RUNX3 promoter methylation in colorectal cancer: its relationship with microsatellite instability and its suitability as a novel serum tumor marker.

BACKGROUND/AIM: RUNX3 is a novel gastric cancer tumor suppressor. RUNX3 promoter hypermethylation is associated with many types of cancer, including colorectal cancer. Furthermore, the RUNX3 promotor is one of the CpG island methylator phenotype (CIMP)-specific promotors. CIMP is a distinct phenotype associated with microsatellite instability (MSI) in colorectal cancer. In this study, the suitability of the quantitative analysis of RUNX3 promoter hypermethylation as a novel serum tumor marker was investigated. Moreover, we investigated the relationship between RUNX3 promoter methylation and MSI in colorectal cancer. PATIENTS AND METHODS: A RUNX3 real-time quantitative methylation-specific PCR (RTQ-MSP) technique we developed was used to analyze the CpG sites in the RUNX3 promoter of 119 colorectal tumors and 344 sera from colorectal cancer patients. MSI analysis of 119 colorectal tumors was performed with five microsatellite markers (BAT25, BAT26, D5S346, D2S123, and D17S250). RESULTS: Proximal colon tumors exhibited significantly higher RUNX3 methylation than their paired normal tissues (p=0.0438). Analysis of the clinicopathological parameters revealed that a proximal location (p=0.0054), lymphatic invasion (p<0.0001), and an advanced pathological stage (p=0.0018) were associated with significantly higher RUNX3 methylation. Assessment of the relationship between RUNX3 methylation and tumor MSI revealed 11 out of 13 tumors with high-frequency MSI (85%) were positive for RUNX3 hypermethylation, significantly more than the tumors with low-frequency MSI or which were microsatellite stable (34%, p=0.0070). In preoperative sera from 344 colorectal cancer patients, significantly higher RUNX3 methylation was associated with lymphatic invasion (p=0.0487) and an advanced pathological stage (p=0.0466). Post-operative follow-up data revealed that recurrence cases exhibited significantly higher preoperative serum RUNX3 methylation than non-recurrence cases (p=0.0003). Concomitant analysis of carcinoembryonic antigen (CEA) levels in the preoperative sera showed that 17.7% (61/344) were CEA-negative but RUNX3 methylation-positive, which means assessing both serum RUNX3 methylation and CEA should improve diagnosis of colorectal carcinoma. CONCLUSION: RTQ-MSP-based quantification of serum RUNX3 methylation is useful for the detection and monitoring of colorectal cancer.

Quantitative analysis of tumor-derived methylated RUNX3 sequences in the serum of gastric cancer patients.

PURPOSE AND EXPERIMENTAL DESIGN: Using real-time quantitative methylation-specific PCR (RTQ-MSP), methylated RUNX3 sequences were quantified and the fractional concentrations of circulating tumor DNA in serum were determined, along with peripheral blood cells collected preoperatively, intraoperatively and postoperatively from 65 patients with gastric cancer. RESULTS: RTQ-MSP was sufficiently sensitive to detect RUNX3 methylation. Quantitative MSP data were expressed in terms of the methylation index, which was defined as the relative amount of methylated RUNX3 sequences divided by the concentration of methylated actin. High levels of methylated RUNX3 sequences were detected in the peripheral circulation of 29% (19 of 65) of the gastric cancer patients. The RUNX3 methylation index was concordant with cancer stage, histology, lymphatic and vascular invasion, and was more sensitive than carcinoembryonic antigen (CEA) as a biomarker. Twenty-nine percent (19 out of 65) of preoperative serum samples had methylated RUNX3 sequences, ranging from 5.2 to 1625955 (median quantity=43 m-index, sensitivity 95.5%, specificity 62.5%, AUC 0.8651). After surgical resection, the median RUNX3 methylation index in serum significantly decreased. These results demonstrate the clinical usefulness and effectiveness of peripheral blood RTQ-MSP for detecting and monitoring gastric cancer after treatment. Furthermore, 5 out of the 30 preoperative control samples of benign disease (cases of panperitonitis due to acute appendicitis or cholecystitis) showed transient RUNX3 methylation which decreased after the operation in accordance with recovery. CONCLUSION: Quantification of epigenetic changes in serum RUNX3 methylation using RTQ-MSP is useful for the detection and monitoring of gastric cancer.

Outcomes of total pelvic exenteration for colorectal cancer.

