Expression of cyclin D1, CDK4 and p27KIP1 is associated with the p16MTS1 gene status in human esophageal carcinoma cell lines.

p16MTS1/INK4A negatively regulates cell cycle progression by inhibiting the cyclin D/CDK4 complex that phosphorylates pRb. Frequent homozygous deletions of the p16 gene were recently found in various tumor cell lines. We examined the relationship between the genetic status of p16 and the expression of the cell cycle regulating molecules in human esophageal carcinoma cell lines. Out of eight human esophageal carcinoma cell lines, seven (67.5%) and six (75%) cell lines showed homozygous deletions of the p16 and p15 genes, respectively. All the p16-negative cell lines expressed high levels of cyclin D1, CDK4 and p27KIP1 proteins. Interestingly, the expression level of cyclin D1 was closely correlated to the levels of not only CDK4 but also p27KIP1 protein in p16-negative cell lines. Furthermore, all the p16-negative cell lines expressed Rb protein of approx 110 kDa which corresponds to the phosphorylated form, whereas the cell line with intact p15 and p16 genes did not express pRb. These results suggest that the expression of cyclin D1, CDK4, Rb and p27 is associated with the p16 gene status in esophageal carcinoma cell lines. Alternatively, loss of the p16 gene and subsequent over-expression of cyclin D1 and CDK4 might be involved in autonomous growth of esophageal carcinoma cells.

[Low-dose FP (5-FU+low-dose CDDP) therapy for far advanced upper gastro-intestinal carcinoma].

Low-dose FP therapy was undertaken in 25 patients with far advanced or recurrent carcinoma (13 stomach, 7 esophagus, 3 colon, 1 gallbladder, 1 pancreas). This therapy consisted of intermittent infusion of CDDP (10 mg/body X 5 days) and continuous infusion of 5-FU (500 mg/body X 5 days) for 5 days with 2-day intervals. Patients were treated with at least 2 courses of low-dose FP therapy. Of the 25 patients, 12 with esophageal (7) or gastric (5) carcinoma, in whom curative resection was considered impossible before the operation, were subjected to neoadjuvant chemotherapy. The response rate in the neoadjuvant therapy was 100% (2 disappeared and 5 decreased in size) in the esophageal cancer and 60% (3 decreased in size) in the gastric cancer. But, in 6 patients with esophageal cancer, radiotherapy was combined. In the neoadjuvant cases, pathological effect of Grade 2 was noted in 3 of the 7 esophageal cancers and 1 of the 5 gastric cancers. Of the remaining 13 unresectable patients, a significant improvement of performance status was found in 6 patients. In 19 patients treated with low-dose FP therapy only, leukopenia of Grade 3 was not observed, and there was no nephrotoxicity. Low-dose FP therapy is safe and useful as a neoadjuvant chemotherapy for patients with far advanced esophageal or gastric cancer.

Xanthogranulomatous cholecystis. Cell composition and a possible pathogenetic role of cell-mediated immunity.

Thirty-three cases of xanthogranulomatous cholecystitis (XGC) exhibiting the typical morphologic features were studied by light and electron microscopy and immunohistochemical techniques. Incidence of XGC was 4.2% of the surgically resected gallbladder diseases. Histologically, the granulomatous lesion of XGC principally consisted of accumulations of foam cells and lymphocytes. Variable numbers of multinucleated giant cells, granulocytes and fibroblastic cells were also noted. With respect to the origin of foam cells, it was considered that the vast majority of foam cells were derived from monocytes/macrophages because they were invariably positive for KP1, HAM56, CD11b and CD68. Interspersed among macrophage foam cells, many T lymphocytes were identified. The subtyping of T cells indicated a heterogenous population composed of both CD4+ and CD8+ lymphocytes typically in a ratio of 1:2. Macrophages and T lymphocytes demonstrated a marked expression of HLA-DR antigen. Electron microscopic and immunohistochemical double-staining observation demonstrated intimate apposition of T lymphocytes to macrophages or macrophage foam cells. The results indicate that XGC is a granulomatous disorder characterized by accumulations of macrophage foam cells and T cells. Delayed type hypersensitivity reaction of cell-mediated immunity may be implicated in the pathogenesis of XGC.

Biomodulation by hyperthermia of topoisomerase II-targeting drugs in human colorectal cancer cells.

