Experimental, Clinical and Morphological Analysis of H-Ras Oncoproteins for Locally Advanced Breast Cancer.

BACKGROUND: Activated forms of Ras are enhanced in both breast cancer as well as the cell lines with EGFR and HER2 expression. Therefore, H-Ras could be activated in breast tumours in the absence of direct mutational activation of Ras itself and could contribute to 20-50% of the cases. Expression inhibition, signal transduction interruption from H-Ras to the nucleus could become a promising therapeutic target. AIM: The aim of this study was to investigate the clinical and morphological criteria of locally advanced breast cancer and the expression of H-Ras oncoprotein in patients who have been subjected to different regimens of farnesyltransferase inhibitor. METHODS: H-Ras status was assessed by immunohistochemistry (IHC). RESULTS: An association between the expressions of H-Ras and Her2/neu (p = 0.001) as well as the tumour proliferation index Ki-67 (p = 0.001) in patients with breast cancer was established. Analysis of the relationship between H-Ras expression showed a relatively strong association with progression-free survival both before the treatment (V = 0.47; p = 0.001) and after the treatment (V = 0.45; p = 0.001). These results may indicate the clinical applicability of H-Ras as a prognostic factor or serve as a therapeutic target for breast cancer treatment. CONCLUSION: These results could indicate the potential clinical application of H-Ras as a prognostic factor or a therapeutic target for breast cancer treatment.

LONG-TERM RESULTS OF COMBINATION THERAPY FOR LOCALLY ADVANCED BREAST CANCER.

Breast cancer ranks first among the malignant tumors in women. In locally advanced breast cancer (LABC) treatment starts with neoadjuvant chemotherapy (CTx) with the standard regimens CMF, FAC, АС. A cytostatic drug, Arglabin, isolated and produced from Artemisia glabella Kar. et Kir., an endemic plant growing in Central Kazakhstan, has been under investigation in clinical trials. The research is aimed at investigating the long-term results of combination therapy of locally advanced breast cancer including different chemotherapy regimens and Arglabin as monotherapy. The present research includes 93 patients diagnosed with LABC aged from 35 to 75 years, including 60 patients with Stage 2, and 33 patients with Stage 3 breast cancer. All patients were split into 3 groups, two experimental and one control group. The control group consisted of 36 patients with LABC who underwent 4 cycles of neoadjuvant chemotherapy according to AC regimen (doxorubicine 50 mg/m2, cyclophosphamide 500 mg/m2). The experimental group 1 consisted of 30 patients who received 4 cycles of chemotherapy according to AC+Arglabin regimen (Arglabin 370 mg/m2 within 7 days), while experimental group 2 consisted of 27 patients who underwent 4 cycles of Arglabin as monotherapy. Actuarial calculations of the overall survival (OS) and disease-free survival (DFS) rates were done according to the Kaplan-Meier method, while the differences in indicators of control and experimental groups were estimated using the following methods: Cox's F-Test, χ2, Gehan's Wilcoxon Test. Overall one- and two-year survival in all groups of patients was 100%. Three-year survival rate in patients treated with chemotherapy according to AC regimen was (40,0±8,2)%, in patients combining AC chemotherapy with Arglabin it was (60,0±8,9)%. The lowest three-year survival rate (28,0±8,6)% was observed in patients treated with Arglabin as a monotherapy. The three-year survival rate in patients with breast cancer is statistically insignificant (χ2=4,407 at p=0,11042) between all groups; however, at the paired comparison by Gehan's Wilcoxon criterion a statistically significant difference has been observed between the group treated with Arglabin as monotherapy, and the group receiving chemotherapy according to AC+Arglabin regimen. The highest disease-free survival rates have been observed in the group of patients receiving chemotherapy according to AC+Arglabin regimen: 1-2-3-year survival rates are 100%, 100%, and 58%, respectively. In the group of patients who underwent chemotherapy according to AC regimen, 1-2-3-year disease-free survival rates are 92%, 92%, and 30%, respectively. Arglabin included in AC regimen positively increases a 3-year disease-free survival rate by 28% as compared to the standard regimen.

[Mother thyroid status and development of enzymes of initial and final phases of carbohydrate hydrolysis]. / Aktivnost' tireoidnogo statusa materi na razvitie fermentov nachal'nogo i zakliuchitel'nogo étapov gidroliza uglerodov.

The daily treatment of a lactating mother with thyroxine 50 micrograms/100 g b.w. from the first to 21st day of lactation resulted in an increase of the thyroxine level in serum and earlier induction of alpha-glycosidases (pancreatic alpha-amylase and enteral maltase, saccharase) and in the earlier repression of beta-galactosidases (intestinal lactase) in rats. In contrast, sucklings fed by hypothyroid mothers showed a low level of thyroxine and a delayed induction of alpha-glucosidases but ameliorated the repression of lactase.

