Sustained viral response to pegylated interferon alpha-2b and ribavirin in chronic hepatitis C refractory to prior treatment.

Hepatitis C virus (HCV) infection refractory to previous therapy is common. Treatment of patients with refractory disease is difficult and less studied. Pegylated interferon alpha-2b plus ribavirin is used for treatment of HCV patients naïve to therapy. We conducted a randomized study for refractory HCV patients using a high- vs. a low-dose pegylated interferon alpha-2b and ribavirin protocol. Our aim was (1) to determine the efficacy of pegylated interferon alpha-2b plus ribavirin to eradicate HCV in previously treated individuals and (2) to compare a low-dose to a high-dose regimen. One hundred fifty-two patients were initiated in the study, 112 (74%) were male and 40 (26%) female. Nineteen percent of patients obtained a sustained viral response (SVR) in the high-dose arm. Prior relapsers had the highest SVR rates: 50% in non-genotype 1 and 34% in genotype 1. The odds of achieving a SVR were six times higher in previous relapsers. The rate of SVR in genotype 1 patients who were nonresponders to prior therapy was only 8%. All patients who achieved a SVR had no detectable virus at week 24. However, only half of those who had undetectable viral titers at week 24 achieved a SVR. In conclusion, retreatment of patients with refractory hepatitis C infection with interferon alpha-2b and ribavirin combination therapy is well tolerated and gives modest response rates. The most important factor in predicting response to therapy is the manner of response to previous treatment. The likelihood of response to treatment can be predicted from the viral titers at 24 weeks.

Apoptosis associated with esophageal adenocarcinoma: influence of photodynamic therapy.

Tumor cell death in vitro by photodynamic therapy (PDT) has been related to the induction of apoptosis. We measured and compared changes in apoptosis and caspase 3 activity, an effector of apoptosis, in normal and neoplastic esophageal tissues during PDT. Apoptosis index, caspase 3 cleavage activity, pro-caspase 3, p53, and bcl-2 levels were measured in normal and neoplastic tissues of patients with esophageal adenocarcinoma before, during, and after PDT with Photofrin. The apoptotic index was greater in carcinoma tissue compared to adjacent normal tissues. In concert, pro-caspase 3 immunoreactivity was absent and caspase 3-like cleavage activity was over 30-fold greater in carcinoma tissue compared to normal esophageal tissues. These parameters were unaffected by PDT. Variable changes in bcl-2 and p53 immunoreactivity were noted in normal and carcinoma tissues during PDT. Greater levels of apoptosis and caspase 3 activity are hallmarks of esophageal adenocarcinoma compared to normal esophageal tissue. These differences were unaffected by PDT. This may be due to the fact that tissues were obtained 72 h post-PDT therapy. Changes in these parameters may have occurred early after PDT therapy. An assessment of apoptosis and caspase 3 activity prior to 72 h post-PDT may provide further insight into the mechanism involved, although no sustained effects on these parameters by PDT were noted.

Overexpression of epidermal growth factor receptor in Peutz-Jeghers syndrome.

Peutz-Jeghers syndrome is characterized by gastrointestinal hamartomatous polyposis, mucocutaneous pigmentation, and a predisposition to cancer. The etiology of this syndrome is unknown. We investigated the expression of epidermal growth factor receptor (EGFr), transforming growth factor-alpha (TGF-alpha), transforming growth factor-beta1 (TGF-beta1) and transforming growth factor-beta receptor (TGF-beta RII) between normal and Peutz-Jeghers small bowel tissues. In addition, immunoprecipitation by phosphotyrosine antibodies followed by EGFr western blotting was measured and compared between a Peutz-Jeghers hamartoma and normal duodenal tissue. EGFr expression was increased 2.5-fold in normal and hamartomatous tissue of Peutz-Jeghers patients compared to normal small bowel tissue. In Peutz-Jeghers tissues, the major EGFr immunoreactive band was increased size from 170 to approximately 200 kDa. Using an antibody specific for activated EGFr, this larger size band was predominant in Peutz-Jeghers tissue. Immunoprecipitation of a hamartoma by a phosphotyrosine specific antibody followed by western blotting for EGFr demonstrated this 200-kDa band. Expression of TGF-alpha, TGF-beta1, TGF-beta1 RII was not significantly different between normal and Peutz-Jeghers tissues. In conclusion, EGFr was overexpressed in normal and hamartomatous small bowel tissue of Peutz-Jeghers patients, which suggests that EGFr in Peutz-Jeghers tissue is persistently activated or highly stimulated by endogenous ligands and also suggests a possible role for EGFr in the pathogenesis of Peutz-Jeghers syndrome.

