-specific PCR of methylated genome containing 10-times excess of unmethylated one, the methylated PCR sample could be detected by the identical electrochemical signals from the two DNA probes under the settled optimum hybridization conditions.

Familial pancreatic cancer: report of one Japanese family.

Most familial pancreatic carcinomas have been reported from European countries and the United States, and there has been only one report from Japan. A 50-year-old Japanese woman presented with a pancreatic head mass and underwent pylorus-preserving pancreatoduodenectomy with portal vein resection. The histological diagnosis was well-differentiated adenocarcinoma of the head of the pancreas. Her mother died of pancreatic head carcinoma, which had been shown on computed tomography at the age of 70 years. One of her uncles on her father's side had had pancreatic tail carcinoma, and at the age of 59, had undergone distal pancreatectomy, splenectomy, wedge resection of the liver, and partial resection of the colon. The histological diagnosis was moderately differentiated tubular adenocarcinoma of the pancreas. He had had a subtotal gastrectomy for early gastric cancer (tubular adenocarcinoma limited to the mucosa) at the age of 53. He died of recurrence of the pancreatic tail carcinoma 3 months after the distal pancreatectomy had been performed. This communication reports a second Japanese family with familial pancreatic cancer, as shown by pancreatic carcinomas in two first-degree relatives and in one third-degree relative.