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Doi:10.1016/j.biocel.2004.08.002The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 Insulin in aging and cancer: antidiabetic drug diabenol as geroprotector and anticarcinogen Irina G. Popovich, Mark A. Zabezhinski, Peter A. Egormin, Margarita L. Tyndyk, Ivan V. Anikin, Alexander A. Spasov, Anna V. Semenchenko, Anatoly I. Yashin, Vladimir N. Anisimov a Department of Carcinogenesis and Oncogerontology, N.N. Petrov Research Institute of Oncology, St. Petersburg 197758, Russia b Department of Pharmacology, Volgograd State Medical University, Volgograd, Russia c Max-Planck Institute for Demographic Research, Rostock, Germany d Center for Demographic Sciences, Duke University, Durham, NC, USA Received 18 May 2004; received in revised form 20 July 2004; accepted 4 August 2004 The effects of new antidiabetic drug Diabenol® (9-␤-diethylaminoethyl-2,3-dihydroimidazo-(1,2-␣)benzimidazol dihy- drochloride) on life span and spontaneous tumor incidence in NMRI and transgenic HER-2/neu mice as well as on coloncarcinogenesis induced by 1,2-dimethylhydrazine in rats are studied. It is shown that treatment with the drug failed influencebody weight gain dynamics, food and water consumption and the body temperature, slowed down age-related disturbances inestrous function and increased life span of all and 10% most long-living NMRI mice. The treatment with diabenol inhibited spon-taneous tumor incidence and increased the mammary tumor latency in these mice. Diabenol treatment slowed down age-relatedchanges in estrous function in HER-2/neu mice, failed influence survival of these mice and slightly inhibited the incidence anddecreased the size of mammary adenocarcinoma metastases into the lung. In rats exposed to 1,2-dimethylhydrazine, treatmentwith diabenol significantly inhibited multiplicity of all colon tumors, decreased by 2.2 times the incidence of carcinomas inascending colon and by 3.1 times their multiplicity. Treatment with diabenol was followed by higher incidence of exophytic andwell-differentiated colon tumors as compared with the control rats exposed to the carcinogen alone (76.3% and 50%, and 47.4%and 14.7%, respectively). Thus, the drug increases survival and inhibits spontaneous carcinogenesis in mice and inhibits coloncarcinogenesis in rats.
2004 Elsevier Ltd. All rights reserved.
Keywords: Diabenol; Antidiabetic drugs; Aging; 1,2-Dimethylhydrazine; Mammary carcinoma; Colon cancer; Carcinogenesis ∗ Corresponding author. Tel.: +7 812 596 8607; The potential link between aging and insulin/IGF-1 fax: +7 812 596 8947.
E-mail address: [email protected] (V.N. Anisimov).
signaling has attracted substantial attention during last 1357-2725/$ – see front matter 2004 Elsevier Ltd. All rights reserved.
I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 years, on the basis of evidence including age-related The concept of CR mimetics is now being in- increase in incidence of insulin resistance and type 2 tensively explored (; diabetes in accelerated aging syndromes as well as life span extension by caloric restriction (CR) in rodents.
volve interventions that produce physiological and Concomitant reduction in plasma insulin and plasma anti-aging effects similar to CR. It was suggested to glucose levels, which implies increases sensitivity to use biguanide antidiabetics as a potential anti-aging insulin, emerges as a hallmark of increased longevity Hyperglycemia is an important aging factor involved The antidiabetic drugs, phenformin, in generation of advanced glycosylation endproducts buformin, and metformin, were observed to reduce hy- perglycemia and produce the following effects: im- There is evidence that hyperinsuline- proved glucose utilization; reduced free fatty acid uti- mia favors accumulation of oxidized protein by reduc- lization, gluconeogenesis, serum lipids, insulin and ing its degradation as well as facilitates protein oxida- IGF-1, and reduced body weight both in humans and tion by increasing steady-state level of oxidative stress experimental animals There are no available data on the effect of other than biguanides abetics with elevated glucose levels suffer many mani- antidiabetic drugs on life span of animals.
