Noncycling tumor cells: mitogenic response to antilymphocytic serum.

After continuous labeling with tritiated thymidine for a period several times the cell generation time, some ELD ascites cells remained unlabeled. Despite continued exposure to tritiated thymidine, unlabeled mitoses appeared promptly after administration of mouse antilymphocytic serum. Immunosuppression released some noncycling G(2) tumor cells into mitosis.

Breast cancer: induction of differentiation by embryonic tissue.

A mouse mammary tumor, adenocarcinoma BW 10232, was maintained in vitro for 14 days, separated from embryonic mammary mesenchyme by a Millipore filter. Tubules developed in the tumor; deoxyibonucleic acid synthisis declined; and a presumptive acid mucopolysaccharide matrix, not evident in the controls, appeared.

Effect of wheat fiber and vitamins C and E on rectal polyps in patients with familial adenomatous polyposis.

Over a 4-year period in a chemoprevention trial on large bowel neoplasia, 58 patients with familial adenomatous polyposis were treated with 4 g of ascorbic acid (vitamin C)/day plus 400 mg of alpha-tocopherol (vitamin E)/day alone or with a grain fiber supplement (22.5 g/day). In this randomized, double-blind, placebo-controlled study, we determined the effects of these supplements on rectal polyps in these patients. Analysis by intent to treat suggested that the high-fiber supplement had a limited effect. Analysis adjusted for patient compliance showed a stronger benefit from the high-fiber supplement during the middle 2 years of the trial. The results provide evidence for inhibition of benign large bowel neoplasia by grain fiber supplements in excess of 11 g/day in this study population. The findings are consistent with the hypothesis that dietary grain fiber and total dietary fat act as competing variables in the genesis of large bowel neoplasia.

Embryonic inductive tissues that cause histologic differentiation of murine mammary carcinoma in vitro.

A murine mammary tumor was cultured in vitro for 14 days, either in direct combination with various embryonic murine inductive tissues or separated by a Millipore filter from these tissues. From 456 test cultures and 269 control cultures of tumor alone, morphologic, histochemical, and autoradiographic evidence for cytodifferentiation was obtained in the tumor after exposure to inductive tissues directly or through the filter. There appeared to be a gradient in potency of the inductive tissues; embryonic mammary mesenchyme was the most active of the tissues tested. Tumor growth was not different from that of controls, however, when the cultured, inductive tissue-exposed neoplasm was returned to the murine host.

Acute effects of dietary cholic acid and methylazoxymethanol acetate on colon epithelial cell proliferation; metabolism of bile salts and neutral sterols in conventional and germfree SD rats.

The acute effects of cholic acid ingestion on methylazoxymethanol acetate [(MAM) CAS: 592-62-1]-treated conventional and germfree rats were investigated. Male SD rats were divided into 4 treatment groups. The first group received control chow; the second group, control chow plus 0.5% cholic acid; the third group, control chow plus MAM; and the fourth group, control chow plus 0.5% cholic acid plus MAM. Fecal bile acids, cholesterol, cholesteral degradation products, and neutral sterols, as well as labeling indices and numbers of epithelial cells per crypt column, were measured after 6 weeks of treatment. The administration of MAM to germfree groups diminished both fecal bulk and the amount of fecal water. MAM did not affect the fecal bile acid composition. Analysis of the fecal bile acids in conventional rats fed cholic acid demonstrated that half of the bile acids were in a form of deoxycholic acid. In the germfree groups fed cholic acid, 90% of the bile acids appeared unaltered in the feces. Neither in the germfree nor in the conventional groups was an effect seen of MAM on the output of fecal neutral sterols. The addition of cholic acid to the food decreased the output of neutral sterols both in the conventional (P less than .001) and in the germfree (P less than .02) animals. The germfree animals showed a reduced amount of neutral steroid excretion (P less than .01) when compared to the findings for the conventional groups. MAM had no influence on the fecal cholesterol or coprostanol output. The consumption of 0.5% cholic acid decreased the total output of cholesterol (P less than .05). The excretion of coprostanol was significantly diminished in the conventional rats fed cholic acid (P less than .001). No difference in labeling indices was observed between conventional and germfree rats, whether treated with cholic acid, MAM, or cholic acid plus MAM. However, all germfree groups showed less epithelial cells per crypt column (P less than .001) than did conventional animals.