Differential effects of deoxycholic acid on proliferation of neoplastic and differentiated colonocytes in vitro.

The secondary bile acid deoxycholic acid is believed to be a promoter of large bowel cancer, in part by inducing colonic epithelial proliferation. The effects of deoxycholic acid on [3H]thymidine incorporation by the human colon cancer cell line HT29 and two differentiated subclones were measured and compared. The subclone HT29-C1 has features of mature absorptive cells and HT29-N2 cells secrete mucus under cholinergic control. The three cell lines were treated with deoxycholic acid (DCA) at concentrations of 0, 5, 10, 50, 100, 150, and 300 microM for 3, 6, 9, 15, 24, and 48 hr. A significant increase in proliferation was noted in HT29 cells only at 6 hr with 5 and 10 microM deoxycholic acid. Neither the subclone HT29-C1, nor HT29-N2 cells exhibited significant change in [3H]thymidine incorporation with DCA at these concentrations or time points. Higher doses of deoxycholic acid above 50 microM and duration of exposure greater than 24 hr were cytotoxic to all three cell lines. The proliferative effects of DCA in HT29 cells were not paralleled by changes in protein kinase C activity or protein kinase C isoform expression. Quantitative and qualitative differences in PKC isoform expression were not noted in the three cell lines used in this study. The proliferative effects of DCA on HT29 cells appear to be independent of the PKC signal transduction pathway.

The epidermal growth factor receptor (EGF-R) is present on the basolateral, but not the apical, surface of enterocytes in the human gastrointestinal tract.

BACKGROUND: While it is clear that luminal epidermal growth factor (EGF) stimulates repair of the damaged bowel, its significance in maintaining normal gut growth remains uncertain. If EGF is important in maintaining normal gut growth, the EGF receptor (EGF-R) should be present on the apical (luminal) surface in addition to the basolateral surface. AIMS/SUBJECTS/METHODS: This study examined the distribution of the EGF-R in the epithelium throughout the human gastro-intestinal tract using immunohistochemistry, electron microscopy, and western blotting of brush border preparations. RESULTS: Immunostaining of the oesophagus showed circumferential EGF-R positivity in the cells of the basal portions of the stratified squamous epithelium but surface cells were EGF-R negative. In the normal stomach, small intestine, and colon, immunostaining localised the receptor to the basolateral surface with the apical membranes being consistently negative. EGF-R positivity within the small intestine appeared to be almost entirely restricted to the proliferative (crypt) region. Western blotting demonstrated a 170 kDa protein in whole tissue homogenates but not in the brush border vesicle preparations. CONCLUSIONS: As the EGF-R is located only on the basolateral surfaces in the normal adult gastrointestinal tract, the major role of luminal EGF is probably to stimulate repair rather than to maintain normal gut growth.

Localization of protein kinase C alpha isoform expression in the human gastrointestinal tract.