festations of accelerated aging, such as impaired wound healing, obesity, cataracts, vascular and microvascular damage (It is important to stress that synthesised in Rostov State University, and its hy- hyperinsulinemia is an important factor not only in ag- poglycemic activity was evaluated as 1.5 times more ing but also in the development of cancer ( effective than of maninil (glibenclamide) and equal to the effect of glyclazide (pioglitazone) in rats, rabbits It was shown that hypoglycemic In organisms ranging from yeast to rodents, both effect of diabenol included both pancreatotropic and calorie restriction and mutations in insulin/IGF-1 extrapancreatic pathways. Diabenol restores physio- signaling pathway extend life span logical profile of insulin secretion and decreases tissue resistance to insulin, prolongs hypoglycemic effect of insulin. It increases glucose utilization in glucose loading test in the old obese rats. It was suggested that Both approaches have diabenol influence insulin receptors in peripheral tis- some side effects. For example, calorie restriction in- sues. Diabenol increases uptake of glucose by isolated creases the level of serum glucocorticoids and de- rat diaphragm in vitro both without supplementation of creases resistance to infection ( insulin into the medium or with supplemented insulin.
Diabenol also decreases platelet and erythrocyte whereas genetic modifications on insulin/IGF-1 signal- aggregation and blood viscosity, inhibits mutagenic ing pathway cause obesity, dwarfism and cardiopul- effect of 2-acetylaminofluorene and has antioxidant monary lesions (). Reviewing the available data on the benefits and adverse effects of calorie restriction and genetic modifications, three categories of drugs Thus, these results suggest that like which may have potential to prevent or postpone age- biguanides diabenol has a potential to increase the life related diseases and extend life span: drugs that (1) span and inhibit carcinogenesis.
stimulate dwarf mutations, and therefore, decrease pi- In this paper, we present the results of experiments tuitary production of GH; (2) prevent IGF-1 release with diabenol on some ageing related biological pa- from the liver; or (3) decrease IGF-1 signaling by the rameters, survival and spontaneous tumorigenesis in fe- action on either extracellular or intracellular targets.
male NMRI and transgenic HER-2/neu mice as well as I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 on 1,2-dimethylhydrazine-induced colon carcinogene- Once every 3 months, simultaneously with weigh- sis in rats.
ing, the amount of drinking water and consumed foodwas measured and the rate of consumed food (grams)per mouse was calculated.
2. Material and methods
Once every 3 months, vaginal smears, taken daily for 2 weeks from the animals, were cytologically exam- 2.1. Animals ined to estimate the phases of their estrous functions. Inthe same period, rectal body temperatures of the mice One hundred female NMRI 2-month-old mice and were measured with an electronic thermometer, TPEM 39 female 2-month-old LIO rats (KMIZ, Russia). The animals were observed until their were obtained from the Animal Department of N.N.
natural deaths. The date of each death was registered, Petrov Research Institute of Oncology. Homozygous and the mean life span, the age at which 90% of the an- HER-2/neu transgenic mice obtained from Charles imals died, and the maximum life span were estimated.
River (Hollister, CA) by the Italian National ResearchCenter for Aging were housed and breed in the Labora- 2.3.2. HER-2/neu mice tory of Carcinogenesis and Aging. The mice were kept At the age of 2 months, HER-2/neu 57 mice were 5–7 in polypropylene cages (30 cm × 21 cm × 10 cm) randomly divided into two groups. All mice were in- and rats were kept 5 in cages (46 cm × 32 cm × 16 cm) dividually marked. One group of 28 mice were given under standard light/dark regimen (12 h light:12 h dark- diabenol in drinking water (0.1 mg/ml) 5 days a week ness) at temperature 22 ± 2 ◦C and received standard monthly until their natural deaths. Twenty-nine con- laboratory chow (and tap water trol mice were given tap water without diabenol. Fresh solution was prepared ex tempore three times a week.
