Ceramide induces apoptosis in human lung adenocarcinoma A549 cells through mitogen-activated protein kinases.

AIM: To provide experimental data for further research on the signal transduction of apoptosis in lung adenocarcinoma cells, we examined the effects of exogenous C2-ceramide administration on several members of the mitogen-activated protein kinase (MAPK) superfamily and caspase-3 in A549 cells. METHODS: Cell viability and apoptosis were analyzed by cell counting kit-8 assay and flow cytometry. Various MAPK and caspase-3 proteins were detected by Western blotting. RESULTS: C2-ceramide selectively altered the phosphorylation state of members of the MAPK superfamily, causing hyperphosphorylation of mitogen-activated protein kinase kinase (MEK)1/2 and the p38 MAPK, but not affecting the phosphorylation of extracellular signal-regulated kinase 1/2 and the c-Jun N-terminal kinase. SB-203580 (a p38 MAPK inhibitor) and p38 siRNA, but not U0126 (a MEK inhibitor), partially rescued cell death induced by C2-ceramide. C2-ceramide promoted the activation of caspase-3. CONCLUSION: Exogenous C2-ceramide induced apoptosis in human lung adenocarcinoma A549 cells. The activation of MAPK and caspase-3 were involved in the mechanisms of C2-ceramide-induced apoptosis in A549 cells.

Long-term effects of subcutaneously injected 2,3,7,8-tetrachlorodibenzo-p-dioxin on the liver of rhesus monkeys.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) accumulates and remains stable in the fatty tissues and liver of rodents for a long time. Considering the pronounced difference between species, long-term, low dose hepatic effects of TCDD were investigated after subcutaneous administration of TCDD into rhesus monkeys during pregnancy. Macroscopic and histopathological examination of the liver carried out 4 y after TCDD administration demonstrated intrahepatic focal fatty changes, infarction, hemorrhage, microthrombi-formation, sinusoidal ectasia, small hepatocyte hyperplasia, and increased number of alpha-smooth muscle actin (alpha-SMA)-positive cells. An electron microscopic study disclosed sinusoidal endothelial cell degeneration and injury in the liver of TCDD-treated monkeys. Western blot analysis showed downregulation of aryl hydrocarbon receptor (AhR) protein expression and decreased level of vascular endothelial (VE) cadherin but increased expression levels of CYP1A1 and transforming growth factor beta (TGF-beta) protein in the liver tissues. These changes observed in TCDD-exposed monkeys indicated sinusoidal endothelial cell injury and impairment in intrasinusoidal microcirculation. Infarction, focal fatty change, and microthrombi-formation are considered to be closely associated with intrahepatic circulatory impairment. Increased number of alpha-SMA-positive cells and decreased level of VE cadherin expression in the liver tissues might also be associated with sinusoidal endothelial cell injury. In addition, downregulation of AhR expression and increased CYP1A1 protein levels in the liver were consistent with persistent effects of TCDD. Although it has been reported that TCDD induced endothelial cell injury, this is the first report to describe vascular disorders and protein expression in the liver after injection with TCDD in a primate model.

Ornithine decarboxylase, mitogen-activated protein kinase and matrix metalloproteinase-2 expressions in human colon tumors.

AIM: To investigate the expressions of ornithine decarboxylase (ODC), MMP-2, and Erk, and their relationship in human colon tumors. METHODS: ODC activity, MMP-2 expression, and mitogen-activated protein (MAP) kinase activity (Erk phosphorylation) were determined in 58 surgically removed human colon tumors and their adjacent normal tissues, using [1-14C]-ornithine as a substrate, ELISA assay, and Western blotting, respectively. RESULTS: ODC activity, MMP-2 expression, and Erk phosphorylation were significantly elevated in colon tumors, compared to those in adjacent normal tissues. A significant correlation was observed between ODC activities and MMP-2 levels. CONCLUSION: This is the first report showing a significant correlation between ODC activities and MMP-2 levels in human colon tumors. As MMP-2 is involved in cancer invasion and metastasis, and colon cancer overexpresses ODC, suppression of ODC expression may be a rational approach to treat colon cancer which overexpresses ODC.