Protein kinase C (PKC) includes a family of related proteins which constitutes a major signal transduction pathway. The aim of this study was to determine the localization of the PKC-alpha isoform throughout the human gastrointestinal tract. PKC-alpha expression was also measured and compared between normal and neoplastic colorectal tissue. PKC-alpha mRNA expression was detected in normal human gastrointestinal tract tissue using Northern blot analyses and in situ hybridization. PKC-alpha protein expression was detected in normal gastrointestinal tissue and colorectal neoplasia using Western blot and immunohistochemical analyses. PKC-alpha was expressed throughout the human gastrointestinal tract. Distinct organ and cellular localization was characterized. PKC-alpha mRNA and protein localization were most prevalent in the deep basal layer of the esophageal mucosa. In the stomach, PKC-alpha expression was detected predominately in the cells of the deep glands and surface epithelial cells but less in the mucous neck cells of the gastric pits. In the duodenum and ileum, PKC-alpha mRNA expression was greater in the deeper crypt cells than in the differentiated cells that line the villi. However, immunohistochemistry showed greater expression in the cells of the villi compared to crypt cells. In normal colonic tissue, PKC-alpha mRNA and protein predominated in the cells of the upper crypt and surface epithelial cells. PKC-alpha protein was also prominently expressed in the glands of colorectal adenocarcinoma. There was no quantitative difference in the level of PKC-alpha protein expression between normal and neoplastic colorectal tissue. The specific organ and cellular expression of PKC-alpha suggests separate and distinct functional roles for this PKC isoform throughout the gastrointestinal tissues.

Protein kinase C activity as a potential marker for colorectal neoplasia.

Protein kinase C (PKC) is a mediator of transmembrane signal transduction, important in cell growth and differentiation. Cell activation by extracellular signals is associated with a translocation of PKC from the cytosol to the membrane. We measured and compared PKC activity in cytosol and membrane fractions of normal and neoplastic colorectal tissue. Total and membrane-associated PKC activity in normal colorectal tissue was greater in patients (N = 16) with colorectal cancer compared to that from patients with a normal colonoscopy (N = 16), P < 0.01. A similar trend was noted in PKC activity of normal colorectal tissue from patients with adenomas compared to patients with a normal colonoscopy. PKC activity (total, membrane-associated, percent membrane) was not different in neoplastic colorectal tissue compared to that of adjacent normal tissue. However, there was a considerable range of PKC activity noted in all groups, which would limit the utility of PKC activity as a marker for colorectal neoplasia.

Selenium and difluoromethylornithine additively inhibit DMH-induced distal colon tumor formation in rats fed a fiber-free diet.

We investigated the effects of difluoromethylornithine, an inhibitor of ornithine decarboxylase (ODC) and selenium supplementation on tumor formation induced by the carcinogen 1,2-dimethylhydrazine (DMH) in Sprague-Dawley rats. A biochemical link between polyamine biosynthesis and selenium metabolism to its cancer preventative form has been suggested by the common requirement of S-adenosylmethionine. One-hundred and twenty male Sprague-Dawley rats were divided into experimental (n = 80) and control (n = 40) groups. Experimental animals received DMH 20 mg/kg s.c. for 20 weeks. Animals were fed either a regular diet (selenium content 0.2 p.p.m.) or a high selenium diet (5 p.p.m.) with or without 0.2% DFMO in the drinking water. At death, week 30, animal weights within experimental or control groups were not different between the four diet treatment groups. Tumor number and incidence in the proximal colon was not affected by DFMO treatment, selenium supplementation or the combined treatment. In contrast, in the distal colon, 19 tumors developed in the DFMO treated group, 22 tumors in the high selenium group and only 12 tumors in the combined high selenium/DFMO treatment group compared to 32 tumors in the regular diet group. Similarly, tumor incidence was decreased by DFMO and selenium supplementation and their effects were additive. In control animals, ODC activity was decreased by DFMO treatment and selenium supplementation in the distal colon and liver, but not the proximal colon. ODC activity of tumor tissue was greater than normal colon tissue from diet paired animals for proximal and distal colon, except for distal colonic tumors in the high selenium/DFMO treatment group. Polyamine content, however, did not correlate with ODC activity in normal or neoplastic tissue. In general, S-adenosylmethionine levels from normal colon and liver tissue were unaffected by diet treatment. Selenium supplementation in combination with DFMO treatment selectively inhibited distal colon tumor formation in rats fed a fiber-free diet.