We examined whether heat stress could enhance the sensitivity of human colon cancer WiDr cells to topoisomerase II-targeting anticancer agents, etoposide (VP-16) and teniposide (VM-26), and also determined the most effective timing for the drug administration after exposure to hyperthermia. Both topoisomerase II contents and topoisomerase II activity were significantly increased in WiDr cells 3 to 12 h after heat stress at 43 degrees C for 1 h, in comparison with those immediately after the heat stress. Cytotoxicity by VP-16 was most significantly enhanced 3 to 12 h after exposure to 43 degrees C for 1 h, but no synergistic effect was observed when the drug was administered immediately after the heat stress. A combination of VM-26 with heat stress, but not that of a topoisomerase I-targeting camptothecin derivative (CPT-11), or vincristine, showed a synergistic cytotoxic effect on WiDr cells. VP-16 alone induced cellular accumulation at the G2 + M phase, whereas the combination of VP-16 and heat stress further increased the cell population at the G2 + M phase, and decreased S-phase cells. A possible application of the combination of VP-16 and hyperthermia in clinical use is discussed.

Snake-strike--induced ischemic colitis with colonic stricture complicated by disseminated intravascular coagulation.

A 57-year-old farmer was struck on the right thumb by a pit viper (Agkistrodon blomhoffü). Subsequently, he had acute renal failure and disseminated intravascular coagulation (DIC), associated with melanotic stools and abdominal pain. Renal failure caused by renal cortical necrosis was successfully treated with hemodialysis. A double-contrast barium enema examination revealed multiple stenoses of the colon, regional edema, and longitudinal ulcer. Histologic examination of the stenotic lesions after laparotomy revealed fibrosis of both submucosa and proper muscle layer, with fibrotic thickness in the small arteries of the colonic wall, indicating that ischemic colitis was associated with DIC. In this case, DIC from viper toxins played an etiologic role in the development of ischemic colitis with stricture, as well as acute renal failure.

Prevention of recurrent bleeding from gastric ulcer with a nonbleeding visible vessel by endoscopic injection of absolute ethanol: a prospective, controlled trial.

We performed a prospective, randomized trial to assess the efficacy of endoscopic injection therapy with absolute ethanol in preventing recurrent bleeding in patients with nonbleeding visible vessels in gastric ulcers. During the period of 1990 to 1993, 62 patients who bled were found to have gastric ulcers with nonbleeding visible vessels; all of them were enrolled for this trial. The 62 patients were randomly divided into two groups, which were comparable at entry. In group I (33 patients), we performed endoscopic injection therapy with absolute ethanol. In group II (29 patients), we sprayed the ulcers with 0.1% epinephrine and thrombin. Endoscopic injection therapy with ethanol was performed at the second endoscopy in the patients in both groups who had recurrent bleeding. Among the 33 patients in group I, 4 patients (12.1%) rebled after the initial ethanol injection therapy, whereas 10 of 29 patients (34.5%) rebled in the control group (p < .05). No patients in group I required surgical intervention, and ultimate hemostasis was achieved in all 33 group I patients (100%), indicating that endoscopic ethanol injection therapy achieves ultimate hemostasis and prevents recurrent bleeding in patients with gastric ulcers and nonbleeding visible vessels.

Concurrent amplification of cyclin E and CDK2 genes in colorectal carcinomas.

The genetic changes of cyclin A, DI, E and CDK2 were examined in human colorectal carcinomas by Southern-blot analysis. Gene amplification of cyclin E was detected in 5 of 53 (9.4%) primary colorectal carcinoma tissues. Interestingly, in 3 of 5 tumors showing cyclin E gene amplification, the CDK2 gene was amplified simultaneously with rearrangements. No obvious correlation was detected between gene amplification and clinicopathological features of colorectal carcinomas. Out of 7 colon carcinoma cell lines, 2 showed gene amplification of cyclin E without gene amplification of CDK2. No amplification of cyclin A or DI gene was found in any of the colorectal carcinoma tissues or colon carcinoma cell lines. Our results suggest that the concurrent amplification of cyclin E and CDK2 genes may play a role in colorectal carcinogenesis.

Hepatopancreatoduodenectomy for squamous and adenosquamous carcinoma of the gallbladder.

