Changes in plasma, erythrocytes and tissue polyamine levels in patients with colorectal cancer

Polyamines (putrescine, spermidine, spermine) are closely linked to cellular synthesis of DNA, RNA and protein, and are thought to be an indicator of cell proliferation. Plasma, erythrocytes and tissue polyamine levels in 58 patients with colorectal cancer were measured to survey the relationship between polyamines and stage classification. The polyamine levels of plasma, erythrocytes and tissue in patients were increased significantly compared with those of controls. Plasma spermine and erythrocytes spermidine and spermine levels were increased with the advance of stage. In plasma and erythrocytes, spermidine/spermine ratios were decreased in accordance with the stages. On the contrary, the polyamine levels and the ratio in cancer tissue were not varied in all stages. These results present the following conclusions. Although cancer tissue has higher proliferative activity than normal mucosa, these activities of main tumors in each stage are not fluctuated. The polyamine levels in plasma and erythrocytes are possibly influenced by tumor burden and therefore those could be an useful marker for indicating the stage of colorectal cancer.     

High ornithine decarboxylase activity and polyamine levels in human colorectal neoplasia.

Polyamines are required for cell proliferation, and ornithine decarboxylase (ODC) is the first and probably rate-limiting enzyme in their synthesis. Tissue containing colonic or rectal adenocarcinomas (N = 34) or polyps (N = 6) and noninvolved paired colonic mucosa were obtained from fresh surgical specimens. ODC activity was elevated (mean: 320%) in both the cancer and polyps. In noninvolved colonic mucosa of tumor-bearing specimens, ODC activity was 165% that of colonic mucosa of non-neoplastic disease. Concentrations of polyamines in neoplasms were 121-214% increased, as compared with normal mucosa; those of spermidine and spermine varied inversely with the histological grade of the tumor. High levels of ODC activity and of polyamines were features of neoplasia, but not of malignancy alone. These characteristics of colonic neoplasia suggest its susceptibility to control by inhibition of ODC.     

Detection of polyamines by an enzymatic assay. (6). Fundamental and clinical studies of a simple enzymatic method for determining total polyamines in blood

A simple enzymatic method for determining total polyamines in human blood was established. Polyamines in trichloroacetic acid extract from 1 ml of blood were isolated on an anion-exchange column and measured spectrophotometrically by the end point assay using polyamine oxidase and putrescine oxidase. The recovery was as high as 98.4% and within-run precision (coefficient of variance: 1.82%), and the values obtained by this method were in fair agreement with those obtained by high performance liquid chromatography and the enzymatic differential assay methods previously reported. The polyamine levels were not changed when the blood was stored at -20 degrees C. Blood polyamine levels were measured in 108 patients with genitourinary cancers, 29 patients with benign prostatic hypertrophy, 18 patients with benign urological diseases and 25 normal subjects. Although polyamines were not significantly elevated in the low stage of cancer, elevation was observed at a high stage of malignancy. These results indicate that the determination of total polyamines may be useful to determine biochemically the malignancy of cancers.     

Polyamines in colorectal cancer

Polyamine levels (putrescine, spermidine and spermine) in colorectal cancers (n = 25) were measured in order to assess their importance as markers of cellular proliferation. Colonic mucosa from healthy resection margins of patients with diverticular disease (n = 5) was used as control material. Polyamine levels (expressed as nanomoles per 100 mg tumour) in cancers ranged from 0.8 to 7.9 for putrescine (mean: 2.3 +/- 0.7), from 6.5 to 22.8 for spermidine (mean: 13.9 +/- 0.9) and from 13.0 to 37.5 for spermine (mean: 22.1 +/- 1.3). Mean spermidine and spermine content of cancers was more than three times mean spermidine (3.92 +/- 0.8), and more than four times mean spermine (5.0 +/- 1.2), content of normal colonic mucosa (P less than 0.01). Polyamine content of colorectal cancers was independent of tumour site, Dukes' stage, histological grade and the presence of palpable liver metastases at laparotomy. Because colorectal cancers contain such high levels of spermidine and spermine, polyamines may play an essential role in the regulation of their growth.     