Effect of flurbiprofen and 16,16-dimethyl-prostaglandin E2 on gastrointestinal tumorigenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in rats. I. Squamous epithelium and mesenchymal tissue.

The effect of exogenous synthetic prostaglandins and the inhibition of endogenous prostaglandin synthesis on gastrointestinal tumorigenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine [(MNNG) CAS: 70-25-7] was studied in female Wistar rats (100 g). Animals were divided into 6 groups: Group I was treated with MNNG alone (No. = 43); group II was treated with MNNG after application of the cyclo-oxygenase inhibitor flurbiprofen (No. = 44); group III was treated with MNNG after oral administration of 16,16-dimethyl-prostaglandin E2 (16,16-dm-PGE2; No. = 43); group IV received flurbiprofen alone (No. = 15); group V was treated with 16,16-dm-PGE2 alone (No. = 11). Animals in group VI served as controls (No. = 15). All drugs were administered orally. Hyperkeratosis and hyperplasia of the forestomach developed by 10 days after the first treatment with the carcinogen. Later, benign papillomas and dysplastic lesions were noted. Progressive ingrowth of squamous epithelium from the forestomach ridge into the glandular stomach started as early as day 13 and was more frequent in animals treated with a combination of MNNG plus flurbiprofen (P less than .001). The first squamous cell carcinomas of the forestomach were detected on day 51. Their incidence was 38, 60, and 42% in groups I, II, and III, respectively. This difference was not statistically significant. The incidence of mesenchymal tumors (leiomyosarcoma) in the stomach and duodenum was higher following treatment with MNNG plus flurbiprofen as compared to the incidence following treatment with MNNG alone or in combination with 16,16-dm-PGE2 (P less than .005). No malignant tumors developed in the gastrointestinal tracts of animals treated with flurbiprofen or 16,16-dm-PGE2 alone or in controls. The higher incidence of gastric and duodenal leiomyosarcomas after treatment with flurbiprofen suggests that reduction of prostaglandin synthesis favored the development of MNNG-induced cancer in mesenchymal tissues of the upper gastrointestinal tract.

Effect of flurbiprofen and 16,16-dimethyl prostaglandin E2 on gastrointestinal tumorigenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in rats: glandular epithelium of stomach and duodenum.

The effect of an exogenous synthetic prostaglandin analogue, 16,16-dimethyl prostaglandin E2 (16,16-dm-PGE2), as well as the effect of endogenous prostaglandin synthesis inhibition by a cyclooxygenase inhibitor, flurbiprofen, on chemically induced gastric carcinogenesis has been investigated in rats. Carcinogenesis was induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; CAS:70-25-7). Animals were divided into six groups: Group I, treatment with MNNG alone; Group II, treatment with 16,16-dm-PGE2 plus MNNG; Group III, treatment with flurbiprofen plus MNNG; Group IV, treatment with 16,16-dm-PGE2 alone; Group V, treatment with flurbiprofen alone; and Group VI, controls. Treatment with high doses of MNNG resulted in rapid development of malignant tumors originating from the glandular epithelium of the stomach and duodenum in animals of all groups receiving the carcinogen. The first gastric adenocarcinoma infiltrating the muscularis proper was detected after 139 days in an animal treated with a combination of MNNG and flurbiprofen. The incidence of infiltrating adenocarcinoma and the incidence of all neoplastic lesions of the glandular stomach were both significantly higher in animals treated with a combination of MNNG and flurbiprofen compared with treatment by MNNG alone or in combination with 16,16-dm-PGE2 (P less than 0.05 and P less than 0.001). The difference in tumor incidence between the last two groups was not significant. The first duodenal adenocarcinoma was detected on Day 114 in another animal of the group treated with MNNG plus flurbiprofen. When compared with the group treated with MNNG plus 16,16-dm-PGE2, significantly more animals developed duodenal adenocarcinoma when treated with MNNG plus flurbiprofen (P less than 0.005) or with MNNG alone (P less than 0.05). Results of this study indicate that inhibition of endogenous prostaglandin synthesis favors development of adenocarcinoma in the glandular stomach of rats. Vice versa, the addition of an exogenous prostaglandin analogue inhibits the development of duodenal adenocarcinoma. This protective effect of prostaglandins may be due to an increase of the thickness of the mucus gel covering the glandular epithelium, thereby preventing access of carcinogen to the mucosa.