Animals were checked daily by animal care person- The observation schedule and tests were the similar to nel and weekly by a veterinarian. The study was carried these in experiments with NMRI mice.
out in accordance with the regulations for ensuring thehumane treatment of animals under the approval of the 2.3.3. LIO rats Committee on Animal Research of the N.N. Petrov Re- Thirty-nine 3-month-old outbred LIO rats were ran- search Institute of Oncology.
domly subdivided into two groups. All rats were ex-posed weekly to five subcutaneous injections of DMH 2.2. Chemicals at a single dose of 21 mg/kg of body weight (calcu-lated as a base). In this regimen, the carcinogen in- Diabenol, 100% pure, was provided by Dr. V.A.
duced colon tumors in the majority of rats ( Anisimova, Ph.D., Physical Chemistry Research Insti- tute, Rostov-on-Don, Russia ( DMH was ex tempore dissolved in normal saline and 1,2-Dimethylhydrazine dihydrochloride (DMH) neutralized with sodium bicarbonate (pH 7.0). Addi- was provided by Sigma (USA), and was kept at tionally, 5 days a week, 20 rats of one group were given diabenol with tap water (0.1 mg/ml) starting from theday of the first injection of the carcinogen during 26 2.3. Experiment weeks whereas 19 rats of another group were not giventhe drug. The experiment was finalized 6 months after 2.3.1. NMRI mice the first injection of the carcinogen, and all rats were At the age of 2 months, NMRI mice were randomly sacrificied by ether vapor.
divided into 2 groups, 50 animals in each, and they wereindividually marked. One group of mice were given 2.4. Pathomorphological examination diabenol in drinking water (0.1 mg/ml) 5 days a weekmonthly until their natural deaths. Control mice were All the animals that died or that were sacrificed when given tap water without diabenol. Fresh solution was moribund were autopsied. At autopsy their skin and prepared ex tempore three times a week.
all internal organs were examined. Revealed neopla- I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 sia were classified according to the recommendations Parameter α is often characterized by the value of of the International Agency of Research on Cancer mortality rate doubling time (MRDT), calculated as (IARC) as "fatal" (i.e., those, that directly caused the ln(2)/α. Parameters for the model were estimated from death of the animal) or as "incidental" (for the cases empirical data by use of the maximum likelihood in which the animal died of a different cause) ( method implemented in the Gauss statistical system (Confidence intervals for the mors, as well as the tissues and organs with suspected aging rate parameter estimates were calculated by tumor development, were excised and fixed in 10% profiling the log-likelihood function neutral formalin. In rats, intestines were opened lon- gitudinally. The position and size of each tumor wererecorded on special charts (Afterroutine histological processing, the tissues were em- 3. Results
bedded in paraffin. Thin, 5–7 m histological sectionswere stained with hematoxylin–eosine and were micro- 3.1. Experiment with female NMRI mice scopically examined; regarding the experimental groupto which the mice belonged this was a blind process.
3.1.1. Age-related body weight dynamics Tumors were classified in accordance with IARC rec- The body weight gain dynamics was studied in the control and treated with diabenol groups of mice. Itwas shown that the body weight of the mice in both 2.5. Statistics groups increased with age, exceeding by 12 monthsthe body weight of 3-month-old animals by 17.6% Experimental results were statistically processed by in the control group, and by 18.6% in the group the methods of variation statistics (The given diabenol. There were no differences in the mean significance of the discrepancies was defined accord- body weight of mice exposed and non-exposed to the ing to Student's t-criterion, Fischer's exact method, drug during the all period of observation (data not a chi-square analysis, and a non-parametric crite- rion of Wilcoxon–Mann–Whitney (For discrepancies in neoplasm incidence to be esti- 3.1.2. Age-related dynamics of food and water mated, an IARC method of combined contingency tables calculated individually for the fatal and inci- Regular measurements have shown that the amount dental tumors as well as a preva- of food and water daily consumed by the mice in lence analysis () were the control group and treated with diabenol group were practically the same during the all period of For survival and risk analysis, Cox's method ( observation and varied from 4.9 ± 0.40 to 5.9 ± was used; for testing two groups survival 0.21 g/mouse of food in control group and from 4.8 equality, Taron's life table test (was used.