Serum levels of tissue inhibitor of metalloproteinases-1 (TIMP-1) in colorectal cancer patients.

PURPOSE: Expression of tissue inhibitor of metalloproteinases (TIMP)-1 in colorectal cancer tissue is known to be related to disease progression; however, the clinical significance of measuring the blood level of TIMP-1, which we evaluate herein, has not yet been clarified. METHODS: The serum level of TIMP-1 was measured by a one-step enzyme immunoassay in 123 patients who underwent resection of primary colorectal cancer. RESULTS: An elevated level of serum TIMP-1 was associated with advanced Dukes' stage ( P = 0.03), greater diameter of the primary tumor ( P = 0.03), more lymph node metastasis ( P = 0.04), and liver metastasis ( P < 0.001). There was a weakly positive correlation between the serum carcinoembryonic antigen (CEA) level and the serum TIMP-1 level. In patients who underwent potentially curative resection, the disease-free survival was not different between those with a high TIMP-1 level (>=203.5 ng/ml, n = 32) and those with a low TIMP-1 level (<203.5 ng/ml, n = 66, P = 0.62). In patients with Dukes' stage D cancer who underwent noncurative resection, the survival times were not different between those with a high TIMP-1 level ( n = 13) and those with a low TIMP-1 level ( n = 10, P = 0.20). CONCLUSIONS: Elevated levels of serum TIMP-1 reflect the extent of colorectal cancer, without a close correlation with the serum CEA level. These findings suggest that measuring the serum TIMP-1 level would not help to predict the prognosis of patients with colorectal cancer.

[Results of prophylactic hepatic arterial chemotherapy for liver metastases of Dukes C colorectal cancer--correlation with tumoral expression of dihydropyrimidine dehydrogenase, thymidylate synthase, p53, or orotate phosphoribosyl transferase].

The purpose of this study was (1) to disclose data from a non-randomized trial of prophylactic hepatic arterial chemotherapy for liver metastases from Dukes'C colorectal cancer, (2) to examine the influence of the expression of dihydropyrimidine dehydrogenase (DPD), thymidylate synthase (TS), and p53 in the primary lesion on this chemotherapy, and (3) to examine the expression of orotate phosphoribosyl transferase (OPRT) mRNA levels in the cases of recurrence included in this study. Patients who underwent curative resection of Dukes'C colorectal cancer between November 1996 and April 2000 were examined. After curative resection, patients were non-randomly divided into two groups after obtaining their informed consent: Hepatic arterial infusion (HAI) group patients (n = 28) were given 5-FU (500 mg/body for 1 h per week, repeated 50 times) via the hepatic artery and peroral UFT-E after resection of Dukes'C colorectal cancer. Control group patients (n = 21) received UFT-E alone. Liver metastasis-free survival did not differ between the groups. Immunohistochemical examinations revealed that the expression of tumoral DPD or p53 was unlikely to affect the hepatic recurrence, although patients with a low expression of TS tended to have better survival in both groups. However, multivariate analysis by the Cox proportional hazard model revealed that a significant prognostic factor influencing the hepatic recurrence is extensive venous invasion. Expression levels of OPRT mRNA, measured in tumors of patients with recurrence (n = 6 for the HAI group; and n = 4 for the control group) were not significantly different between the groups. These results suggest that (1) intermittent hepatic arterial infusion of 5-FU in addition to oral UFT-E was not more useful than administration of UFT alone, and (2) the expression of DPD, TS, p53, and OPRT in the primary lesion was unlikely to affect the prognosis of patients included in this study.

Determining the levels of matrix metalloproteinase-9 in portal and peripheral blood is useful for predicting liver metastasis of colorectal cancer.

BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is one of the MMPs that play an important role in cancer invasion and metastasis. Increased levels of MMP-9 in tumor tissue have been found to correlate with advanced stages of colorectal cancer. However, the clinical significance of determining the levels of MMP-9 in blood samples from patients with colorectal cancer has not yet been clarified. The purpose of this study was to clarify the relationship between the clinicopathological variables of colorectal cancer and MMP-9 levels of drainage (portal) or peripheral venous blood and to examine whether this assay would be useful for predicting liver metastasis. METHODS: Blood samples were obtained from peripheral and drainage veins of 102 patients with colorectal cancer during surgery and the plasma levels of MMP-9 were determined by a one-step sandwich enzyme immunoassay. RESULTS: The levels of portal MMP-9 were significantly higher than those of peripheral blood (P < 0.01, n = 102). The levels of MMP-9 in peripheral venous blood did not correlate with any of the 12 clinicopathological variables examined, while the levels of MMP-9 in portal blood correlated with macroscopic type of the primary tumor (P = 0.02), Dukes' stage (P = 0.03), liver metastasis (P < 0.01) and lymph node metastasis (P = 0.02). By setting the cutoff ratio of portal to peripheral MMP-9 levels at 1.6 in patients with curative resection (n = 73), elevated ratios predicted subsequent emergence of liver metastases with 77.8% sensitivity, 81.3% specificity and 80.8% accuracy. CONCLUSION: The results suggest that synchronous determination of the levels of MMP-9 in portal and peripheral blood would be useful for selecting colorectal cancer patients at high risk of hepatic recurrence.

Muscle strength and walking ability after laparoscopic hernioplasty versus conventional repair.

PURPOSE: We examined muscle strength and walking ability after hernia repair to compare the effects of laparoscopic hernioplasty and conventional repair. METHODS: Twenty-seven patients with primary inguinal hernias were randomly divided into two groups according to the surgical procedure: 15 patients were treated with laparoscopic hernioplasty and 12 with conventional repair. Two types of muscle testing around the inguinal region, one by manual examination and one using a musculator, were done preoperatively and 1 week postoperatively. Walking exercise tests were performed at the same time as muscle testing. RESULTS: Manual examination showed that the postoperative muscle strength of the iliopsoas muscle was decreased in six patients from the conventional repair group, but not in any of those from the laparoscopic hernioplasty group. The musculator showed decreased strength of the iliopsoas muscle after conventional repair, but not after laparoscopic hernioplasty. The walking exercise test showed that conventional hernia repair influenced walking. CONCLUSION: A decline in muscle strength of the iliopsoas muscle and walking ability was evident after conventional repair, but not after laparoscopic hernioplasty. Thus, laparoscopic hernioplasty is superior to conventional repair from the perspective of muscle testing and walking ability.

Fulminant amoebic colitis with perforation successfully treated by staged surgery: a case report.

Radical surgery for fulminant amoebic colitis leads to extremely high mortality; however, resective surgery is mandatory if a patient develops massive fecal peritonitis. We herein report an extremely rare case of fulminant amoebic colitis with multiple perforations, which was successfully treated by staged surgical procedures. A 48-year-old man who had been treated with predonisolone under a diagnosis of ulcerative colitis was admitted. Biopsy specimens from the colonic mucosa revealed Entamoeba histolytica. On the day of diagnosis, he developed severe abdominal pain and underwent emergency laparoptomy, showing total colonic gangrene with multiple perforations associated with massive fecal peritonitis. Subtotal colectomy, mucous fistula of the rectosigmoid, and ileostomy were performed. He recovered well although disseminated intravascular coagulopathy developed postoperatively. As the middle and upper part of rectum was found to be severely stenotic 4 months after surgery, we performed proctectomy, ileal pouch anal canal anastomosis, and diverting ileostomy, which was reversed 6 months later. The patient has been well with satisfactory anal function 37 months after the initial surgery. This case suggests that (1). early and accurate diagnosis of amoebiasis is important to avoid surgical intervention, and (2). staged surgery including total colectomy should be considered as one of the treatment choices even in patients with total necrotizing amoebic colitis.