The characteristics of squamous or adenosquamous cell carcinoma of the gallbladder differ quite markedly from those of adenocarcinoma, although the incidence is extremely low. Recently, we encountered both of the former types of gallbladder carcinoma: a 77-year-old man with squamous cell carcinoma and a 70-year-old man with adenosquamous cell carcinoma of the gallbladder. Both had a large mass in the gallbladder fossa region with infiltration to the liver and invasion of the duodenum. Hepatopancreatoduodenectomy was performed on both of these patients. The TNM stage of the former was IV (T4N0M0) and of the latter IV (T4N0M0) and of the latter IV (T4N1bM0). The former has remained well without recurrence for about 1 year and 4 months after the operation, while the latter died of recurrent disease 6 months after operation. The true reason for the difference in the prognosis of these two patients was not known. However, hepatopancreatoduodenectomy is considered to be a most adaptable operative procedure for squamous or adenosquamous cell carcinoma of the gallbladder in view of their mode of spread, and the presence of lymph node metastasis might be a factor of poor prognosis.

Effect of granulocyte colony-stimulating factor (G-CSF) on chemotherapy-induced oral mucositis.

In this study, the ability of granulocyte colony-stimulating factor (G-CSF) to treat or prevent chemotherapy-induced oral mucositis in patients with advanced breast cancer was evaluated. A total of 14 patients who received intraarterial (i.a.) adriamycin (ADM) preoperatively were divided into two groups according to whether or not G-CSF was given. Thus, group A (n = 7) was given G-CSF and group B (n = 7) was not. G-CSF therapy reduced both the incidence and duration of ADM-induced oral mucositis, and a positive correlation was also seen between the incidence of mucositis and ADM-induced leukopenia (< 2,000/mm3). Group A was further divided into two subgroups according to whether G-CSF was given after or before the leukopenia had dropped below 2,000/mm3: group A-1 (n = 3) and group A-2 (n = 4), respectively. ADM-induced mucositis was observed in two of the three patients in group A-1, but in none of the four patients in group A-2. These results strongly support the idea that G-CSF can effectively treat and prevent ADM-induced oral mucositis.

Increased expression and phosphorylation of sh-ptp2 (syp) in human gastric carcinomas.

The expression of SH-PTP2 (SYP), a novel protein tyrosine phosphatase with src homology 2, was examined in human gastric carcinoma cell lines and gastric carcinoma tissues as well as corresponding non-neoplastic mucosas by Northern and Western blotting. The expression of SH-PTP2 mRNA was detected in all 8 gastric carcinoma cell lines at various levels. Most of the cell lines expressed SH-PTP2 protein of 70 kd which corresponds to tyrosine-phosphorylated form, whereas only one cell line expressed unphosphorylated form of SH-PTP2; Fourteen (66.6%) of 21 gastric carcinomas expressed SH-PTP2 mRNA at higher level than non-neoplastic mucosas. Furthermore, most of the gastric carcinomas displayed higher levels of tyrosine-phosphorylated form of SH-PTP2, while non-neoplastic mucosas tended to express unphosphorylated SH-PTP2 protein. These results suggest that the increased expression and tyrosine-phosphorylation of SH-PTP2 may participate in stomach carcinogenesis.

The expression of stat91, ras-independent signal transducer and activator, in human gastric carcinomas.

Upon binding of interferon (IFN) and epidermal growth factor (EGF) to their cell surface receptors, tyrosine phosphorylation of latent cytoplasmic Stat91 (Ras-independent signal transducer and activator of transcription, Stat1 alpha) protein is promptly induced and translocate from the cytoplasm to the nucleus to transduce the signal. The expression of mRNA for Stat91 was examined in 8 gastric carcinoma cell lines and 21 gastric carcinoma tissues as well as corresponding normal mucosa. Of the 8 gastric carcinoma cell lines, all expressed a 4.7 kb Stat91 mRNA and a 91 kD protein at various levels. In gastric carcinoma cell lines, the levels of Stat91 mRNA expression were compatible with those of Stat91 protein expression. In surgical cases, all the gastric carcinoma tissues and their adjacent non-neoplastic mucosa expressed Stat91 mRNA and protein. Interestingly, 14 (66%) out of 21 tumors expressed Stat91 mRNA at higher levels than their corresponding normal mucosas. Moreover, 6 (75%) of 8 tumor tissues expressed higher levels of Stat91 protein as compared with those of the corresponding normal gastric mucosa. No significant correlation was detected between the expression of Stat91 and clinicopathological feature of gastric carcinoma. These results suggest that the majority of gastric cancer in vivo harbour overexpression of Stat91 as a signal transducer in response to various cytokines or growth factors which may be implicated in the growth of gastric cancer.