± 0.51 to 5.7 ± 0.17 g/mouse of food in diabenol- All reported values for survival tests are two sided.
treated mice whereas water consumption varied from4.3 ± 0.33 to 5.3 ± 0.23 ml/mouse in the controls and 2.6. Mathematical models and estimations from 4.0 ± 0.36 to 5.2 ± 0.43 ml/mouse in mice givendiabenol.
The mathematical model was used to describe sur- vival under the treatment. The model is the traditional 3.1.3. Age-related dynamics of estrous function in Gompertz model with survival function Investigations of the estrous function in the animals S(x) = exp − of both groups were performed every 3 months, starting α exp (αx) − 1 when the mice were 3 months of age. The following pa- where parameters α and β are associated with the rameters of estrous function were estimated: the length aging rate and the initial mortality rate, respectively.
of the estrous cycle, the relative rate of estrous cycle I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 Table 1Effect of diabenol on age-related dynamics of estrus functional parameters in NMRI mice Rate of estrous cycles of various length (%) ∗ The difference with the controls of corresponding age in the control group is significant: p < 0.05 (Fischer's exact test).
∗∗ The difference with the controls of corresponding age in the control group is significant: p < 0.01 (Student's t-test).
# The difference from the parameter at the age of 3 months in the same group: p < 0.05.
phases (in percent); and the relative number of short (<5 3.1.4. Age-related dynamics of body temperature days) and long (>7 days) estrous cycles. The relative number of animals with regular cycles and irregular Both the control and exposed to the drug mice re- cycles (persistent estrus and anestrus) was calculated vealed significant decrease in body temperature with as well. Judging by the data presented in the age, but only at the age of 6 months, the body tem- length of estrous cycle in the control female NMRI perature was decreased in diabenol-treated mice as mice was increased with the advance in age, whereas compared with the controls: 38.6 ± 0.14 and 38.1 ± was not changed with age in the group treated with 0.19 ◦C, respectively, p < 0.05.
diabenol. In control mice, the relative number of shortestrous cycles slightly decreased with age (29.6% at the 3.1.5. Survival and longevity of female NMRI mice age of 6 months and 12.5% at the age of 12 months), Survival rate dynamics in the mice treated and non- whereas in the mice exposed with diabenol, it was prac- treated with diabenol are demonstrated in As tically constant during the entire period of observation.
shown in the table, the survival rate dynamics were In the controls of the oldest age group, irregular cy- in general similar in both groups up to the age of 12 cles were registered in 57.9% of animals and only in months. However, it is worthy to note that the mortality 6.5–10% at the age of 3 or 6 months. Treatment with di- in the group given diabenol was decreased between the abenol significantly reduced a relative number of mice 12th and the 15th months of their life as compared to with irregular estrous cycles at the age of 12 months as the control group (Drastic increase in mortality compared with controls (p < 0.05).
between months 12 and 13 in control group related Thus, these data suggest that the long-term admin- mainly to death of tumor-bearing animals in this period istration of diabenol slows down age-related changes (lymphomas and metastasizing mammary carcinomas) in estrous function.
Table 2Effect of diabenol on survival distribution in female NMRI mice No. of survivors at the age (months) ∗ The difference with the corresponding age in the control group is significant: p < 0.01 (Fischer's exact test).