Polyamines in breast cancer

Polyamine levels (putrescine, spermidine and spermine) in breast cancers (n = 54) were measured as a potential guide to prognosis. Values (expressed as nmol per 100 mg tumour) ranged from: 0.9 to 4.5 for putrescine, 4.2 to 29.8 for spermidine and 5.6 to 39.7 for spermine concentration. Increased intracellular polyamine levels were positively correlated with factors known adversely to affect survival after mastectomy, namely histological grade III and oestrogen-receptor negative status. Advanced T4 tumours and medullary-type carcinomas also contained high polyamine levels. Tumour size and node status did not affect polyamine levels in primary tumours. Tumours that recurred within 2 years of mastectomy had significantly higher levels of spermidine and spermine than those that did not. Breast cancer polyamine levels are a biological marker of tumour aggressiveness and can be used as a prognostic indicator of early tumour recurrence that is independent of node status.     

Differential release of polyamines by cultured rat Sertoli cells

Cellular and media concentrations of polyamines in Sertoli cell cultures were determined by fluorescent spectroscopy of dansylated compounds after separation by high-performance liquid chromatography. In spite of low cellular levels of putrescine, the Sertoli cells released relatively large amounts of putrescine and spermidine even after several media changes. The inclusion in the culture media of cortisol, insulin, and thyroxine significantly elevated cellular polyamine levels, altered the spermidine to spermine ratio, and enhanced putrescine release by 3- to 4-fold. No spermine, however, was detected in the media under any of the conditions studied. The polyamine concentrations in cultured Sertoli cells from 13-day-old rats and the pattern of polyamine release by these cells differed significantly from those in the Sertoli cells from 46-day-old rats. These data demonstrate the differential release of polyamines by cultured rat Sertoli cells. The profiles of polyamine secretion appear to be age-dependent, and the significance of this phenomenon is discussed.     

Detection of polyamines by a new enzymatic differential assay. (8) Studies on tissue polyamine concentrations in patients with genitourinary malignant diseases

Polyamine concentrations of human cancerous and non-cancerous tissues from the kidney, ureter, bladder were measured by a new enzymatic method for isolation and determination of polyamines. In cancerous and non-cancerous tissue of the organs studied, the spermine level was highest followed by the spermidine and diamine levels. The concentrations of diamine, spermidine and spermine in cancerous tissues were significantly higher than those in non-cancerous tissues, but there was no significant difference in the spermidine/spermine ratio between the cancerous and non-cancerous tissues. These data suggest that polyamines are produced above the normal levels in pathological conditions such as renal cell carcinoma, ureteral cancer and bladder cancer.     

Polyamine metabolism in carcinoma of the oral cavity compared with adjacent and normal oral mucosa

In this study, polyamine biosynthesis required for cellular proliferation showed elevated levels in neoplastic cells. Putrescine, spermidine, and spermine, as well as the rate-limiting enzyme ornithine decarboxylase, were measured to evaluate differences in tissue concentration in squamous cell carcinoma of the oral cavity, and in the normal adjacent, buccal, and retromolar trigone tissues. Mean polyamine levels (nanomoles per gram of tissue +/- standard error of the mean) were significantly elevated in tumor tissue at 136 +/- 42 nmol/g for putrescine compared with 41 +/- 9 nmol/g in adjacent, 25 +/- 5 nmol/g in buccal, and 41 +/- 14 nmol/g in retromolar trigone tissues. Tumor spermidine was 415 +/- 41 nmol/g compared with 192 +/- 34 nmol/g in adjacent, 184 +/- 34 nmol/g in buccal, and 214 +/- 63 nmol/g in retromolar trigone tissues. Tumor spermine was 461 +/- 41 nmol/g compared with 236 +/- 30 nmol/g in adjacent, 233 +/- 35 nmol/g in buccal, and 269 +/- 59 nmol/g in retromolar trigone samples. Ornithine decarboxylase activity was highly variable in tumor tissues. High levels of polyamines appear to be specific for this malignancy, whereas ornithine decarboxylase activity is not. Measurement of polyamine content may be useful in evaluating epithelial changes in the oral cavity.     