Effect of selenium on growth, S-adenosylmethionine and polyamine biosynthesis in human colon cancer cells.

Selenium metabolism and polyamine biosynthesis are linked in their common requirement for S-adenosylmethionine. The effects of selenium supplementation (0.1 to 6.0 ppm) on growth, polyamine biosynthesis and S-adenosylmethionine were examined in two human colon cancer cell lines, WiDr and HT29. WiDr cells were very sensitive to selenium with a significant decrease in 3H-thymidine incorporation and cell number to doses above 0.25 ppm. HT29 cells were less sensitive. In HT29 cells, ornithine decarboxylase activity and its product putrescine decreased in parallel with the growth inhibitory effects of selenium. Similar changes were not noted in WiDr cells. Spermidine and spermine content were conserved in both cell lines exposed to cytotoxic doses of selenium. S-adenosylmethionine was increased in HT29 cells at cytotoxic doses of selenium (1.0 to 6.0 ppm).

Effects of fat and fiber on human colon cancer xenografted to athymic nude mice.

The effects of unsaturated fat and fiber (cellulose) on the growth of human colon cancer explanted to athymic nude mice was evaluated. Eighty-seven male nude mice bearing xenografts of human HT29 or WiDr colon cancer were divided into three groups of equal weight and tumor volume. Each group was fed one of three diets: normal fat/no fiber (N/N), high fat/no fiber (H/N) or high fat/high fiber (H/H). To equalize caloric intake, animals in the H/N group received 4 g of food per day and the other animals were fed 5 g of food per day. At sacrifice tumor volume and weight was recorded, and tumors were analyzed for protein and DNA content and ornithine decarboxylase activity. Tumor volume, weight, and protein were greater in the H/N group compared to the N/N group for both colon cancer cell lines. Tumor DNA content was greater in the HT29 H/N group compared to the N/N group (P less than 0.05) and tumor ornithine decarboxylase activity in the WiDr H/N group was greater than the N/N animals (P less than 0.002). The tumor growth-promoting effects of the high unsaturated fat diet were attenuated by the addition of fiber. Animal weight was higher in the H/N group compared to the N/N and H/H groups. This study suggested that a high-fat diet stimulated and fiber decreased the growth of human colon cancer explanted to athymic nude mice. The growth-promoting effects of a high-fat diet in colorectal cancer may be due in part to a circulating trophic factor since these tumors were remote from the large intestine.

Interspecies comparison of calmodulin binding proteins throughout the gastrointestinal tract: comparison with human colon adenomas and adenocarcinomas.

Calmodulin is an ubiquitous cytoplasmic protein which mediates many of the actions of calcium on intestinal tissue including regulation of growth and differentiation of normal and neoplastic cells. Using a biotinylated calmodulin overlay system, we compared the pattern of calmodulin binding proteins throughout the gastrointestinal tract of mice, rats, rabbits, and humans, and in human colonic adenomas and adenocarcinomas. A common calmodulin binding protein of 67 kDa was found in membrane and cytosolic fractions of oesophagus, stomach, proximal and distal small intestine, and colon from all four species. In human tissue this 67 kDa protein was present in greatest concentration in stomach tissue. Furthermore, a 67 kDa binding protein was the major calmodulin binding protein from human stomach and ileum as determined by ion exchange and calmodulin affinity chromatography. A similar pattern of binding proteins was noted between rabbit and human cytosolic fractions; proteins of 60/67 kDa and 105 kDa were present in stomach tissue. A 94 kDa protein was present in samples of rabbit and human ileum but not of mouse or rat. A similar pattern of calmodulin binding proteins was seen in normal and neoplastic large bowel tissue, apart from one of nine adenocarcinomas, where a distinct 54 kDa band was noted in both cytosolic and membrane fractions. The results of this study show interspecies and organ differences between calmodulin binding proteins, but suggest that a 67 kDa protein is the major binding protein present throughout normal gastro-intestinal tract and neoplastic human tissue.