Sodium transport in a mouse model of colonic carcinogenesis.

Following 4 weeks of s.c. injections of 1,2-dimethylhydrazine, a carcinogen that produces colon cancer in CF1 mice, an increase in the unidirectional mucosal to serosal flux and net absorption of sodium was observed in the distal colon. This increase in sodium transport was amiloride sensitive. 1,2-Dimethylhydrazine treatment had no effect on sodium transport in the distal colon of DBA/2 mice, a strain which does not develop colonic malignant transformation. Although stimulation of sodium transport has been observed in cultured cell systems exposed to growth factors, similar changes in sodium transport have not previously been demonstrated in an intact epithelium at an early stage of carcinogenesis. The present study in mouse distal colon demonstrates that sodium transport is altered in 1,2-dimethylhydrazine-induced malignant transformation of the large bowel.

Loss of colonic HLA antigens in familial adenomatous polyposis.

The loss of HLA antigens by neoplastic cells is considered important for tumor growth and metastasis, since it may allow tumors to escape immune surveillance. We studied the expression of HLA class I and II antigens in the colons of 10 patients with familial adenomatous polyposis (FAP), a condition which leads inevitably to colorectal cancer. Expression of HLA class antigens was studied by immunohistochemistry in (a) adenomas from patients with FAP, (b) histologically normal mucosa distant from the adenomas, and (c) histologically normal colonic mucosa from normal subjects. The expression of HLA class I and II antigens was decreased in histologically normal mucosa from FAP patients compared to normal controls. Adenomas showed a similar but quantitatively more pronounced reduction (or loss) of HLA antigen expression. The reduction of HLA expression in adenomas was comparable to that observed in sporadic colon carcinomas. This generalized suppression of HLA gene expression in the colon of FAP patients, which precedes the onset of overt histological manifestations of neoplasia, may be an important early event in colon carcinogenesis.

Reduced expression of HLA class I and II antigens in colon cancer.

The expression of HLA class I and II antigens was studied by immunohistochemistry in (a) specimens of colon cancer from 25 patients, (b) normal colonic mucosa obtained 5-10 cm away from each tumor, and (c) colonic mucosa from 13 normal individuals. Thirteen of the tumor specimens had normal epithelium adjacent to the cancer, which thus served as an internal control. The expression of HLA class I antigens in colon cancer was dramatically reduced compared to control (P less than 0.0001): undetectable in 28%, diminished in 68%, normal in 4%. The expression of class II antigens was also reduced in cancer (P less than 0.0001 for all when compared to normal), being undetectable in most (HLA-DP 64%, HLA-DQ 72%, HLA-DR 68%). In 44% of the cancers all three HLA class II antigens were undetectable; in 92% at least one class II antigen was undetectable; and in 20% both class I and class II antigens were undetectable. No cancer specimen had a completely normal HLA phenotype. The expression of other surface antigens was preserved in cancer tissues and, therefore, loss of HLA antigens was not due to a nonspecific decline in surface molecules. When glands of normal mucosa immediately adjacent to cancer were compared to those of normal controls, significantly reduced expression of only HLA class I antigens (P = 0.0149) and HLA-DP (P = 0.034) was found. The expression of the HLA antigens in colonic mucosa remote from the cancer was no different from that of normal controls. Our data show extensive and significant reduction in the expression of HLA antigens in colon cancer; its potential relationship to immunosurveillance in cancer is discussed.

