[A case report of endosonography used for the diagnosis of early gastric cancer and gastrointestinal stromal tumor].

A 79-year-old man was referred to our hospital complaining of weight loss. Esophagogastroduodenoscopy revealed a flat, elevated lesion of 30-mm diameter located on the posterior wall of the upper gastric body. The lesion was histologically diagnosed as a well-differentiated adenocarcinoma. Endosonography showed a submucosal tumor under the carcinoma, which was highly suspicious of a gastrointestinal tumor derived from the muscle layer. Adenocarcinoma was diagnosed as an intramucosal lesion, but laparoscopic fundectomy was performed because of the submucosal tumor. Pathological diagnosis was U, less, pType 0-IIa, 18×30 mm, pT1b (SM1), tub2, int, INFa, ly0, v0, pN0, H0, P0, CYX, M0, pStage IA, gastrointestinal stromal tumors, prognostic group 1.

Methylation status of normal background mucosa is correlated with occurrence and development of neoplasia in the distal colon.

The aim of this study is to evaluate the methylation status of normal colonic mucosa in relation to the stage of neoplasia arising from the mucosa. The methylation status of 2 age-related loci (ESR1 and MYOD1) and global methylation (the mean of Alu and Sat2) in the normal colonic mucosa of 156 patients with and without colorectal neoplasia were examined. The distal colon and proximal colon were analyzed separately because neoplasia is biologically and clinically different between these sites. The methylation status was determined by MethyLight using percentage of methylated reference (PMR). In the distal colon, methylation of the age-related loci decreased as the stage of neoplasia increased (patients with no neoplasia or with adenoma < or =9 mm versus patients with advanced adenoma or with invasive cancer: ESR1-PMR median, 21.0 versus 15.7; P = .015; MYOD1-PMR median, 5.35 versus 3.80; P = .0037, respectively). Interestingly, global methylation was inversely correlated with the stage of neoplasia (59.7 versus 61.5; P = .054). In contrast, the proximal colon showed no significant correlations. The methylation of MYOD1 in the normal mucosa was significantly correlated with K-ras mutation in neoplastic tissue arising from the mucosa. Specific epigenetic changes in normal colonic mucosa may be correlated with the occurrence and development of neoplasia in the distal colon.

CO(2) insufflation for potentially difficult colonoscopies: efficacy when used by less experienced colonoscopists.

AIM: To clarify the effectiveness of CO(2) insufflation in potentially difficult colonoscopy cases, particularly in relation to the experience level of colonoscopists. METHODS: One hundred twenty potentially difficult cases were included in this study, which involved females with a low body mass index and patients with earlier abdominal and/or pelvic open surgery or previously diagnosed left-side colon diverticulosis. Patients receiving colonoscopy examinations without sedation using a pediatric variable-stiffness colonoscope were divided into two groups based on either CO(2) or standard air insufflation. Both insufflation procedures were also evaluated according to the experience level of the respective colonoscopists who were divided into an experienced colonoscopist (EC) group and a less experienced colonoscopist (LEC) group. Study measurements included a 100-mm visual analogue scale (VAS) for patient pain during and after colonoscopy examinations, in addition to insertion to the cecum and withdrawal times. RESULTS: Examination times did not differ, however, VAS scores in the CO(2) group were significantly better than in the air group (P < 0.001, two-way ANOVA) from immediately after the procedure and up to 2 h later. There were no significant differences between either insufflation method in the EC group (P = 0.29), however, VAS scores for CO(2) insufflation were significantly better than air insufflation in the LEC group (P = 0.023) immediately after colonoscopies and up to 4 h afterwards. CONCLUSION: CO(2) insufflation reduced patient pain after colonoscopy in potentially difficult cases when performed by LECs.

Methylation of estrogen receptor 1 in colorectal adenomas is not age-dependent, but is correlated with K-ras mutation.

The promoter region of estrogen receptor 1 (ESR1) has been shown to be methylated in normal colorectal mucosa in an age-dependent manner. However, the methylation of this region in colorectal tumors has not sufficiently been investigated. The methylation status of ESR1 in 105 colorectal adenoma tissues was examined by MethyLight and presented as the percentage of methylated references (PMR). Factors that affect the PMR of ESR1 in adenomas were determined using parameters including patient age, sex, past history of malignancy, family history of colorectal cancer, smoking and drinking habits, clinical characteristics of adenomas (location, size, macroscopic appearance, and histology), and K-ras mutation. Multiple linear regression revealed that the PMR was not correlated with patient age. K-ras mutation was significantly correlated with the higher methylation status of ESR1 in adenoma (t-value = 3.21, P = 0.0018), whereas alcohol exposure was significantly correlated with lower methylation status (t-value = -2.37, P = 0.02). Because methylation of O6-methylguanine DNA methyltransferase (MGMT) has been reported to be correlated with K-ras G-to-A transition, methylation of ESR1 was compared with that of MGMT with regard to K-ras mutation. Contrary to expectations, methylation of MGMT was not significantly correlated with K-ras G-to-A transition, but that of ESR1 was strongly correlated with K-ras G-to-A transition. Thus, the methylation status of ESR1 in adenomas was not correlated with patient age, but was associated with K-ras mutation, suggesting that methylation of ESR1 in tumors functions differently from that in normal colon mucosa.