[A case of postoperative liver metastasis of alpha-fetoprotein producing gastric cancer successfully treated with intrahepatic chemotherapy (adriamycin, cisplatin, irinotecan hydrochloride)].

There are only a few reports demonstrating effective regimens for AFP-producing gastric cancer. We report a case of alpha-fetoprotein (AFP)-producing gastric cancer successfully treated with postoperative intrahepatic chemotherapy, together with a review of the literature. A 63-year-old man was diagnosed as having multiple liver metastases 6 months following total gastrectomy for type 2 gastric cancer. Serum AFP level was markedly elevated at 291.4 ng/ml although the level was not determined at the time of gastrectomy. Twenty cycles of intrahepatic arterial infusion with adriamycin, cisplatin, and irinotecan hydrochloride resulted in a 93% decrease in the liver metastases along with normalization of the serum AFP level. These results suggest that this regimen is worth trying for patients with liver metastasis from AFP-producing gastric cancer.

[Hepatic recurrence after prophylactic hepatic arterial infusion of 5-fluorouracil for Dukes' C colorectal cancer--correlation with the expression of dihydropyrimidine dehydrogenase in the primary tumor].

PURPOSE: The purpose of this study was twofold: (1) to disclose the intermediate outcome of a non-randomized trial of prophylactic hepatic arterial infusion chemotherapy (PHAI) for curatively resected Dukes' C colorectal cancer performed between November 1996 and April 2000, and (2) to examine the relationship between the expression of dihydropyrimidine dehydrogenase (DPD) in tumor tissue and the efficacy of this chemotherapy. PATIENTS AND METHODS: The oncological outcomes were compared between patients (n = 28) receiving PHAI (5-FU: 500 mg/body/w x 50 cycles) plus oral administration of UFT-E (400 mg/body/day, for 24 months) and those (n = 21) receiving UFT-E alone. The levels of tumoral DPD were determined in a total of 43 patients (n = 25, PHAI group; n = 18, control group) by an enzyme-linked immunosorbent assay. RESULTS: Seven (25%) in the PHAI group and four (19%) in the control group developed liver metastasis postoperatively. The liver metastasis-free survival was not different between the groups (p = 0.94). When the analysis was restricted to patients who developed liver metastasis, the duration from surgery to detecting liver metastasis tended to be longer in the PHAI group (p = 0.09). In addition, the overall survival tended to be better in the PHAI group (p = 0.12). In the control group, the level of DPD was higher in patients who developed liver metastases (n = 4) than in those who did not (n = 14, p = 0.04). However, in the PHAI group, the level of DPD was not different regardless of the occurrence of liver metastases (p = 0.30). CONCLUSIONS: These results suggest that (1) PHAI is unlikely to improve the prognosis of Dukes' C patients remarkably, and (2) the efficacy of this regimen cannot be predicted by determining the levels of tumoral DPD.

[Short-term intraperitoneal infusion of cisplatin and OK-432 for a case of malignant mesothelioma of the peritoneum].

A case of malignant mesothelioma of the peritoneum successfully treated with short-term intraperitoneal chemotherapy is reported. A 67-year-old woman underwent laparotomy for ascending colon cancer. During, laparotomy, numerous mucoid nodules were found on the peritoneal surface, even though the colonic tumor did not invade the serosa. Ileocecal resection (D1) with partial omentectomy was performed. Intraperitoneal infusion with cisplatin (200 mg) was started immediately after surgery and additional cisplatin (150 mg) was administered with OK-432 (30KE). The serum level of CA125 rapidly decreased after the chemotherapy, and was normalized 3 months postoperatively. The histological diagnosis of the peritoneal lesions was malignant mesothelioma of the peritoneum (diffuse type). The patient is living without any evidence of recurrence 10 months postoperatively. These results suggest that this short-term chemotherapy is worth trying in cases of malignant mesothelioma of the peritoneum.