I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 Table 4Effect of diabenol on incidence, localization and type of tumors infemale NMRI mice Number of tumor-bearing Age of the first mammary tumor detection (days) Mean latency of mammary adenocarcinomas (days) Total number of mammary Number of mammary Number of mammary adenocarcinomas permouse Maximum tumor size (cm) Number of mice with metastases of mammaryadenocarcinoma intolungs (%) Number of mice with malignant lymphoma ∗ The difference with controls is significant: p < 0.05 (Fischer's exact test).
∗∗ The difference with controls is significant: p < 0.01 (Student's The mean and maximum life span of mice in the control group and in the group given diabenol was prac-tically the same. However, the life span in the last 10% Fig. 1. Effect of diabenol on survival and tumor yield curves in of the mice increased by the duration of diabenol treat- female NMRI mice. (A) Ordinate, number of mice (%); abscissa, ment (by +5.0%, p < 0.05) (According to the age (days); 1, control; 2, diabenol. (B) Ordinate, number of tumor- log-rank test difference between distributions of life bearing mice (%); abscissa, age (days); 1, control; 2, diabenol.
spans in the control and experimental groups is statis-tically significant with p-value = 0.0737.
Table 3Effect of diabenol on parameters of life span in female NMRI mice Mean life span (days, mean ± S.E.) Mean life span of last 10% of survivors (days) Maximum life span (days) Ageing rate, α (days−1) 0.0140 (0.0139; 0.0157) 0.0136 (0.0133; 0.0145) 49.62 (44.13; 49.85) 51.09 (47.94; 51.99) Note: Mean life span are given as mean ± standard error; 95% confidence limits are given in parentheses; MRDT, mortality rate doubling time.
∗ The difference with controls is significant: p < 0.05.
I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 Table 5Effect of diabenol on age-related dynamics of estrus functional parameters in HER-2/neu mice Rate of estrous cycles of various lengths (%) ∗ The difference with the controls of corresponding age in the control group: p < 0.05.
∗∗∗ The difference with the controls of corresponding age in the control group: p < 0.002 (Student's t-test).
3.1.6. Spontaneous tumor development in female detected whereas in the diabenol-treated group, one case of lymphoma has been observed. The treatment The total tumor incidence in the control mice was with diabenol significantly shifted to right the total tu- 50% (Mammary carcinomas and malignant mor yield curve as compared with the control group lymphomas developed most frequently, which cor- responded to the oncological characteristics of fe- Thus, it is worth noting that the treatment with di- male NMRI mice ). The abenol inhibits the development of mammary carcino- time of the first mammary tumor detection was in- mas and malignant lymphomas in NMRI mice.
creased by 3 months in the group treated with dia-benol, and the mean latent time of mammary tumors 3.2. Experiment with female transgenic was increased by 2.3 months by the drug HER-2/neu mice According to the log-rank test, difference betweendistributions of age of first mammary adenocarcino- 3.2.1. Age-related body weight dynamics mas is statistically significant with p-value = 4.15 × The body weight of the mice in both control and 10−13; difference between distributions of life spans diabenol-treated groups increased with age, exceeding of mice with fatal tumors (both mammary adenocar- by 11 months the body weight of 3-month-old animals cinoma and lymphoma) is statistically significant with by 77.3% in the control group, and by 73.2% in the p-value = 0.182. The incidence of lung metastases of group given diabenol. There were no differences in the mammary carcinomas was 10% in the control group mean body weight of mice exposed and non-exposed and 0% in the group given diabenol. In the control to the drug during the all period of observation (data group, four cases of malignant lymphoma have been are not shown).
Table 6Effect of diabenol on parameters of life span in female HER-2/neu mice Mean life span (days, mean ± S.E.) Mean life span of last 10% of survivors (days) Maximum life span (days) Aging rate, α (days−1) 0.0307 (0.0281; 0.0337) 0.0374 (0.0351; 0.0415) 22.58 (20.58; 24.66) Note: Mean life spans are given as mean ± standard error; 95% confidence limits are given in parentheses; MRDT, mortality rate doubling time.
