Laparoscopy-assisted total gastrectomy for a case of MALT lymphoma: a case report.

A case of laparoscopy-assisted total gastrectomy with regional lymph node dissection for MALT (mucosa-associated lymphoid tissue) lymphoma is reported. The patient was a 45-year-old male with MALT lymphoma. Eradication of Helicobacter pylori as the initial therapy had failed to halt progression of the tumor. Considering certainty of the second therapy, he was transferred to surgical treatment. Total gastrectomy was required and this was performed by the laparoscopy-assisted approach. The postoperative course was satisfactory and the patient has been disease-free for 5 years after the operation. Laparoscopic operation could be a useful option in the second-line treatment for MALT lymphoma after failure to eradicate H. pylori.

Molecular detection of circulating esophageal squamous cell cancer cells in the peripheral blood.

PURPOSE AND EXPERIMENTAL DESIGN: To detect surgically resectable tumors earlier and improve the prognosis of esophageal squamous cell carcinoma patients, we examined deltaNp63 expression that was specific for squamous cell carcinoma in blood samples obtained from 43 esophageal cancer patients. RESULTS: Seventeen of 33 (52%) patients with primary esophageal squamous cell carcinoma and 6 of 10 (60%) patients with postoperative recurrent squamous cell carcinoma had detectable deltaNp63 expression in their peripheral blood using deltaNp63-specific reverse transcription-PCR. Furthermore, deltaNp63 is a more sensitive marker compared with other commonly used tumor markers such as squamous cell carcinoma-associated antigen and carcinoembryonic antigen. CONCLUSIONS: This approach would be potentially useful for the monitoring of patients with this aggressive disease.

A cancer-prone case with a background of methylation of p16 tumor suppressor gene.

PURPOSE AND EXPERIMENTAL DESIGN: To date, the presence of p16 gene promoter methylation associated with loss of protein expression has been demonstrated frequently in digestive tract cancers. In this study, we tested for the methylation status of p16 promoter in normal tissue specimens using the methylation-specific PCR technique to examine whether p16 methylation already existed in the background of tumors. RESULTS: Aberrant promoter methylation of p16 gene was detected in 1 of 40 esophageal and 1 of 69 gastric and no colorectal epithelium specimens, and these 2 specimens were derived from the same patient. We also found the same methylation change in both tumor and blood cell DNA. CONCLUSION: These results suggested that the p16 gene was inactivated by methylation in normal background cells of this patient and that other additional factors may promote tumor development in his esophageal and gastric tissues.

PGP9.5 as a marker for invasive colorectal cancer.

PURPOSE AND EXPERIMENTAL DESIGN: We proved recently that PGP9.5-negative pancreatic cancer patients had significantly better survival rates compared with those who were PGP9.5 positive, and PGP9.5 may be a novel marker for indicating the prognosis of pancreatic cancer patients. In this study, we examined the expression of PGP9.5 in primary colorectal cancers using immunohistochemistry and correlated the result with the clinicopathological features. RESULTS: Of 74 colorectal cancer specimens examined, 33 cases (46%) showed positive staining with PGP9.5 in most tumor cells, whereas no PGP9.5 expression was detected in adjacent normal epithelium. Subsequently, we correlated PGP9.5 expression in tumors with the clinicopathological features of affected patients and found two significant differences in maximal tumor size and the extent of tumor (P = 0.035 and 0.019, respectively). CONCLUSION: This result suggests that PGP9.5 expression is related to tumor progression and may be useful as a marker for invasive colorectal cancer.

Telomerase activity as prognostic factor in gastrointestinal stromal tumors of the stomach.

BACKGROUND/AIMS: Although size and mitotic counts have been reliable predictors of clinical outcome, identification of gastrointestinal stromal tumor (GIST) of the stomach with a metastatic potential through hematoxylin and eosin staining is not always accurate. Telomerase activity, often detected in malignant tumors, may have a role as a marker for high-grade malignancy. METHODOLOGY: Immunostaining with antibodies against KIT protein, CD34, and other molecules that are relevant for evaluation of cell differentiation and proliferation was performed for 36 primary gastric submucosal tumors to confirm the diagnosis of GIST. DNA was extracted from the surgically resected specimens of 24 of 36 patients, and c-kit mutation was analyzed by direct sequencing after PCR amplification of the exon 11. Telomerase activity was quantitatively evaluated for all 36 patients using fluorescence-based telomeric amplification assay protocol (TRAP) analysis. RESULTS: c-kit mutation was observed in 58% of the patients evaluated. Telomerase activity was detected in 14 specimens (39%), but not in the specimens without c-kit mutation. All 5 patients who suffered from metastatic or recurrent disease exhibited c-kit mutation and a high level of telomerase activity. CONCLUSIONS: Measurement of telomerase activity, along with c-kit mutation analysis, is useful for identifying GIST with a potential for malignant behavior.