The clinical value of urinary polyamine analysis in urological disease

To study the clinical usefulness of the determination of urinary polyamine levels, voluntary urine of several urological diseases including 56 bladder tumor patients was analyzed by high performance liquid chromatography. The obtained values were adjusted by the concentration of urinary creatinine and expressed as the unit of mumol/g creatinine (mumol/g Cr) From the measurement of 8 normal adults, the normal upper limit of each polyamine was decided by mean + 2SD, and the limit for total polyamine was 59.1 mumol/g Cr, putrescine 38.1 mumol/g Cr, spermidine 16.6 mumol/g Cr and spermine 9.2 mumol/g Cr, respectively. In the patients with non-neoplastic benign urological disease, the polyamine levels were statistically not different from those of the normal adults. In the case of bladder tumor, the urinary levels of total polyamine, putrescine and spermine were significantly elevated compared with the control group. The true positive rate of this determination in bladder tumor patients was 26/56 (46%) by total polyamine level, 21/56 (38%), by putrescine level, 11/56 (56%) by spermidine level and 16/56 (29%) by spermine level. Grade or stage of the bladder tumor did not have any significant correlation with the urinary polyamine level. This determination would not be included in routine clinical examinations due to the difficulty of measurement, difference of urine sampling and lack of high sensitivity and specificity.     

Effect of polyamines on mesangial cell ecto-5'-nucleotidase and ecto-ATPase activity

Polyamines were found to modulate the activity of several membrane-bound enzymes, participating in cell growth and differentiation. We have studied the effect of polyamines (spermidine, spermine and putrescine) on rat mesangial cell ectoenzymes: 5'-nucleotidase, Mg(2+)-ATPase and Ca(2+)-ATPase. Ecto-5'-nucleotidase activity was significantly increased after 48 h treatment with spermine and spermidine. Mg(2+)-ATPase was increased only after treatment with spermidine; however, Ca(2+)-ATPase was significantly increased after both spermine and spermidine treatment of mesangial cells. Culture of mesangial cells with putrescine did not change the activity of these ectoenzymes. Increased expression of mesangial cell ecto-ATPase and ecto-5'-nucleotidase after spermine and spermidine treatment could result in an increased production of adenosine, a powerful autacoid interesting with respect to a role of mesangial cells in inflammatory processes.     

Calmodulin antagonist trifluoperazine inhibits polyamine biosynthesis and liver regeneration

Polyamines are essential for cell growth and differentiation. Trifluoperazine (TFP) is a potent, competitive inhibitor of the calcium-calmodulin complex. TFP, when given to rats after partial hepatectomy, causes a significant decrease in DNA synthesis. The purpose of this study was to examine the effects of TFP on polyamine biosynthesis and on liver regeneration after partial hepatectomy. TFP (60 mg/kg, bodyweight) or saline control was administered to 80 male Sprague-Dawley rats 2 h before, 2 h after, or at the time of hepatectomy. Polyamines (putrescine, spermidine and spermine) were measured at the time of hepatectomy, and at 6, 24, 48, and 72 h after hepatectomy. TFP, when it was administered either 2 h before or at time of hepatectomy, blocked increases in putrescine that are seen normally at 6 h after hepatectomy. When TFP was given at the time of partial hepatectomy, putrescine was increased at 24 h, and then returned to normal levels at 72 h. Spermidine was inhibited at 24 h, but not at 48 and 72 h. Spermine was not significantly altered at any time. The administration of TFP 2 h after hepatectomy did not significantly alter concentrations of polyamines. The weight of regenerating liver was decreased by TFP at 48 h (23 per cent) and 72 h (22 per cent) after hepatectomy. These findings provide evidence that the calcium-calmodulin complex is required for the synthesis of liver polyamines before liver regeneration can proceed.     

Altered polyamine concentrations in cytomegalovirus-infected human cells

Polyamines have a regulatory effect on DNA and RNA synthesis and their levels are elevated in rapidly growing cells, including lymphoblasts. However, as shown in the current experiments, exposure to cytomegalovirus (CMV) reduces the polyamine levels in these cells, suggesting that the virus interferes with their metabolism. Studies have shown that the activity of ornithine decarboxylase is increased in CMV-infected fibroblasts and that there is an increased conversion of putrescine to spermidine and spermine. Thus it may be expected that the concentration of these molecules would increase in the infected cell. However, the results presented here demonstrate that only the concentrations of putrescine and spermidine are increased, the spermine concentration decreasing with infection.     