Primary thyroid carcinoma found with esophageal stenosis: report of a case.

The typical symptoms of primary thyroid cancer are a cervical mass and cervical lymphadenopathy, while dyspnea, hoarseness, or dysphagia can occur in the presence of extrathyroidal involvement. Esophageal involvement or stenosis causing dysphagia without any influence on the trachea is rare because of the anatomical location of the esophagus and the trachea. We report herein a case of primary thyroid carcinoma advancing behind the esophagus with the trachea intact, which was difficult to diagnose by esophagoscopy or roentgenogram. Thus, thyroid tumors need to be differentiated from other possible causes of esophageal stenosis. Computed tomography, magnetic resonance imaging, and ultrasonography are useful diagnostic modalities to detect thyroid tumors causing esophageal stenosis.

Risk of missing colorectal cancer during laparoscopic cholecystectomy.

PURPOSE: The increased number of patients undergoing laparoscopic cholecystectomy (LC) is associated with a risk of missing concomitant colorectal cancers; however, the incidence and cause have not yet been well recognized. Our aim, therefore, was to evaluate these factors. METHODS: This retrospective study evaluated data on 473 patients with benign gallbladder diseases, who underwent LC between January 1991 and December 1999. Among these 473 patients, 2 (0.4%) were thought to have had detectable cancer at LC. RESULTS: The first patient was a 59-year-old woman who underwent palliative resection for ascending colon cancer associated with liver and pulmonary metastases 10 months following LC when laboratory data showed a low hemoglobin level (10.0 g/dl). The other patient, a 50-year-old man, underwent resection for Dukes' C sigmoid colon cancer 6 months following LC. At LC, the patient did not present with any symptoms suggesting the existence of colorectal cancer and the laboratory data were normal. CONCLUSIONS: These results indicate that although an extremely low incidence of missed colorectal cancers does not justify routine screening for colorectal cancer before LC in terms of cost-effectiveness, careful attention to preoperative physical findings and laboratory data as well as meticulous techniques and full diagnostic visualization of the large-bowel intraoperatively may reduce the potential risk of missing coexisting colorectal cancers during LC.

Influence of perioperative blood transfusion on the prognosis of patients with gastric cancer receiving anticancer chemotherapy.

BACKGROUND: The deleterious effect of blood transfusions on survival has been reported in patients with cancers of various organs. However, it remains unclear whether there is any adverse effect of blood transfusion when the patients are administered anticancer drugs after surgery for gastric cancers.METHODS: Data from patients with gastric resection for advanced gastric cancer were retrospectively analyzed to determine the influence of perioperative blood transfusion on the survival rate. All patients were administered anticancer drugs (mitomycin C [MMC] and tegafur-uracil [UFT]). Sixty-nine (33%) of 208 patients received blood transfusion perioperatively, while 139 patients (67%) did not receive transfusion. Multivariate analysis of clinicopathologic prognostic factors, including blood transfusion, was performed. Lymphocyte subsets were measured to investigate the immunosuppressive effect of blood transfusion.RESULTS: The 5-year survival rate was 48.8% in the 69 transfused patients and 66.9% in the 139 non-transfused patients ( P < 0.01). Cox's multiple regression analysis showed that, when patients received anticancer drugs, perioperative blood transfusion was not a significant factor affecting survival after the gastric cancer surgery. However, the CD4/CD8 ratio at 3 months after the surgery was significantly lower in the transfused group than in the non-transfused group.CONCLUSION: Blood transfusion did not affect the survival of operated patients who received postoperative adjuvant chemotherapy. However, the finding that the ratio of CD4/CD8 after surgery was significantly higher in the non-transfused group than in the transfused group supports the notion that transfusion causes broad-spectrum immunosuppression.