Alterations in lectin binding in the proximal and distal colon of Sprague-Dawley rats with 1,2 dimethylhydrazine administration.

Sequential changes in rhodamine or fluorescein isothiocyanate-conjugated lectin binding of proximal and distal colonic crypts were studied during and after the administration of the 1,2 dimethylhydrazine (DMH). Five adult its unexposed to DMH or vehicle served as baseline controls. Tissue from normal appearing colon and tumor tissue was incubated with Ulex europaeus agglutinin 1 (UEA), Arachis hypogaea (PNA), Dolichos biflorus (DBA) and Griffonia simplicafolia 1 (GSA1). Distinct regional differences were noted in the baseline controls. UEA, PNA and DBA binding were absent in the distal colonic crypt cells. In the proximal colon minimal UEA and PNA binding was noted in the lower crypt whereas DBA binding was intense. GSA1 binding was diffuse in the upper and lower crypt of both regions. During carcinogenesis a progressive increase in PNA binding was noted in normal appearing colonic crypts from both regions. A progressive increase in PNA binding in proximal and distal colonic tumors was noted over time. Similar to normal tissue, DBA bound markedly to proximal colon tumors but was absent in most distal colonic tumors. UEA stained all proximal tumors intensely at all time points. In distal colonic tumors, UEA staining was diminished at 30 weeks compared to tumors analyzed at 16, 22 and 26 weeks. Mucin depletion was also a feature of tumor tissue compared to adjacent normal and hyperplastic glands. This study documents the region specific changes in lectin binding in normal and neoplastic colonic crypts induced by DMH.

Colonic polyamine content and ornithine decarboxylase activity as markers for adenomas.

Polyamine content (putrescine, spermidine, and spermine) or ornithine decarboxylase (ODC) activity was measured in normal-appearing colonic mucosa from patients undergoing colonoscopy. Comparisons were made between those with and those without adenomatous polyps. Colonic mucosal polyamine content was measured in 44 persons. Mean putrescine content was 1.25 +/- 0.26 (SE) nmol/mg protein in 22 patients with adenomatous polyps compared with 0.53 +/- 0.12 nmol/mg protein in patients without polyps (P less than 0.02). Tissue content of spermidine and spermine did not differ between these two groups. Ornithine decarboxylase activity was measured in tissue from 45 patients. Mean ODC activity was 2.84 +/- 0.73 pmol/hr/mg protein in 23 persons with adenomatous polyps compared with 1.15 +/- 0.18 pmol/hr/mg protein in persons without polyps (P less than 0.05). Mucosal putrescine and ODC activity are elevated in patients with adenomatous polyps compared with patients without polyps. These biochemical markers may prove helpful in improving surveillance methods for colorectal cancer and premalignant adenomatous polyps.

Results of a repeat television-advertised mass screening program for colorectal cancer using fecal occult blood tests.

The results of a 1987 television-advertised colorectal screening program using fecal occult blood tests (FOBT) are compared with the initial 1986 program (results in parentheses). In the 1987 program, 73,508 fecal occult blood test (FOBT) kits were distributed free of charge, of which 63% were returned for analysis (57,000, 53%). Twenty-five percent of persons from the initial screening participated again in the 1987 program: 1,303 or 2.8% of persons had a positive screen (1,165, 3.9%). The predictive value of a positive screen was 23% for an adenomatous polyp and 8% for colorectal cancer (22%, 8%). Seventy-nine percent of the cancers detected were Dukes A or B or carcinoma in situ (78%). In order to promote a more thorough diagnostic work-up in positive screenes, a suggested diagnostic algorithm for the work-up of a positive FOBT was sent to participating physicians. Despite this, 35% of positive screenees had a diagnostic work-up limited to a repeat FOBT, and/or sigmoidoscopy only (32%). In conclusion, television-advertised mass screening programs consistently enroll large numbers of participants. The rate of compliance (percent of kits returned) and the limited diagnostic evaluation of persons with a positive screen appear to be the major factors limiting the success of our screening program.