Cyclooxygenase-2 overexpression and tumor formation are blocked by sulindac in a murine model of familial adenomatous polyposis.

Inducible cyclooxygenase (Cox-2), also known as prostaglandin H synthase 2 (PGH-2) is a key enzyme in the formation of prostaglandins and thromboxanes. Cox-2 is the product of an immediate-early gene that is expressed in response to growth factors, tumor promoters, or cytokines. Overexpression of Cox-2 is associated with both human colon cancers and suppression of apoptosis in cultured epithelia] cells, an activity that is reversed by the nonsteroidal anti-inflammatory drug, sulindac sulfide. To address the relationship between Cox-2, apoptosis, and tumor development in vivo, we studied C57BL/6J-Min/+(Min) mice, a strain containing a fully penetrant dominant mutation in the Apc gene, leading to the development of gastrointestinal adenomas by 110 days of age. Min mice were fed AIN-76A chow diet and given sulindac (0.5 +/- 0.1 mg/day) in drinking water. Control Min mice and homozygous C57BL/6J-+/+ normal littermates lacking the Apc mutation (+/+) were fed AIN-76A diet and given tap water to drink. At 110 days of age, all mice were sacrificed, and their intestinal tracts were examined. Control Min mice had 11.9 +/- 7.8 tumors per mouse compared to 0.1 +/- 0.1 tumors for sulindac-treated Min mice. As expected, +/+ littermates had no macroscopic tumors. Examination of histologically normal-appearing small bowel from Min animals revealed increased amounts of Cox-2 and prostaglandin E(2) compared to +/+ littermates. Using two different in situ techniques, terminal transferase-mediated dUTP nick end labeling and a direct immunoperoxidase method, Min animals also demonstrated a 27-47% decrease in enterocyte apoptosis compared to +/+ animals. Treatment with sulindac not only inhibited tumor formation but decreased small bowel Cox-2 and prostaglandin E(2) to baseline and restored normal levels of apoptosis. These data suggest that overexpression of Cox-2 is associated with tumorigenesis in the gastrointestinal epithelium, and that both are inhibited by sulindac administration.

Orchiectomy alone in the treatment of clinical stage I nonseminomatous germ cell tumor of the testis.

Forty-five patients with clinical stage I nonseminomatous germ cell tumor of the testis (NSGCTT) were entered in a prospective clinical trial to receive no treatment other than orchiectomy until clinical evidence of relapse. Of this group, 36 patients (80%) have been continuously free of disease for a median duration of 19.5 months after orchiectomy. Nine patients (20%) have relapsed, eight within seven months of orchiectomy. Seven of nine relapsing patients have been rendered free of disease with chemotherapy and/or surgery for a median duration of seven months (range, one to 33 months) after completion of treatment; the other two patients are presently under treatment although one has progressive disease. The relapse rate was higher in patients with embryonal carcinoma than in those with teratocarcinoma, 57% versus 17%. These preliminary results imply that the omission of routine lymphadenectomy or lymph-node irradiation in clinical stage I NSGCTT deserves further trial.

Surgical prophylaxis of familial colon cancer: prevention of death from familial colorectal cancer.

Patients at risk for inherited colorectal cancer constitute a heterogeneous population. A total colectomy is minimal treatment for those patients with invasive cancer or those with established risk factors. For others at risk, predictive genetic markers, correlated with clinical and pathologic determinants, will establish the basis for policies of surveillance and preventive surgery.

Primary lymphomas of the gastrointestinal tract. Analysis of prognostic factors with emphasis on histological type.