Interleukin-2 (IL-2) production by human B-cell line.

A human B-cell line (Hairy-BM) constitutively secreting interleukin-2 (IL-2) was established from tumor tissue resected surgically from a patient with breast cancer. Hairy-BM was found to be 100% CD20+, 98% surface immunoglobulin (sIg) G+, 98% sIg kappa chain+, 100% HLA-DR+, 94% IL-2 receptor (IL-2R alpha), 98% IL-2R beta, and devoid of T-cell, monocyte, and natural killer cell surface antigens. The B-cell origin of Hairy-BM was also confirmed by clonally rearranged Ig heavy- and Ig light-chain genes. The growth of Hairy-BM expressing IL-2R was promoted by recombinant IL-2 (rIL-2) and anti-CD25 antibody significantly blocked the growth enhancement. IL-2 secretion from Hairy-BM was confirmed by radioimmunoassay. By using a sensitive polymerase chain reaction technique, we demonstrated that Hairy-BM expressed IL-2 mRNA, IL-2R alpha mRNA, and IL-2R beta mRNA. These findings indicate that certain B-cells not only produce, but also respond to IL-2 in an autocrine fashion with increased proliferation.

A case of hepatocolic fistula after percutaneous drainage for a gas-containing pyogenic liver abscess.

We describe a rare case of gas-containing pyogenic liver abscess which penetrated the adjacent colon, forming a hepatocolic fistula, after percutaneous transhepatic abscess drainage (PTAD) had been performed. To the best of our knowledge, this is the first report of hepatocolic fistula associated with a gas-forming liver abscess in a diabetic patient, with radiological and surgical confirmation of the fistula.

[Hepatic arterial infusion therapy in a patient with multiple hepatic metastases from gastric cancer].

A patient with multiple hepatic metastases (H3) from advanced gastric cancer was treated with pre- and post-operative arterial infusion therapy. A Port-A-Cath was cannulated into the gastroduodenal artery after total gastrectomy. Repeated arterial infusions were performed monthly in the outpatient ward. The volume of hepatic tumors was remarkably decreased to 12% of the pretreatment level. The patient has been alive for 15 months after the operation without any complaint. Arterial infusion chemotherapy through implantable reservoir is useful for the treatment of unresectable hepatic metastasis without decreasing the quality of life.

Increased proliferation of a human breast carcinoma cell line by recombinant interleukin-2.

Two adenocarcinoma cell lines, Breast M25-SF and Breast M, were established from tumor tissue resected surgically from a patient with breast cancer. One, Breast M25-SF, expresses interleukin-2 receptor (IL-2R) on the cell surface and the other, Breast M does not. The effects of recombinant interleukin-2 (rIL-2) on the proliferation of these cell lines were investigated. The growth of Breast M25-SF was significantly promoted by rIL-2 ranging from 1.25 U/ml to 640 U/ml. Anti-CD25 (Tac) antibody significantly blocked the growth enhancement of Breast M25-SF by rIL-2. Breast M, however, did not respond to rIL-2. To confirm more directly the promotion of Breast M25-SF growth by rIL-2, cloning of IL-2 responders from parent Breast M25-SF cells was carried out by limiting dilution without feeder cells in 96-well microplates. No colony formation was found in 24 wells without rIL-2. Eleven, 13 and 6 clones were established from groups of 24 wells containing rIL-2 at 200, 20 and 2 U/ml respectively. All of the clones expressed IL-2R and respond to rIL-2. By using a sensitive polymerase chain reaction technique, we demonstrated that Breast M25-SF but not Breast M expressed IL-2 mRNA, and IL-2 secretion from Breast M25-SF but not Breast M was also confirmed by radioimmunoassay. These findings suggest a role for IL-2 in autocrine support of Breast M25-SF growth. IL-2 may play an important role in the growth control of breast carcinoma cells.

Multicentre randomised study of adjuvant chemotherapy with mitomycin C and tegafur or tegafur-uracil in gastric cancer.