Delay in natural decline of lactase activity in the small intestine of prematurely weaned rats as related to changes in their thyroid status.

Premature weaning of suckling rats on the 15th day of life delayed the natural reduction of lactase activity in the small intestine and reduced the level of T4 in serum. Preliminary treatments (from day 12 to 14 of life) with exogenous T4 (1 mg/kg BW) or exposure to cold (10 degrees C) significantly increased the level of T4 in serum on the 15th day of life, and sharply accelerated the reduction of lactase activity. It is concluded that the delay in lactase reduction in prematurely weaned rats may be related to a decrease in serum thyroxine level. Cold as well as exogenous thyroxine may be considered as a means for regulation of the functional maturation of the small intestine.

[The effect of insufficient nutrition during the suckling of rats on their growth and digestion]. / Vliianie nedostatochnogo pitaniia v period molochnogo vskarmlivanie krys na ikh rost i pishchevarenie.

The average weight of the bodies and the small intestine brush was 30% less in the malnourished 21-day old rats as compared with the control. The weight of the pancreas was 18% less. A decrease in the hydrolytic strength of the enzyme pools followed the changes in enzymatic activity during the primary and the last hydrolysis of food.

[The physiological mechanisms of the delay in the functional maturation of the small intestine in rat pups prematurely switched from milk to definitive nutrition]. / Fiziologicheskie mekhanizmy zaderzhki funktsional'nogo sozrevaniia tonkoi kishki krysiat pri prezhdevremennom perekhode ot molochnogo na definitivnoe pitanie.

Weaving of 15-day rats delays the natural suppression of the small intestine lactase, induces sucrase activity and decreases the thyroxine level in the blood serum. Administration of thyroxine normalizes the level of this hormone in the blood serum as well the beginning of suppression of the lactase activity.

[The effect of heat exposure on the circadian rhythm of the enzymatic activity in different sections of the rat small intestine]. / Vliianie teplovoi ékspozitsii na tsirkadnuiu ritmiku fermentativnoi aktivnosti razlichnykh otdelov tonkoi kishki krys.

Two-hour heat exposure (40-41 degrees C) was shown to change the circadian rhythms of enteral gamma-amylase, maltase and sucrase activities. The character and obviousness of the changes depended on the type of enzyme activity and the portion of the small intestine. The heat exposure did not change daily mean levels of enzymatic activities and their distribution along the small intestine.

[Environmental factors and the functional development of the digestive system]. / Faktory vneshnei sredy i funktsional'noe razvitie pishchevaritel'noi sistemy.

A single action of cold, heat and malnutrition prompts an early transition of the digestive-transport system from juvenile to adult type. Repeated actions of these factors decrease the hydrolytic-transporting function of the digestive tract, the decrease being age-dependent.

[The effect of protein-calorie food deficiency on the activity of enteral enzymes in growing rats]. / Vliianie belkovoi i énergeticheskoi nedostatochnosti pitaniia na aktivnost' énteral'nykh fermentov u rastushchikh krys.

30-day changes in enteral enzymes were followed up in test and control growing white rats. Test animals received protein- and calorie-deficient feeding. It was established that retardation of growth was more marked in protein than in calorie deficiency.

[Changes in the saccharase and lactase activities of the small intestine in suckling rats prematurely weaned from the nursing dam]. / Izmeneniia sakharaznoi i laktaznoi aktivnostei tonkoi kishki krys-sosunkov, prezhdevremenno otniatykh ot kormiashchei materi.

In suckling rats prematurely weaned on the 15th, 17th, 19th or 21st day of life, the body weight and the weight of small intestine were studied as well as the activity of enteral sucrase and lactase. A delay in the gain of body weight and small intestine weight was the greater the earlier the sucklings had been weaned. The sucrase activity did not depend on the term of weaning. Whereas weaning on the 15th or 17th days of life considerably delayed the decrease in lactase activity. The latter seems to be due to the changes in the suckling's thyroid state because of thyroid depression under these conditions and absence of thyroid hormones intake from maternal milk.

[The effect of stress on the enzyme systems of the enterocyte microvilli in rat pups and the possible mechanism of the transmission of stress effects from dam to progeny]. / Vliianie stressa na fermentnye sistemy mikrovorsinok énterotsitov u krysiat i vozmozhnyi mekhanizm peredachi stressornykh éffektov ot materi k potomstvu.