Molecular detection of p16 promoter methylation in the serum of colorectal cancer patients.

Assays based on the molecular detection of genetic changes in serum have been shown as potential diagnostic tools for colorectal cancer. We examined the methylation status of p16 in colorectal cancers using methylation-specific PCR (MSP). Forty-four of 94 (47%) cancer DNA exhibited abnormal promoter methylation of p16 gene while no corresponding normal DNA exhibited such methylation. Subsequently, we examined whether aberrant methylation could be detected in corresponding serum DNA, and found that 13 of 44 (30%) patients with p16 promoter methylation in tumor DNA demonstrated abnormal methylation in their serum DNA. Moreover, abnormal methylation was found in the serum of patients in all clinical stages, suggesting that early colorectal cancer could be detected using the MSP method.

Quantitative detection of CEA expressing free tumor cells in the peripheral blood of colorectal cancer patients during surgery with real-time RT-PCR on a LightCycler.

We applied novel real-time reverse transcriptase-polymerase chain reaction (RT-PCR) with a LightCycler for quantitative detection of carcinoembryonic antigen (CEA) mRNA expressing tumor cells in the peripheral blood of colorectal cancer patients. Analysis of peripheral blood samples from 99 potentially curative colorectal cancer patients revealed a significantly higher mean CEA mRNA value in post-operative bloods (18.71) than in pre-operative blood (1.03) (P=0.003). Kaplan-Meier analysis demonstrated disease free survival of patients with positive CEA mRNA in post-operative blood to be significantly shorter than in cases negative for CEA mRNA (P=0.03). These results suggest that tumor cells could be shed into the bloodstream during surgical procedures, and these free tumor cells are accompanied by a poor patient outcome. Real-time quantitative RT-PCR is a useful technique for quantitative assessment of free tumor cells in the peripheral blood of colorectal cancer patients.

Laparoscopy-assisted distal gastrectomy with systemic lymph node dissection for early gastric carcinoma: a review of 43 cases.

BACKGROUND: Recently, laparoscopy and laparoscopy-assisted surgery have been used increasingly as less-invasive alternatives to conventional open surgery. But the use of this approach in gastric carcinoma has received little attention, possibly from the low incidence of early-stage disease in the West and the relative complexity of the surgical procedure. STUDY DESIGN: A prospective feasibility study of laparoscopy-assisted distal gastrectomy was performed in patients with histologically confirmed gastric carcinoma located in the lower or middle third of the stomach. Patients whose preoperative evaluations, including endoscopic ultrasonography and computerized tomography, led to a diagnosis of T1 N0 stage disease, and who had no advanced disease discovered during laparoscopy, were eligible. Intraoperative blood loss, time of operation, mortality, and morbidity were assessed, along with the number of lymph nodes retrieved and shortterm survival. RESULTS: Between 1998 and 1999, 43 patients were enrolled. Laparoscopy-assisted distal gastrectomy was converted to an open procedure in one patient. There were no operative or in-hospital deaths, but the incidence of anastomotic leakage was 14% (6 of 43). The mean blood loss was 239 mL, the time of operation was 225 minutes, and lymph node retrieval was 20.2 nodes. These results are comparable with a series of conventional open operations. One patient died of recurrent disease, and all other patients remain disease-free to date. Port-site recurrence was not observed. CONCLUSIONS: Although laparoscopy-assisted distal gastrectomy was equivalent to open surgery in several clinical parameters, the relatively high morbidity was a drawback. Its appropriateness to gastric cancer surgery must be verified by further studies.

Effect of transfection with human interferon-beta gene entrapped in cationic multilamellar liposomes in combination with 5-fluorouracil on the growth of human esophageal cancer cells in vitro.

When human esophageal cancer cells were transfected with the human interferon-beta (hIFN-beta) gene entrapped in cationic multilamellar liposomes, the growth of all cancer cells tested was suppressed in a dose-dependent manner. The 50% inhibitory concentration (IC50) of the hIFN-beta gene entrapped in the liposomes ranged from 16 to 176 ng plasmid DNA/ml culture medium. Among the 10 cell lines examined, NUEC3, NUEC4, TE-3 and WSSC cell lines were highly susceptible to transfection with this gene entrapped in the liposomes. The IC50 values of the hIFN-beta gene entrapped in the liposomes with respect to cell growth were positively-correlated with those of exogenous cytokine hIFN-beta, suggesting that the antiproliferative effect of hIFN-beta gene entrapped in the liposomes can be mainly ascribed to the function of hIFN-beta produced by cells transfected with the gene. Two days after transfection with the liposome-entrapped gene, the concentration of hIFN-beta secreted into the medium was determined. Even though the level of hIFN-beta observed in the medium was lower than that of the IC50 of exogenously added hIFN-beta, the inhibitory potency of the hIFN-beta gene entrapped in the liposomes on the cell growth was remarkable. When the esophageal cancer cells were treated with 5-fluorouracil (5-FU) in the presence of a low concentration of liposome-entrapped-gene, the rate of growth inhibition of these cells increased over that caused by either 5-FU or hIFN-beta gene entrapped in the liposomes alone. All these data suggest that combination therapy with the hIFN-beta gene entrapped in cationic multilamellar liposomes and the anticancer drug 5-FU would be beneficial for preoperative treatment of carcinoma of the esophagus.