OBJECTIVE: To compare therapeutic results and five year survival after two regimens of adjuvant chemotherapy after resections for gastric cancer. DESIGN: Prospective randomised multicentre trial. SETTINGS: 21 departments of surgery. SUBJECTS: 243 patients with stage II, III, or IV gastric cancer. INTERVENTIONS: All patients received an intravenous bolus of mitomycin C 20 mg on the day of operation, and 10 mg on the first postoperative day. They were then randomised to receive either tegafur 600 mg or tegafur-uracil 600 mg orally daily beginning two weeks after operation and continuing for two years. Patients with histological confirmed stage IV disease also received mitomycin C 10 mg every three months starting one month after operation. MAIN OUTCOME MEASURES: Survival and side effects. RESULTS: 13 patients (5%) were withdrawn from the study, leaving 230 for analysis. There were no serious side effects and both drugs were safe when given continuously for two years. There were no significant differences in 5 year survival (though patients who were given tegafur-uracil tended to live longer), except when patients who had histological curative resections were compared with and those with poorly differentiated adenocarcinoma (p = 0.04).

An unusual presentation of spontaneous pneumoperitoneum secondary to the rupture of a gas-containing pyogenic liver abscess: report of a case.

We describe a rare case of spontaneous pneumoperitoneum secondary to the rupture of a gas-containing pyogenic liver abscess in a 59-year-old man. The patient was diagnosed as having a hollow viscus perforation based on a sudden onset of acute abdominal pain along with radiological evidence of bilateral subphrenic feee air (pneumoperitoneum), and underwent an emergency laparotomy. Contrary to expectations, the surgery revealed no perforations of the hollow viscus, but instead a ruptured liver abscess at the dome of the right hepatic lobe was identified associated with suppurative peritonitis. To the best of our knowledge, such a case of spontaneous pneumoperitoneum secondary to the rupture of a gas-containing liver abscess is extremely rare.

Inhibition of tumor cell growth by a human B-cell line.

An Adenocarcinoma cell line (Breast-M) and an Epstein-Barr virus (EBV)-infected B-cell line (Hairy-BM) were established from breast tumor tissue. The Hairy-BM was CD20+, CD25 (Tac)+ and surface immunoglobulin (sIg)+. Hairy-BM suppressed the in vitro proliferation of Breast-M in a time- and a dose-dependent manner. The suppression was also found in 5 different human tumor targets showing tumor-Hairy-BM binding, but not; in 2 murine tumor targets showing no significant tumor-Hairy-BM binding. Lytic activity of Hairy-BM was found only against Breast-M.

Differentially potentiating effects by dipyridamole on cytotoxicity of 5-fluorouracil against three human maxillary cancer cell lines derived from a single tumor.

Dipyridamole (DPM), an inhibitor of nucleoside transport, is a unique potentiator of 5-fluorouracil (5-FU). We examined the combined effects of DPM and 5-FU on three cell lines (IMC-2, IMC-3, IMC-4) derived from a single tumor of a patient with maxillary cancer. DPM at 1.0-2.0 microM potentiated the cytocidal action of 5-FU > 20-fold against IMC-2 cells, while DPM at 10-20 microM potentiated the action of 5-FU against IMC-3 cells by 2-fold and against IMC-4 cells by approximately 4-fold. We examined why DPM differentially potentiated 5-FU against the three cell lines. The three maxillary cell lines showed heterogeneous sensitivities to the combination of 5-FU and DPM. We compared intracellular metabolism of 5-FU in IMC-2, IMC-3 and IMC-4 cells by HPLC. Determination of 5-FU metabolites demonstrated that cellular contents of 5-fluorodeoxyuridine monophosphate (FdUMP) were 104.66, 15.35 and 61.5 fmol/10(6) cells in IMC-2, IMC-3 and IMC-4 cells, respectively, in the presence of 10 microM DPM when the three cell lines had similar levels (0.21-0.33 fmol/10(6) cells) of FdUMP in the absence of DPM. By contrast, cellular levels of 5-FU and 5-fluorouridine triphosphate (FUTP) were not appreciably changed by DPM in three cell lines. The cellular level of thymidylate synthase mRNA in IMC-2 cells was found to be about one-fifth of that in IMC-3 and IMC-4 cells. The differential effects of DPM on the potentiation of 5-FU cytotoxicity might be closely related to the cellular levels of FdUMP and its target enzyme, thymidylate synthase in three human maxillary cancer cell lines.