Lectin histochemistry of adenomatous polyps. Not a predictor of metachronous lesions.

The binding characteristics of fluorescein isothiocyanate conjugated lectins to normal colonic mucosa, and 43 adenomatous polyps were studied by fluorescence microscopy. The lectin, Dolichos biflorus agglutinin (DBA) stained intensely to upper crypt cells of the sigmoid colon and rectum but to a lesser degree to proximal colonic crypts or lower crypt cells distally. Peanut agglutinin (PNA) and Ulex europaeus agglutinin (UEA) did not bind to the theca of proximal or distal colonic crypts. The lectin Griffonia simplicafolia agglutinin (GSA1) bound intensely to upper and lower crypt cells of both regions. PNA binding was noted in 56% of adenomatous polyps, occurred more often in polyps of the distal colorectum, and increased with polyp size and villous histology. UEA bound to 26% of adenomatous polyps, 42% of proximal polyps, and 17% of distal polyps. DBA staining was noted in 72% of polyps without regional preference. GSA1 stained all polyp specimens. To determine if the lectin binding characteristics of an index (initial) polyp might serve as a predictor of metachronous lesions, 20 patients (29 polyps) without a history of polyps or cancer and who had at least one surveillance colonoscopy 1 to 3 years after the initial polypectomy were studied. The presence or absence of PNA, UEA, or DBA binding in an index polyp did not predict the occurrence of metachronous lesions. Five of the six patients with more than one index polyp had metachronous polyps at follow-up surveillance colonoscopy.

Structural and enzymatic changes during colonic maturation in the fetal and suckling rat.

To assess correlations between cellular differentiation and enzymatic maturation in the developing rat colon, tissue from fetal, suckling, weanling, and adult rats was analyzed by electron microscopy and assayed for lactase, alkaline phosphatase, and sodium-potassium-stimulated adenosine triphosphatase activities. The proximal and distal colon were analyzed independently at all ages. All three enzymes were detected in the fetal colon when the cells were highly undifferentiated. Postnatally, significant regional differences in cellular ultrastructure appeared, only some of which were directly paralleled by enzymatic changes. Each enzyme had a distinct region-specific developmental pattern. Lactase and sodium-potassium-stimulated adenosine triphosphatase were significantly enhanced at birth, decreasing to adult levels by 15 days postnatal. Regional differences were present, but the patterns were similar. These patterns did not parallel the increase in microvillar height and number and basolateral interdigitations of the surface columnar cells, the structural correlates of lactase, and sodium-potassium-stimulated adenosine triphosphatase, respectively. In contrast, developmental changes in alkaline phosphatase activity paralleled structural maturation, at least in part. The activity levels in the distal colon did not change significantly with age and few major structural changes were noted. In the proximal colon, activity increased markedly after birth, and after 10 days decreased rapidly to adult levels, a pattern that coincided with the transient appearance of villi and specialized cells with apical tubules and vesicles known to have alkaline phosphatase activity. The results show age- and region-related changes in cellular ultrastructure and enzymatic activities, only some of which appear to be directly correlated.

Results of a television-advertised public screening program for colorectal cancer.