Change in serum polyamine metabolome pattern after bariatric surgery in obese patients with metabolic syndrome

BackgroundRecent works have reported that bariatric surgery has remarkable effects on the metabolome, which might be potentially associated to the metabolic improvement of this procedure in patients with obesity. Serum polyamines, metabolites derived from amino acid metabolism, have been recently related to the metabolic status in obese individuals. However, the impact of bariatric surgery on the circulating levels of polyamines remains elusive.ObjectiveTo evaluate the effect of bariatric surgery on serum polyamine levels and to evaluate the association of changes in these molecules with metabolic improvement in patients with morbid obesity.SettingVirgen de la Victoria University Hospital, Malaga, Spain.MethodsThis study included 32 morbidly obese patients (weight index ≥40 kg/m²) with metabolic syndrome, who underwent sleeve gastrectomy. Serum levels of polyamines (putrescine, spermidine, and spermine), acetylpolyamines, and polyamine-related amino acids (arginine and ornithine) were assessed at baseline and 6 months after bariatric surgery, and were analyzed in an ultraperformance liquid chromatography–mass spectrometry platform.ResultsOur metabolomic analysis revealed a significant rise in several metabolites related to the polyamine metabolism, such as putrescine and acetyl derivatives of spermidine and spermine in serum samples from morbidly obese patients after bariatric surgery. Changes in serum levels of both putrescine and acetylputrescine were associated to the resolution of metabolic syndrome after surgery.ConclusionOur study indicates that bariatric surgery affects the serum polyamine pattern and the resolution of metabolic syndrome after bariatric surgery is associated to specific changes in the serum polyamine metabolome.     

Determination of blood polyamines in patients with brain tumor -with special reference to relationship between varieties of tumors and concentrations of blood spermidine and spermine (author's transl)]

Examination of blood polyamines in 38 patients with brain tumor and 17 normal volunteers was carried out by columnar chromatography--cellulose acetate membrane electrophoresis. The upper limits of the normal values; M.+2S.D. of the blood polyamine concentrations in 17 normal volunteers, were less than 2.1 mg/ml for spermidine, less than 1.6 mg/ml for spermine, and less than 2.2 mg/ml for spermidine plus spermine. The values of blood polyamines in 21 cases with glioma were significantly higher than those in normal subjects (p less than 0.01). And in 14 out of them, the concentrations of the blood polyamines were higher than the maximum normal value. In one case with reticulum cell sarcoma, the concentrations of the blood polyamines were remarkably increased. In 2 out of 4 cases with metastatic brain tumor the concentration of the blood polyamines were higher than the upper limit of normal amount, and values of the blood polyamines in 4 cases with metastatic brain tumor were significantly higher than those in normal volunteers (p less than 0.05). In none of 2 cases with pituitary adenoma, 3 cases with meningioma, 4 cases with neurinoma, one case with hemangioblastoma, and one case with pinealoma, the values of the blood polyamines were significantly higher than those in normal volunteers. The CSF samples obtained from 9 patients with brain tumor, consisted of 6 gliomas (glioblastoma multiforme 2, anaplastic glioma 4), 1 teratoblastoma, 1 von Recklinghausen's disease (neurinoma and meningioma), and 1 craniopharyngioma, were analyzed for detection of polyamines, but no detectable amount was present in those cases.     

The clinical significance of tissue, blood and urine polyamine in renal cell carcinoma]

With the use of a newly developed and convenient enzymatic method, tissue, urine and blood polyamine (diamine, spermidine and spermine) levels were evaluated as a tumor marker of renal cell carcinoma (RCC) in 50 cases with the disease. Furthermore, blood and urine polyamines were periodically determined and evaluated as a follow-up marker. The pretreatment three polyamine levels in tissue, blood and urine of the patients were all significantly higher than those of the controls. However, because of their low sensitivities, they were not always decisive for biochemical diagnosis of RCC. Tissue spermidine levels were increased with the advance of the stages. Tissue diamine level also showed a good correlation with the pathological grade. Tissue diamine was found to predict distant metastasis. Blood spermidine and urine diamine were useful as follow-up markers. In conclusion, combined determination of tissue, blood and urine polyamine levels was thought to be useful as tumor markers of RCC.     

Polyamines in the Gastro‐intestinal Tract of Goat Kids and in the Regenerating Ruminal Epithelium of Sheep