∗ The difference with controls is significant: p < 0.05.
I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 3.2.2. Age-related dynamics of food and waterconsumption The amount of food and water daily consumed by the mice in the control group and treated with diabenolgroup were practically the same during the all periodof observation and were similar in both groups andvaried from 3.27 ± 0.48 to 4.20 ± 0.14 g/mouse offood in control group and from 2.63 ± 0.14 to 4.03 ±0.24 g/mouse of food in diabenol-treated mice whereaswater consumption varied from 4.5 ± 0.51 to 5.85 ±0.77 ml/mouse in the controls and from 4.15 ± 0.84 to5.73 ± 0.45 ml/mouse in mice given diabenol.
3.2.3. Age-related dynamics of estrous function inmice The length of estrous cycle in the control female HER-2/neu mice was increased with the advance inage, whereas it was not changed with age in the miceexposed to the drug (In control mice, the rel-ative number of short estrous cycles slightly decreasedwith age (36.4% at the age of 5 months and 14.3% atthe age of 9 months), whereas in the mice exposed todiabenol it was practically constant during the entireperiod of observation. In the control group irregularcycles were registered in 33.3% of mice at the age of5 months and in 19.2% mice at the age of 9 months.
The exposure to diabenol significantly reduced a rela-tive number of mice with irregular estrous cycles at theage of 5 and 9 months ( Thus, these data suggest that the long-term adminis- tration of diabenol inhibits the aging of the reproductive Fig. 2. Effect of diabenol on survival and tumor yield curves in fe- male HER-2/neu mice. (A) Ordinate, number of mice (%); abscissa,age (days); 1, control; 2, diabenol. (B) Ordinate, number of tumor- 3.2.4. Age-related dynamics of body temperature bearing mice (%); abscissa, age (days); 1, control; 2, diabenol.
Both the control and exposed to the drug mice re- experimental groups are identically distributed with vealed significant decrease in body temperature with probability p = 0.923. However, maximum life span age. There was no difference in average body temper- was increased by 1 months in the group treated with ature between the both groups during the entire period diabenol as compared with the controls al- of observation (data are not shown).
though aging rate parameter α was slightly higher indiabenol-treated mice.
3.2.5. Survival and longevity of female HER-2/neumice 3.2.6. Spontaneous tumor development in female Survival dynamics in the mice treated and non- HER-2/neu mice treated with diabenol are demonstrated at The The incidence of mammary adenocarcinomas in the survival dynamics were in general similar in both control female HER-2/neu mice was 100%. The treat- groups during the all period of observation. Accord- ment with diabenol failed influence significantly any ing to the log-rank test life spans in the control and parameter of carcinogenesis in this strain of mice. Nev- I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 ertheless, some parameters of spontaneous carcinogen- esis have a tendency to decrease in the group treated by Effect of diabenol on 1,2-dimethylhydrazine (DMH)-induced colontumorigenesis in rats diabenol: total incidence of tumors (100% and 92%);total number of mammary adenocarcinoma (195 and 179), number of mice with metastases of mammary adenocarcinoma in the lung (48.3% and 35.7%), max- Number of tumor-bearing imum size of the metastases (0.52 ± 0.11 and 0.46 Colon (all parts) 0.08 cm), in control and diabenol-treated group, re- No. of tumor-bearing spectively. The treatment with diabenol also slightly shifted to the right the tumor yield curve as compared with the control group ( No. of tumors per rat Thus, the treatment with diabenol revealed only In tumor-bearing rats slight tendency to the inhibition of the developmentof mammary carcinomas in HER-2/neu mice.