We have analyzed the clinicopathological factors affecting survival in 60 primary gastrointestinal lymphomas seen at Memorial Hospital between 1949 and 1978. Patients with generalized lymphoma (Stages III and IV) at the time of diagnosis and those without follow-up information or adequate histological material were excluded from this study. Lymphomas were classified according to the Lukes-Collins, Kiel, and Rappaport schemes and the patients were staged retrospectively by a modified Ann Arbor system. The patients were treated by surgical resection, radiotherapy, or both. Survival was influenced by histological type (P = 0.0116), stage of the disease (P less than 0.0001), and size of the primary tumor (P = 0.0007). Low-grade lymphoplasmacytoid lymphomas, recognized in 26.6% of the cases, had a low rate of extra-abdominal recurrence; 74% of these patients were alive without evidence of recurrence after a median follow-up of 171 months, or died without evidence of lymphoma with a median survival of 147 months. Centrocytic (Kiel) or cleaved cell (Lukes-Collins) types were seen in 13% and 21%, and high grade (Kiel) or large noncleaved and immunoblastic (Lukes-Collins) in 33.3% and 30% of the cases, respectively. These groups had a high rate of extra-abdominal recurrences, and over 60% of the patients died of lymphoma, with a median survival of 8 for the centroblastic-centrocytic and 7 months for the high-grade tumors. Histological type and clinicopathological staging emerge as useful factors for the identification of patients with high risk of systemic recurrence, probably best treated with chemotherapy in addition to surgery and local radiotherapy.

A morphologic and kinetic basis for the more invasive character of N-methyl-N'-nitro-N-nitrosoguanidine induced duodenal tumors following pyloroplasty.

Pyloroplasty increased the invasiveness of duodenal tumors in Wistar rats receiving N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) for 12 weeks (83 mg/l in drinking water) (Arch. Surg., 117, 768-771). To assess morphologic and kinetic alterations, analyses of tritiated thymidine (3HTdR) pulse-labeled fundic, antral and duodenal mucosa were carried out. The normal appearance of MNNG-treated antral and duodenal mucosa was characterized by the appearance of elongated hyperactive columns exhibiting elevated levels of DNA synthesis. Pyloroplasty in carcinogen-treated rats induced both a 3-fold enhancement in the number of these elongated columns and an elevation in the number of proliferating cells within them (P less than 0.001). In the MNNG and pyloroplasty treated duodenal mucosa 26% of columns contained over 130 cells/column rather than an average of 100 cells in normal appearing MNNG-treated mucosa. DNA synthesis was increased by 23% within these hyperactive glands (11.3 proliferative cells/column vs. 9.4/column in normal appearing mucosa). Pyloroplasty creates both an increase of gastric bile reflux and an increase of the gastric evacuation rate, conditions which influence cell proliferation. Such alterations in antro-pyloro-duodenal physiology contribute to the increased cellular activity observed, promote malignant transformation and foster the expression of invasiveness.

Hyperpigmented lesions of the retinal pigment epithelium in familial adenomatous polyposis.

Ophthalmic examinations were performed on 56 patients with validated familial adenomatous polyposis (FAP) for hyperpigmented defects of the retinal pigment epithelium. Such lesions were seen bilaterally in 29 patients (52%) and unilaterally in 8 patients (14%). Of the 56 patients, 33 had one or more of the extracolonic expressions associated with Gardner syndrome. We found retinal lesions in 8 patients without any of the expressions of Gardner syndrome. No association was found between Gardner syndrome and the retinal lesions when these patients were compared to patients without any stigmata of Gardner syndrome, nor was any significant association found when each of the expressions was compared individually with the presence of the pigmented retinal lesions. The presence or absence of eye findings were seen to cluster within families. There was no association with sex. Fundus lesions are apparently a variable expression of the FAP gene and are not specifically associated with Gardner syndrome.

Evidence for adenoma-carcinoma sequence in the duodenum of patients with familial adenomatous polyposis. The Leeds Castle Polyposis Group (Upper Gastrointestinal Committee).