PGP9.5 overexpression in esophageal squamous cell carcinoma.

BACKGROUND/AIMS: PGP9.5 is a ubiquitin hydrolase widely expressed in neuronal tissue at all stages of neuronal differentiation and has been used as a neuroendocrine marker. Recently, it has been proved that PGP9.5 expression was highly observed in squamous cell carcinoma of lung cancer, suggesting that it might be a tumor marker for squamous cell carcinoma. To better characterize its role in digestive tract cancers, we examined PGP9.5 expression retrospectively in esophageal cancers. METHODOLOGY: We examined PGP9.5 expression retrospectively in 40 resected esophageal cancers (squamous cell carcinoma) and 10 gastric cancers (adenocarcinoma) using immunohistochemistry. RESULTS: Of 40 esophageal cancer specimens, 19 (48%) exhibited positive staining with PGP9.5 in most tumor cells, while no PGP9.5 expression was observed in any of the 10 gastric cancers. CONCLUSIONS: Although the precise mechanism underlying the effect of PGP9.5 on oncogenicity remains to be proven, it was confirmed that it may be a potential marker for esophageal squamous cell carcinoma.

Phosphorylation of inositol 1,4,5-triphosphate receptor 1 during in vitro maturation of porcine oocytes.

During fertilization in mammalian species, a sperm-induced intracellular Ca(2+) signal ([Ca(2+)](i)) mediates both exit of meiosis and oocyte activation. Recently, we demonstrated in mouse oocytes that the phosphorylation levels of inositol 1,4,5 trisphosphate receptor type1 (IP(3)R1), the channel responsible for Ca(2+) release and oscillations during fertilization, changed during maturation and fertilization. Therefore, we examined the expression and phosphorylation of IP(3)R1 during in vitro maturation of pig oocytes. Here, our present study shows that expression of IP(3)R1 protein did not change during maturation, although the phosphorylation status of the receptor, specifically at an MPM-2 epitope, did. We found that while at the beginning of maturation IP(3)R1 lacked MPM-2 immunoreactivity, it became MPM-2 reactive by 24 h and reached maximal reactivity by 36 h. Interestingly, the acquisition of MPM-2 reactivity coincided with the activation of p34(cdc2) kinase and mitogen-activated protein kinase (MAPK), which are involved in meiotic progression. Following completion of maturation, inactivation of MAPK by U0126 did not affect IP(3)R1 phosphorylation, although inactivation of p34(cdc2) kinase by roscovitine dramatically reduced IP(3)R1 phosphorylation. Neither inhibitor affected total expression of IP(3)R1. Altogether, our results show that IP(3)R1 undergoes dynamic phosphorylation during maturation and this might underlie the generation of oscillations at fertilization.

Evaluation of the histoculture drug response assay as a sensitivity test for anticancer agents.

PURPOSE: To investigate whether the effects of anticancer agents are able to be predicted, the results of sensitivity tests on anticancer agents were compared with the results of preoperative chemotherapy. METHODS: Biopsies were taken from 25 patients with esophageal cancer and 10 with gastric cancer, before chemotherapy, and the drug sensitivity was determined after culturing for 1 week. The chemotherapy consisted of low-dose cis-diamino-dichloroplatinum + 5-fluorouracil, and its clinical effect was determined after 3 or 5 weeks. RESULTS: Sensitivity was determined in 20 of the esophageal cancer patients and 8 of the gastric cancer patients, accounting for 80% of all the patients. Of the 11 patients judged to have sensitivity by the histoculture drug response assay (HDRA), 7 had a partial response, and of the 17 judged to have no sensitivity, 16 had a minor response or no change (NC). It was thus demonstrated that predictions of the effect of anticancer agents could be made with considerable accuracy using HDRA. CONCLUSION: The prognosis of the NC patients was poor, and distant metastasis, thought to be an adverse effect, soon appeared. From the viewpoint of both medical costs and patient quality of life, treatments other than preoperative chemotherapy should be selected for patients assessed to have NC. We believe that these sensitivity tests should be introduced clinically.

Colorectal cancers with both p16 and p14 methylation show invasive characteristics.