BACKGROUND/AIMS: Pelvic recurrence occurs in 4-33% of patients who have undergone a curative resection of primary rectal cancer and is thus a serious problem. However, the best treatment for primary rectal cancer remains unclear. In the present study was assessed the outcomes of total pelvic exenteration for colorectal cancer retrospectively. METHODOLOGY: In the present study was investigated the medical charts of 25 patients who underwent total pelvic exenteration for primary colorectal cancer (n = 12) or postoperative local recurrence of colorectal cancer (n = 13) at the Department of Surgery (Division of Digestive Surgery) of the Kyoto Prefectural University of Medicine between the years 1997-2005. RESULTS: The mean disease-free time interval between the first operation for primary colorectal cancer and total pelvic exenteration for the recurrence was 919 days (range, 203-3460 days). Total pelvic exenteration required a mean operation time of 940 minutes (range, 540-1395 minutes). The mean carcinoembryonic antigen (CEA) value was 25.5 ng/ml (range, 1-171.8 ng/ml). Five-year survival was achieved in 9 patients (36%) and inhospital death occurred in 3 patients (12%). The patients with curative resection survived significantly longer than the patients with non-curative resection. CONCLUSIONS: When curative resection is achieved, total pelvic exenteration for colorectal cancer can result in long-term survival.

[A case of huge advanced rectal cancer invaded into the surrounding organs resected successfully after preoperative chemotherapy with mFOLFOX6].

In the recent improvement in chemotherapy for advanced rectal cancer, a treatment for rectal cancer involving the surrounding organs has been well thought out. In this report, we described a case of advanced rectal cancer invaded into the surrounding organs was resected successfully after preoperative chemotherapy with mFOLFOX6. The case was a 74-year-old man with advanced rectal cancer (type 3). A close examination of the patient revealed a bowel movement disturbance. Bowel obstruction was treated with transverse colostomy. Then chemotherapy (mFOLFOX6) was performed six times. It was judged at first to be a huge tumor of 15 cm in diameter, which was unresectable due to invasion into the urinary bladder and sacrum. However, after mFOLFOX6 was enforced, the tumor was shrunk to about 5 cm in diameter (effect judgment PR). Then the tumor was successfully resected. A pathologic histology inspection of the tumor, judged to be Grade 2 prior to resection, revealed a differentiation type glandular carcinoma and a highly lymphocytic infiltration. These results suggested that an appropriate preoperative chemotherapy was useful for huge rectal cancers involving the surrounding organs such as urinary bladder and sacrum.

Huge ovarian metastasis from jejunal cancer occurring immediately after initial operation.

Correct preoperative diagnosis of jejunal cancer is difficult, and there are no clear guidelines for the postoperative follow-up to monitor the recurrence of jejunal cancer. This report describes asynchronous ovarian metastasis from jejunal cancer occurring after initial operation. The patient was a 68-year-old postmenopausal woman with a preoperatively correctly diagnosed jejunal cancer. Partial resection of the jejunum with regional lymph node dissection was performed, with both ovaries intact at the first operation. Three months later, vaginal bleeding and a large metastasis in the left ovary were detected, for which a left oophorectomy was performed. Peritoneal dissemination was also detected perioperatively, so that chemotherapy with 5-FU/leukovoline was started. The patient is still alive more than 12 months after the most recent operation. Our case demonstrates that it is necessary to pay careful attention to synchronous or non-synchronous metastases to the ovaries after operation for jejunal cancer in females. Consideration should also be given to the efficacy of oophorectomy for ovarian metastases and of intensive chemotherapy combined with jejunal resection for carcinoma because of the poor prognosis of these treatment modalities.

Massive lymphatic fluid leakage during laparoscopic lymphnode biopsy for abdominal malignancy.

This report concerns massive lymphatic fluid leakage during laparoscopic abdominal lymphnode biopsy for mesenteric tumor of non-Hodgkin's disease. Laparoscopic lymphnode biopsy was performed on a 58-year-old man who presented with a huge abdominal mass. The initial diagnosis was based on abdominal computed tomography, which revealed a large mass. This was followed by laparoscopic abdominal lymphnode biopsy for a definitive diagnosis. During the operation, massive lymphatic leakage up to about 300 mL occurred, but was stopped completely by electric coagulation. Histological examination indicated the mass to be a B-cell type non-Hodgkin's lymphoma. Hospitalization was uneventful, and the patient was discharged 7 days postoperatively to the Department of Internal Medicine for chemotherapy. This new endoscopic approach offers a useful alternative to the traditional transabdominal excision of intra-abdominal lymphnodes, although attention must be paid to possible complications including massive intraoperative lymphatic leakage.

Manifestation of bilateral huge ovarian metastases from colon cancer immediately after the initial operation: report of a case.

This report describes the case of a 34-year-old premenopausal woman in whom bilateral huge ovarian metastases were found immediately after initial surgery for sigmoid colon cancer. Both ovaries had been intact at the time of sigmoidectomy, but 2 months later, she complained of persistent vaginal bleeding, and large bilateral metastases were detected in both ovaries. Oophorectomy with intraperitoneal chemotherapy proved ineffective and the patient died 3 months later, after a second operation, from peritoneal dissemination. This case report serves to demonstrate the importance of searching for synchronous or nonsynchronous metastases to the ovaries after surgery for colon cancer in young women. Consideration should also be given to the feasibility of performing prophylactic oophorectomy or administering intensive chemotherapy in association with colon resections for carcinoma for premenopausal women because of the ineffectiveness of these modalities as treatment for metastatic disease.