Age-related methylation in normal colon mucosa differs between the proximal and distal colon in patients who underwent colonoscopy.

OBJECTIVES: To examine the difference in the methylation status in normal colon mucosa between the proximal and distal colon, in relation to the correlation between the methylation status of normal mucosa and characteristics of neoplasia. DESIGN AND METHODS: Paired biopsy specimens of normal mucosa from the proximal and distal colon of 82 patients who underwent colonoscopy were obtained. The methylation status of the promoter region of estrogen receptor 1 (ESR1) and myogenic differentiation 1 (MYOD1) was examined. RESULTS: Normal mucosa was more highly methylated in the distal than in the proximal colon in both ESR1 and MYOD1 loci (p<0.0001 and p=0.0009, respectively). Advanced characteristics of polyps in the distal colon were frequently observed in patients with lower methylation of ESR1 in the distal colon normal mucosa. CONCLUSIONS: Methylation levels in normal mucosa differ between the proximal and distal colon, and lower methylation of ESR1 in the distal colon normal mucosa may correlate with advanced features of neoplasia in the distal colon.

Decreased expression of hMLH1 correlates with reduced 5-fluorouracil-mediated apoptosis in colon cancer cells.

Patients with sporadic microsatellite instable colorectal cancers, in most of which the function of the hMLH1 mismatch repair gene is impaired, do not gain a survival benefit from 5-fluorouracil (5-FU)-based chemotherapy. However, the effect of hMLH1 on the cytotoxicity induced by 5-FU has not yet been sufficiently confirmed. In this study, we assessed the effect of hMLH1 on cytotoxicity and apoptosis induced by 5-FU using newly developed cell lines. We constructed two cell lines: SW480 (originally hMLH1-proficient), in which the expression of hMLH1 was reduced using a small interfering RNA (siRNA) technique, and HCT116 (originally hMLH1-deficient), in which the expression of hMLH1 can be regulated by doxycycline. Using these cell lines, a clonogenic survival assay, 4',6-diamidino-2-phenylindole (DAPI) staining and an Annexin-V assay were performed. Moreover, the incorporation of 5-FU into DNA was determined using tritium-labeled 5-FU. In both of our two cell lines, hMLH1-deficient cells exhibited approximately 2.4-fold clonal surviving fraction compared to hMLH1-proficient cells for 10 days after the administration of 5-FU. Additionally, hMLH1-deficient cells treated with 5-FU exhibited 34-45% less apoptosis than hMLH1-proficient cells according to the results of DAPI staining and Annexin-V assay. Furthermore, hMLH1-deficient cells treated with 5-FU exhibited an approximately 2-fold greater incorporation of 5-FU into DNA than control cells, suggesting that the recognition of 5-FU-incorporated DNA is impaired in hMLH1-deficient cells, resulting in reduced apoptosis. Our conclusions were that decreased expression of hMLH1 in colon cancer cells reduced the apoptosis induced by 5-FU, suggesting that hMLH1 is a key determinant of 5-FU chemosensitivity.

Laterally spreading type of colorectal adenoma exhibits a unique methylation phenotype and K-ras mutations.

BACKGROUND & AIMS: Laterally spreading tumors (LST), characterized by superficial extension along the colonic lumen, have recently been detected by colonoscopy. However, genetic and epigenetic characteristics of these tumors were scarcely reported. METHODS: A total of 205 sporadic colorectal adenoma tissues (157 protruded-type, 23 granular-type LST (G-LST), 12 flat-type LST (F-LST), and 13 flat-type smaller than 1 cm) were collected. CpG island methylator phenotype (CIMP) was determined by examination of methylation status at p16, methylated in tumor (MINT) 1, 2, 12, and 31 loci. K-ras codon 12 and 13 point mutations were also examined. The relationship between macroscopic appearance and CIMP status or K-ras mutations was analyzed. RESULTS: Among adenomas larger than 1 cm, CpG island methylation involving 2 or more loci (CIMP-high) was more likely to be observed in G-LST (14/23, 61%) than in protruded-type adenomas (18/73, 25%) (P = .002). The prevalence of K-ras mutations in G-LST (18/23, 78%) was significantly higher than that in protruded-type adenomas (18/73, 25%) (P < .0001). Moreover, the prevalence of CIMP-high and K-ras mutations in G-LST located in the proximal colon was much higher (11/13, 85%; and 12/13, 92%, respectively). In contrast, F-LST exhibited low prevalence of CIMP-high (1/12, 8%) and K-ras mutations (2/12, 16%). CONCLUSIONS: High prevalence of CIMP-high and K-ras mutations in G-LST, especially in the proximal colon, could strongly suggest that G-LST appearance is associated with a unique carcinogenic pathway.