AIMS: To explore the association between duodenal adenoma and carcinoma in patients with familial adenomatous polyposis (FAP). METHODS: A multicentre survey of 1262 patients with FAP yielded 47 cases of duodenal cancer. The association between adenoma and cancer was assessed in these cases. RESULTS: Adenomatous tissue was found within duodenal cancer in 29 of 44 (66%) patients with FAP and in mucosa adjacent to duodenal cancer in 31 of 42 (73%) such patients. Adenomas were found as a component of, or adjacent to, duodenal cancer in 38 of 45 (84%) patients. CONCLUSIONS: These observations support the existence of the adenomacarcinoma sequence in the duodenum of patients with FAP. Factors associated with malignant change included villous histology, moderate or severe dysplasia, and the presence of stage IV duodenal polyposis.

Sphincter-sparing surgery.

Since World War II, a variety of technical innovations have been introduced to preserve the anal sphincter in patients with chronic ulcerative colitis or tumors of the rectum. Studies of anorectal physiology have yielded guidelines for preserving continence and minimizing morbidity. For example, preservation of the anal rectal angle is essential; the rectal mucosa can be removed without impairing neural mechanisms; construction of a reservoir increases rectal compliance. Following these guidelines, surgeons have introduced new techniques and applications benefiting a larger and more varied patient population. Some technical problems remain, however, and patient selection criteria for restorative rectal surgery need to be refined.

Promotion of gastric tumorigenesis by duodenal contents in rats induced with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG).

Reflux of duodenal contents into the stomach occurs in patients with pyloric incompetence and after gastric resection when bile-diverting procedures are omitted. In such settings duodenal contents have been considered to favor the development of gastric cancer. We have studied the effect of chronic duodenogastric reflux on gastric tumor promotion in rats treated with N-methyl-N'-nitrosoguanidine (MNNG) in an experimental design that avoids physical trauma to the glandular stomach. Thus the effect of trauma-induced tissue repair on carcinogenesis is eliminated, and duodenogastric reflux is isolated as an experimental parameter. To achieve such reflux the first jejunal loop was anastomosed to the forestomach in rats. Animals were exposed to MNNG in drinking water (83 mg/L) for 12 weeks before induction of reflux. Experimental groups were as follow: I, reflux plus MNNG (n = 32); II, MNNG alone (n = 27); III, reflux alone (n = 28); IV, control (n = 25). The experiment was terminated after 56 weeks. Only animals that had survived for 90 days were included in the effective number of animals, which allowed for equal chances of tumor development. In no animal that died earlier had tumors developed. Animals with reflux plus MNNG treatment had significantly more glandular neoplasms (12/32) than did animals with MNNG treatment alone (4/27; p less than 0.05). Similarly, more animals with squamous cell neoplasms were recorded in group I (9/32) than in group II (2/27; p less than 0.05). In consideration of all tumors of epithelial and mesenchymal origin, more gastric malignant tumors were observed in group I (9/32) than in group II (2/27; p less than 0.05). It is concluded that chronic exposure to duodenal contents promotes the development of gastric neoplasia.

Intestinal motility changes in experimental large bowel obstruction.

Bipolar electrodes for recording electrical discharges of colon smooth muscle and strain gages for recording associated contractions of circular muscle were implanted in six rhesus monkeys. After recovery, baseline records were made. The animals then had an obstruction device implanted in sigmoid colon which resulted in progressive compromise of the lumen terminating in complete obstruction after 13 +/- 2 days. Recording were made daily during development of obstruction. As obstruction became more complete, contraction frequency decreased in right colon, increased in left colon proximal to the obstruction, and was unchanged in left colon distal to the obstruction. The frequency of distentions increased in colon proximal to the obstruction but was unchanged distally. Simultaneous mass actions, a complex of nonperistaltic high amplitude contractions and distentions occurring nearly simultaneously throughout the colon which is not seen in normal colon, appeared in colon both proximal and distal to the obstruction and became more frequent as the degree of obstruction progressed. Colon obstruction results in abnormal motility complexes, but not in hyperperistalsis. Mass actions probably are the basis for colic and rushing bowel sounds noted clinically in colon obstruction.