Mean tumor size (mm2) 3.3. Effect of diabenol on No. of tumor-bearing carcinogenesis in rats No. of tumors per rat The treatment with diabenol failed influence the In tumor-bearing rats body weight gain, water and food consumption in rats Mean tumor size (mm2) exposed to DMH as compared to those treated with thecarcinogen alone (data are not shown). At the end of ex- periment, intestinal tumors were found in the majority No. of tumor-bearing Macroscopically, these neoplasms were exophytic No. of tumors per rat or endophytic. Several cases of ulcerative-infiltrative forms were observed as well. Microscopically, differ- In tumor-bearing rats ent types of malignant intestinal tumors were found, Mean tumor size (mm2) predominantly, tubular adenocarcinomas. Ca in situ,superficial carcinomas, mucinous and signet-ring car- cinomas were also registered. All these types of car- No. of tumor-bearing cinomas are typical for neoplasms induced by DMH No. of tumors per rat The data on the effect of diabenol on the develop- ment of colon tumors induced by DMH are presented in In tumor-bearing rats otal incidence of intestinal tumors was Mean tumor size (mm2) similar in both groups. However, the multiplicity (meannumber of tumors per rat in a group) in animals treated ∗ The difference from the controls is significant: p < 0.05.
with diabenol was decreased by 30% as compared with The difference from the controls is significant: p < 0.01.
∗∗∗ The difference from the controls is significant: p < 0.002.
the parameter in rats exposed to DMH alone. Most ex-pressed inhibiting effect of diabenol was revealed inascending colon. The incidence of tumors of this site control animals (DMH alone), the number of rats with was decreased more than two times as compared to six and more tumors per animal was 2.1 times higher the control group, and the multiplicity of tumors was than in the group treated with the carcinogen and dia- decreased by three times.
benol (42.1% and 20%, correspondingly). Analysis of The claster analysis of distribution of animals with tumor size distribution has shown that in the descend- different numbers of colon tumors has shown that in ing colon of animals from the control group small tu- I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 icantly inhibited colon carcinogenesis. These observa- Effect of diabenol on a distribution of colon tumors by growth pattern, tions are in agreement with the data obtained with an- differentiation rate and invasion depth (in percent to total number of tidiabetic biguanides phenformin, buformin and met- formin. It was shown in earlier studies that phenformin and buformin increase the life span and inhibits sponta- neous carcinogenesis in female C3H/Sn mice ( female outbred rats ( inhibits colon carcinogenesis induced by DMH Differentiation rate well as carcinogenesis induced by some other chemical carcinogens and ionizing radiation ( ∗ The significance from the control (DMH) is significant: p < 0.001 ment with metformin prolongs life span of rats (G.
(Fischer's exact test).
Roth, personal communication) and mice ( ∗∗ The significance from the control (DMH) is significant: p < 0.05.
and inhibits pancreatic carcinogenesis in ham-sters ( mors (<51 mm2) appeared less frequently in compari- Like the biguanides, diabenol slows down the age- son with the group given diabenol (58.7% and 71.4%, related disturbanses in the estrus function of rodents. It is worthy of note that metformin improves menstrual The results of morphological analysis (ave regularity, leading to spontaneous ovulation, and en- shown that the tumors with exophitic pattern of growth hanced the induction of ovulation with clomiphene more frequently developed in diabenol-treated rats in citrate in women with polycystic ovary syndrome comparison with the controls. Opposite situation was with endophytic colon tumors. Tumors in the group treated with diabenol more frequently were highly dif- The use of phenformin in humans has been limited ferentiated and less invasive as compared with the con- the last two decades because of an association with lac- trol group. Thus, these data show the inhibitory effect tic acidosis. Metformin does not increase risk for lactic of diabenol on DMH-induced colon carcinogenesis.