The aim of the present study was to investigate whether temporal changes in polyamine concentration and synthesis could be found in the luminal content and wall tissue of the rumen and abomasum, two organs which have entirely different growth patterns during the first month of life. In the abomasal mucosa there was a marked gradual decrease in the ornithine decarboxylase (ODC) activity during the first month of life, while the ODC activity in the ruminal mucosa was low during the whole experimental period. However, injury of the rumen wall was followed by increased ODC activity. The ODC activity in duodenal mucosa was about 10 times higher than in the ileal mucosa and the ruminal epithelium. In ruminal liquid a clear peak in ODC activity was observed during the period 51–70 days after birth. The polyamine concentration did not parallel the ODC activity, in either the ruminal epithelium or the ruminal liquid. Of the polyamines, the spermine concentration was always highest, and with the exception of duodenal mucosa, the putrescine concentration was lowest. In liver a clear decrease in spermidine concentration from day 1 to about day 60 after birth was observed. Otherwise no marked temporal changes in tissue polyamine concentrations were observed. Two and a half hours after oral administration of ¹⁴C‐labelled spermine, nearly all of the radioactivity was found in the lumen of the gastrointestinal tract. On the other hand, 1 h after intravenous injection of polyamines the walls of the gastrointestinal tract were strongly labelled. In conclusion, the polyamines needed for ruminal epithelial development seem to come from sources other than the ruminal epithelium itself or the ruminal lumen.     

Ornithine decarboxylase activity as a biochemical marker in individuals predisposed to lung cancer

Ornithine decarboxylase (ODC) is the first enzyme in the pathway of mammalian polyamine biosynthesis. "Tumor promoters" appear to induce ODC. In addition, increased ODC activity has been observed in normal appearing colonic mucosa of patients with familial polyposis, an autosomal dominant disorder associated with a high incidence of colon cancer. Ornithine decarboxylase activity was measured in bronchoscopic mucosal biopsy specimens from the noninvolved lung in patients with unilateral lung masses. No correlation could be determined in ODC levels from "normal" mucosa between patients with lung cancer compared to those with benign disease, despite elevated ODC activity in the tumors themselves.     

Urine levels of polyamine in patients with or without cancer

The urine levels of polyamine (total amount of putrescine, spermidine and cadaverine) were measured in patients with or without cancer by simple enzymatic assay method. In 148 control healthy adults, the urine levels of polyamine were 23.1 +/- 7.1 mumole/g. creatinine, whereas among 52 patients with benign diseases, the level in only 5 patients was slightly higher than normal level. The polyamine levels in 170 patients with cancer was 46.1 +/- 50.6 mumole/g. creatinine, which was about 2-times higher than normal level. In the patients with cancer of the stomach or the colon and rectum, the increase in polyamine level appeared to be correlated with the clinical stage of tumor. Following successful surgical resection of cancer, the polyamine level increased in one week transitorily after operation but gradually decreased to normal level within 5 weeks. Whereas following unsuccessful surgical resection of cancer, the polyamine value maintained high levels. The evidence suggests that the measurement of the urine level of polyamine is useful for the diagnosis of cancer or the clinical stage of tumor, and it will be helpful in the evaluation of therapeutic effects and prognosis.     

Detection of urinary polyamine by a new enzymatic differential assay. (I). Fundamental study on a new enzymatic differential assay

A new enzymatic method for determining urinary polyamine concentration by fractionation of the urinary acetyl conjugate into free polyamines with acylpolyamine amido-hydrolase and quantification using two kinds of amine-oxidase of different substrate specificity was examined. High recovery rates of urinary polyamine by enzymatic hydrolyzation were obtained, namely, 95 +/- 4% for diamine, 95 +/- 1% for spermidine and 99 +/- 2% for spermine. Furthermore, excellent linearity was demonstrated with up to 150 mumole/l diamine, up to 75 mumole/l spermidine and up to 50 mumole/l spermine. Although urinary polyamine concentration varied diurnally even after correction of urinary creatinine, day-to-day variation disappeared. In 24-hour pooled urine and voluntary urine, diamine, spermidine and spermine correlated relatively well. Urinary leukocytes and erythrocytes exerted no influence on urinary polyamine.     

大肠癌中胃泌素和胃泌素mRNA的检测

目的研究大肠癌组织胃泌素和胃泌素ｍＲＮＡ的表达水平,探讨胃泌素在大肠癌生长中的作用。方法应用放射免疫法检测４０例大肠癌组织和相应的癌旁正常粘膜组织胃泌素,同时应用半定量ＲＴ－ＰＣＲ方法对胃泌素ｍＲＮＡ表达水平进行分析。结果大肠癌组织胃泌素、胃泌素ｍＲＮＡ表达水平明显高于正常对照组（Ｐ＜０．０５）,同时发现胃泌素、胃泌素ｍＲＮＡ表达水平和大肠癌的Ｄｕｋｅｓ分期及转移成显著正相关。结论证实胃泌素具有自分泌促肿瘤生长作用,且胃泌素表达的增加很可能发生在肿瘤中晚期,可作为大肠癌预后的重要指标。