We report the results of a free, television-advertised mass screening program for colorectal cancer using stool guaiac kits. A total of 57,000 test kits were picked up and 29,619 (53%) were returned; 3.9% (1165) of the tests were positive. Ninety-three percent of persons with a positive screen sought medical evaluation after screening. Detailed follow-up was available on 744 persons. Fifty-eight persons had large-bowel carcinomas diagnosed, 80% of which were localized. One hundred sixty persons had adenomatous polyps removed. Forty percent of cancers and 58% of polyps were detected in persons with only one or two positive test slides out of a total of six. In 33% of persons with a positive screen, the diagnostic workup consisted of a repeated stool guaiac test and/or sigmoidoscopy only. A major drawback to improving the results of mass screening programs for colorectal cancer is the limited gastrointestinal workup conducted by physicians in many persons with a positive fecal occult blood test.

The effects of chronic ethanol administration on polyamine content during dimethylhydrazine-induced colorectal carcinogenesis in the rat.

Chronic ethanol (EtOH) consumption has been implicated as a co-carcinogen, selectively promoting rectal tumor formation. We studied the effects of EtOH consumption on tumor formation and polyamine content (putrescine, spermidine and spermine) in proximal and distal colon and rectum of Sprague-Dawley rats treated with the procarcinogen 1,2-dimethylhydrazine (DMH). Sixty-four adult male rats were pair fed nutritionally complete liquid diets with 36% of calories supplied as EtOH or isocaloric carbohydrates. Both groups received 4 weeks of the liquid diet followed by 4 weeks of standard laboratory chow during which 50% of the rats in each group received DMH (30 mg/kg) or vehicle s.c. weekly. This cycle was repeated four times (32 weeks). Animals were sacrificed at the end of each 8 week cycle and normal appearing and available tumor bearing tissue from proximal and distal colon and rectum was obtained for polyamine content and histology. Five animals, unexposed to DMH or EtOH served as baseline controls. There were no consistent regional differences in putrescine, spermidine or spermine of baseline controls. A progressive decrease in tissue putrescine was seen in all three regions of the control group and was significant at 24 and 32 weeks versus baseline controls. In all three regions, chronic EtOH consumption prevented the decrease in tissue putrescine. Spermidine content was also significantly increased in the distal colon of EtOH-treated animals compared to baseline values. Consistent changes in spermine content were seen in any treatment group or region. A significant increase in putrescine content of normal appearing and tumor-bearing tissue of DMH treated animals at 32 weeks was noted. Chronic EtOH administration did not augment rectal or colonic polyamine content in DMH-treated animals. Likewise, chronic EtOH consumption did not alter the number, size or distribution of large bowel tumors of DMH treated animals.

Cellular proliferation in proximal and distal rat colon during 1,2-dimethylhydrazine-induced carcinogenesis.

Sequential changes in proliferative parameters in proximal and distal colonic crypts were studied during 1,2-dimethylhydrazine-induced carcinogenesis using [3H]thymidine autoradiography as a probe. 1,2-dimethylhydrazine (20 mg/kg) and vehicle (ethylenediaminetetraacetic acid) control rats received weekly s.c. injections for 20 wk. All animals received a pulse of [3H]thymidine before death at weeks 2, 6, 10, 16, 22, 26, or 30. In addition, 8 animals unexposed to 1,2-dimethylhydrazine or vehicle served as baseline controls. Dramatic regional differences were noted in the baseline controls. Crypt length, labeling index, and proliferative zone size were all significantly greater distally than proximally (p less than 0.05), whereas the labeling index of the proliferative zone tended to be enhanced proximally. During 1,2-dimethylhydrazine treatment the crypt length, labeling index, and proliferative zone size increased in both regions. As these parameters changed in parallel, the differences between proximal and distal colon did not change significantly during carcinogenesis. Actual tumor formation did differ, however, with tumors appearing earlier and in greater abundance in the distal colon. These findings show similar proliferative changes in both the proximal and distal colon during 1,2-dimethylhydrazine treatment and indicate that the enhanced baseline proliferative state of the distal colon compared with the proximal colon must be considered in the process of tumor formation.