acidosis or increase lactate levels in type 2 diabetes(but has some adverse effects, includingrenal insufficiency ( and gastrointestinal side effects ( Our experiments have shown that the long-term Diabenol is not biguanide and belongs to deriva- treatment with the antidiabetic drug diabenol slowed tives of benzoimidazole. It seems that this drug could down age-related disturbances in estrous function and be free of adverse effects typical for biguanides. It was increased life span of all and 10% most long-living shown that diabenol decreases aggregation of throm- female NMRI mice. The treatment with diabenol in- bocytes and erhythrocytes as well as blood viscosity hibited spontaneous tumor incidence and increased the in diabetic animals (This mammary tumor latency in these mice. In short-living effect gives an additional advantage to use of diabenol transgenic HER-2/neu mice, the drug also slowed down as a geroprotector.
age-related changes in estrous function in HER-2/neu Several years ago, it was firstly suggested to use mice, slightly inhibited tumrigenesis but failed influ- biguanide antidiabetics as mimetics of CR and a poten- ence survival of these mice. In rats exposed to 1,2- tial anti-aging treatment ( dimethylhydrazine, the treatment with diabenol signif- Although it is known that free radicals are produced I.G. Popovich et al. / The International Journal of Biochemistry & Cell Biology 37 (2005) 1117–1129 during metabolic reactions, it is largely unknown which Recently, it was shown that the expres- factor(s), of physiological or pathophysiological signif- sion of mammalian Sir2 (SIRT1) is induced in CR icance, modulate their production in vivo. It has been rats as well as in human cells that were treated with suggested that hyperinsulinemia may increase free rad- serum from these animals (Long- icals, and therefore, promote aging, independent of lived mutant mice and CR rodents are protected from cancer despite attenuating apoptosis possibly because Plasma levels of lipid hydroperoxides are higher, and their cells possess increased defences and repair mech- antioxidant vitamins are lower in individuals who are anism and they retain the ability to undergo apoptosis resistant to insulin-stimulated glucose disposal but oth- if the damage is beyond repair ( erwise glucose tolerant, nonobese, and normotensive It was observed that phenformin inhibits proliferation (This finding indicates that en- and induced enhanced and transient expression of the hanced oxidative stress is present before diabetes en- cell cycle inhibitor p21 and apoptosis in human tumor sues, and therefore, cannot simply be explained by cells lines (The possibility that di- overt hyperglycemia. There is substantial evidence abenol stimulate an active defence response in the ani- supporting the hypothesis that selective resistance to mal can not to be excluded and needs an experimental insulin-stimulated (muscle) glucose disposal and the testing. Thus, the results of our experiments together consequential compensatory hyperinsulinemia trigger with the data and findings discussed above provide ev- a variety of metabolic effects, likely resulting in ac- idence that diabenol is promising geroprotector and celerated oxidative stress and aging ( The anti-diabetics biguanides inhibit fatty acid oxidation, inhibit gluconeogenesis in the liver, in- crease the availability of insulin receptors, inhibitmonoamine oxidase (increase sensi- Alexandrov, V. A., Anisimov, V. N., Belous, N. M., Vasilyeva, I. A., & tivity of hypothalamo–pituitary complex to negative Mazon, V. B. (1980). The inhibition of the transplacental blasto-mogenic effect of nitrosomethylurea by postnatal administration feedback inhibition, reduce excretion of glucocorti- of buformin to rats. Carcinogenesis, 1, 975–978.
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Oncological outcome of malignant colonic obstructionin the Dutch Stent-In 2 trial D. A. M. Sloothaak1, M. W. van den Berg2, M. G. W. Dijkgraaf3, P. Fockens2, P. J. Tanis1,J. E. van Hooft2 and W. A. Bemelman1 on behalf of the collaborative Dutch Stent-In study group 1Department of Surgery, 2Department of Gastroenterology and Hepatology, and 3Clinical Research Unit, Academic Medical Centre, Amsterdam,The NetherlandsCorrespondence to: Professor W. A. Bemelman, Department of Surgery, Academic Medical Centre, PO Box 22660, 1100 DD Amsterdam, The Netherlands(e-mail: [email protected])