Recent studies indicated that p16 and p14 inactivation owing to promoter methylation was important for colorectal tumorigenesis. In this study, we examined the methylation status of these genes in 86 primary colorectal cancers using methylation-specific PCR (MSP) and correlated the results with the clinicopathological features of the patients. Aberrant promoter methylation of p16 and p14 genes was detected in 43 of 86 (50%) and 25 of 86 (29%) colorectal cancers, respectively. Next, we examined the correlation of methylation status with the clinicopathological features. We found a significant difference in maximal tumor size (P=0.022) when patients with both p16 and p14 methylation were compared to other patients. On the other hand, there was no significant difference in other factors, such as the extent of tumor and Dukes stage. These results suggested that colorectal cancer with both p16 and p14 methylation has the same invasiveness at a smaller size compared to that of the cancer with neither p16 nor p14 methylation. Inactivation of both p16 and p14 genes may result in a malignant change in colorectal cancer cells, leading to advanced cancers with a smaller size than those with p16 or p14 activity.

Modification of adriamycin pharmacokinetics by direct electric current in rats.

Adriamycin (ADR, doxorubicin), a drug having cardiotoxicity, is electrically charged as a cation in blood. We therefore investigated whether iontophoresis caused by direct electric current (DC; 50 microA, 90 min) would cause systemic modification of ADR pharmacokinetics. Cathode and anode were placed into a right kidney and muscles of the abdominal wall, respectively, in six Donryu rats. Urinary excretion of ADR, as measured by catheterizing into the right kidney, was significantly higher in the DC group than in the controls (p < 0.05). Both plasmic and renal ADR clearances were significantly higher in the DC group (p<0.005 and p<0.001, respectively). Tissue ADR concentrations were significantly lower in the DC group (heart: p<0.003; liver and lung: both p<0.05). These results suggest that electric therapy might potentially induce modification of ADR pharmacokinetics by iontophoresis, and that the therapy might effectively change ADR concentration both locally and systemically.

Laparoscopic distal gastrectomy for severe corrosive gastritis: report of a case.

We report a case of severe corrosive gastritis caused by alkali ingestion, which was successfully treated by laparoscopic gastrectomy. A 38-year-old Japanese woman attempted suicide by ingesting Drano. She was treated conservatively for 3 months, but severe stenosis of the antrum secondary to scarification resulted in outlet obstruction. A Billroth I distal gastrectomy was performed through a laparoscopy-assisted approach. The patient had an uneventful postoperative course and was discharged on the 13th day after her operation. She was subsequently able to tolerate a normal diet, gained weight, and is now emotionally stable.

Molecular detection of p16 promoter methylation in the serum of recurrent colorectal cancer patients.

We previously proved that p16 promoter methylation present in the tumors of colorectal cancer patients can be detected in the serum of those same patients using methylation-specific PCR (MSP). To seek the possibility that this technique could be applied to the monitoring of cancer recurrence, we examined the p16 methylation using MSP. We detected tumor DNA in the serum of 31 of 45 (69%) patients with recurrent colorectal cancer. No methylation was found in serum DNA of 50 patients with colorectal cancers whose corresponding tumor DNA had no methylation in p16 promoter. These results suggested that MSP might be a sensitive and useful method to detect recurrent colorectal cancer in serum.

Phase II study of paclitaxel with 3-h infusion in patients with advanced gastric cancer.

BACKGROUND: To increase the options for agents for gastric cancer chemotherapy, we performed a phase II clinical trial on the use of a 3-h infusion of paclitaxel to confirm its efficacy and the feasibility of its use in patients with advanced gastric cancer. METHODS: Thirty-two (32) patients with measurable metastatic gastric cancer were enrolled in this study. Seventeen patients (53%) had received prior chemotherapy for metastatic disease, 4 patients (13%) had adjuvant chemotherapy alone, and 11 patients (34%) were chemotherapy-naive. Paclitaxel was intravenously infused for 3 h, at a dose of 210 mg/m(2), once every 3 weeks. To prevent hypersensitivity reactions, standard premedication was administered to all patients. RESULTS: Nine (28%; 9/32 ) objective partial responses (PRs) were observed (95% confidence interval [CI], 14%-47%), and the remaining 23 patients showed stable (12 patients; 37.5%) and progressive disease (11 patients; 34.4%). The median time to response was 20 days (range, 14-38 days). The median response duration was 87 days (range, 50-103 days). The median survival of all patients was 234 days (range, 13-646+ days). The major adverse reactions were myelosuppression (grade 3/4 leukopenia and neutropenia were observed in 59% and 88% of the patients, respectively), alopecia, and peripheral neuropathy. Peripheral neuropathy was observed in 19 patients, however, most of the patients recovered after the completion of treatment. CONCLUSION: A 3-h infusion of paclitaxel is an effective therapy for advanced gastric cancer and is clinically well tolerated by the patients.