In suckling rats whose mothers had had an immobilisation stress or been injected HC and T4 at the last day of their pregnancy, the sucrase induction decreased in the small intestine, if the HC was injected on the 10th day of life. The hormone injection to 1-day suckling rats did not affect the induction. These effects seem to be exerted not by the direct permeation of hormone from mother to litter but to be mediated rather by a special factor "stressin". The latter is transferred from mother to litter and decreases the sensitivity of intestinal enzymes to exogenous HC, thus exerting an antistressor effect.

[The effect of malnutrition during the suckling period on the functional development of the pancreas and small intestine]. / Vliianie nedoedaniia v period molochnogo vskarmlivaniia na funktsional'noe razvitie podzheludochnoi zhelezy i tonkoi kishki.

From the 1st until the 21st day of life, 8 sucklings were fed by 1 lactating rat (unlimited feeding) and 16 sucklings were also fed by 1 lactating rat (limited feeding). The limitation of feeding during suckling period was found to induce a growth retardation, a hypertrophy of the pancreas and small intestine, as well as a stable decrease in the activity of pancreatic and enteral enzymes.

[The effect of hyperthermia during pregnancy in rats on the postnatal ontogeny of the mechanisms of intestinal carbohydrate hydrolysis and transport in the progeny]. / Vliianie gipertermii vo vremia beremennosti krys na postnatal'nyi ontogenez mekhanizmov kishechnogo gidroliza i transporta uglevodov u potomstva.

In experiments on Wistar albino rats, it has been shown that exposition of animals to a temperature 40-41 degrees C from the first to the last day of gestation selectively inhibits intestinal transport of glucose in offsprings. At the same time, membrane hydrolysis of lactose and the absorption of the produced glucose increase, maltose utilization remaining essentially unaffected.

[The formation of the enzyme systems of the initial and final stages of protein hydrolysis in rat postnatal ontogeny]. / Formirovanie fermentnykh sistem nachal'nogo i zakliuchitel'nogo étapov gidroliza belkov v postnatal'nom ontogeneze krys.

Proteolytic activity of the stomach and pancreas as well as glycyl-L-leucin-dipeptide-hydrolase activity of the small intestine after introduction of hydrocortisone on the 1-, 5-, 10-, 16- and 30th days of life was studied in rats. The enzyme system sensitivity responsible for protein hydrolysis was shown to be age-dependent on the hormone administration. The peculiarities of the stomach, pancreas and small intestine responses to the hormone administration was found: the enzyme system inducibility of the pancreas was significantly higher than that of the stomach and the small intestine during the whole suckling period.

[Age-related changes in the dipeptide hydrolase activity of different sections of the small intestine in rats in response to hydrocortisone administration]. / Vozrastzavisimye izmeneniia dipeptidgidrolaznoi aktivnosti razlichnykh otdelov tonkoi kishki krys v otvet na vvedenie gidrokortizona.

The glycyl-L-leucinedipeptidehydrolase activity in different portions of the small intestine after hydrocortisone administration on the 1st, 5th, 10th, 16th, 30th day of life was studied in rats. The enzyme was mainly localized in distal portion of the small intestine for the first two weeks. The maximal reactivity of the enzyme-synthetizing system was shown after a single hydrocortisone injection during this period in proximal portion. The enzyme distribution along the small intestine became even in the end of the experimental period. The response to the hormone administration was practically absent from the 16th day of suckling life.

[The isolation of mitochondria fron different sections of the small intestine and their functional characteristics]. / Vydelenie mitokhondrii iz raznykh otdelov tonkoi kishki i nekotorye osobennosti ikh funktsionirovaniia.

High level of oxygen consumption is specific for the proximal part of small intestine in rats, particularly when mitochondria are active and when oxidative phosphorylation is uncoupled by 2.4 Dinitrophenol. High degree of coupling seems to exist in this part of the intestine. The intensity of substrate oxidation, activity of multienzyme systems of respiratory chains, and the ATP formation velocity are considerably lower in the distal part of the small intestine than in the proximal and medial ones.

[The effect of malnutrition during milk feeding on the enzyme activity of the initial and final stages of carbohydrate hydrolysis in the small intestine of growing rats]. / Vliianie nedoedaniia v period molochnogo vskarmlivaniia na aktivnost' fermentov nachal'nogo i zakliuchitel'nogo étapov gidroliza uglevodov v tonkoi kishke rastushchikh krys.

Malnutrition in the period from 10 to 17 days of postnatal life causes in young rats an early increase of intestinal alpha-glucosidase activity. However, this rise in the enzyme activity is soon changed by its decrease during normal nutrition. The decreased level of the enzyme activity remains stable during 2-3 weeks after the reduction of normal nutrition.