Efecto combinado de la interacción genética sobre el desarrollo del bocio nodular con tiroiditis autoinmune y adenoma tiroideo en los habitantes de Bucovina del Norte / Combined effect of gene-gene interaction on the development of nodular goiter with autoimmune thyroiditis and thyroid adenoma in the inhabitants of Northern Bukovyna

ABSTRAC This article presents the results of a comprehensive analysis of the combined influence of genetic polymorphisms associated with various links of apoptosis regulation (BCL-2, CTLA-4 and APO-1/Fas) on the development of nodular goiter with autoimmune thyroiditis and thyroid adenoma in the studied population. The analysis was performed using the Multifactor Dimensionality Reduction (MDR) method by calculating the prediction potential. Graphic models of gene-gene interaction with the highest cross-validation consistency created by the MDR method showed complex "synergistic or independent" impact of polymorphic loci of the CTLA-4 (+49G/A), Fas (-1377G/A) and BCL-2 (63291411 A>G) genes on the onset of thyroid pathology in general, or its individual types (nodular goiter with autoimmune thyroiditis and thyroid adenoma) in the population of Northern Bukovyna.

RESUMEN Este artículo presenta los resultados de un análisis exhaustivo de la influencia combinada de polimorfismos genéticos asociados a diversos enlaces en la regulación de la apoptosis (BCL-2, CTLA-4 y APO-1/FAS) sobre el desarrollo de bocio nodular con tiroiditis autoinmune y adenoma tiroideo en la población estudiada. Para ello, se utilizó el método de reducción de dimensionalidad multifactorial (MDR) mediante el cálculo de los potenciales de predicción. Los modelos gráficos de interacción gen-gen con la mayor consistencia de validación cruzada creada por el método MDR mostraron un complejo impacto «sinérgico o independiente¼ de los loci polimórficos de los genes CTLA-4 (+49G/A), FAS (-1377G/A) y BCL-2 (63291411A>G) en el inicio de la patología tiroidea en general, o sus tipos individuales (bocio nodular con tiroiditis autoinmune y adenoma tiroideo) en la población de Bucovina septentrional.

Molecular analysis of Bcl-2 and cyclin Dl expression in differentially expressing estrogen receptor breast cancer MCF7, T47D and MDA-MB-468 cell lines treated with adriamycin

Bcl-2 and Cyclin Dl [CCND1] are key elements in cancer development and progression. Bcl-2 acts as a cell death suppressor and is involved in apoptosis regulation. Cyclin Dl is an important regulator of Gl/S phase of the cell cycle progression. In addition, estrogen receptor [ER] is an important prognostic factor in breast cancer cells. Therefore it is important to determine the Bcl-2 and CCND1 expression in MCF7, T47D and MDA-MB-468 breast cancer cell lines with different ER status following Adriamycin [ADR] treatment. Cytotoxicity of ADR [250 and 500nM] after 1-5 days exposure of the cell lines was evaluated by MTT assay. The mRNA and protein levels of Bcl-2 and cyclin Dl in tested cell lines were also analyzed by RT-PCR and immunocytochemistry [ICC] methods ADR cytotoxicity was highest in MDA-MB-468 and lowest in MCF7 cells in a time-dependent manner. Bcl-2 mRNA increased in MCF7 and decreased in MDA-MB-468 after exposure to ADR but it was less detectable in T47D cells. The expression of CCND1 in MCF7 with high level of ER expression was higher than the other two cell lines in untreated conditions. However, CCND1 mRNA did not show significant changes after ADR treatment. Immunocytochemical analysis did not show significant differences between Bcl-2 protein expression in the presence or absence of ADR in MDA-MB-468 cell line while in T47D and MCF7 cells its expression decreased after exposure to ADR. In addition to nuclear expression of cyclin Dl in all cell lines, strong cytoplasmic expression of cyclin Dl protein was observed only in MCF7 and T47D cells. The tested cell lines with different levels of ER expression showed differential molecular responses to ADR that is important in tumor-targeted cancer therapy

Evaluation of expression of B-cell lymphoma-2 of peripheral blood T lymphocytes and soluble Fas in children with allergic rhinitis and allergic rhinitis combined with asthma

The function of apoptosis [programmed cell death; PCD] is to eliminate unnecessary or dangerous cells. The balance between production and death is important in the control of cell numbers within physiological ranges. The dysregulation ofapoptosis and the expression of apoptosis-related molecules of allergen-reactive T lymphocytes have been suggested to play a key role in the development and maintenance of the inflammatory reactions in allergic diseases. The present study was undertaken to investigate the expression of apoptosis-related molecules Fas and B-cell lymphoma [Bcl]-2 of peripheral blood T lymphocytes in allergic rhinitis children and in allergic rhinitis children combined with asthma. The plasma concentration of serum Fas was measured using the enzyme-linked immunosorbent assay [ELISA]. The expression of the Bcl-2 molecule in T lymphocytes was assessed by flow cytometry. We examined two groups of patients with allergy. The first group consisted of 23 children with allergic rhinitis only. The second group consists of 22 children with allergic rhinitis combined with asthma. The study was done during the season of allergy from March 2007 to September 2007. The control group comprised 20 apparently healthy, age and sex matched subjects. The results showed that the serum level of Fas was significantly elevated in allergic rhinitis patients combined with asthma when compared to patients suffering from allergic rhinitis only and to control subjects [P= 0.007]. The levels of Bcl-2 were significantly lowered in both groups of our study compared to control group [P<0.001]. In conclusion, the lower serum levels of soluble Fas in patients with allergic rhinitis in contrast to its elevation in allergic rhinitis patients combined with bronchial asthma suggesting a different pathogenesis for allergic rhinitis and bronchial asthma at the apoptosis-linked step. While the lower levels of Bcl-2 in both groups revealed that, there was need for further investigations on the dysregula-tion of activation-induced cell death of T lymphocytes

Gene rearrangements in follicular lymphoma among Indonesian.

AIM: Gene rearrangement has an important role in the management of lymphoma. We investigated the rearrangements of B-cell leukaemia/lymphoma 2 (BCL2), BCL6 and Paired homeobox 5 (PAX5) genes in Indonesian follicular lymphoma (FL) patients. METHODS: We examined gene rearrangements using various kinds of polymerase chain reactions (PCRs) on 24 patients' peripheral blood DNA. RESULTS: BCL2 rearrangement was found in 58% (14 of 24 patients), 8 at mbr (major breakpoint region), 2 at mcr (minor cluster region) and 4 at icr (intermediate cluster region), respectively. No rearrangement in BCL6 and PAX5 was detected. There was a significant difference in the incidence of spleen involvement between patients with BCL-2 rearrangement and without it (50% vs. 11%, p=0.04). BCL-2 rearrangement was correlated with spleen involvement (OR=9) and anemia (OR=2.3). CONCLUSION: BCL2 rearrangement in Indonesian FL was higher than previous reports from other Asia countries (58% vs. 48%, respectively). Our method using peripheral blood DNA might be useful for the molecular diagnosis of FL.

Prognostic significance of Bcl-2, Bax and p53 expression in breast cancer

Aim: of this study was to investigate the significance of apoptosis-related genes bcl-2 and Bax in breast carcinoma cases treated with radical surgery plus radiotherapy and adjuvant chemotherapy for at least 1 year and their relation with expression of p53

Methods: After surgical resection imunohistochemistry was performed to determine Bcl-2, Bax, p53 and estrogen receptor [ER] expression in paraffin-embedded tissues of 50 invasive breast cancers. And overall survival was assessed

Results: Bcl-2, Bax, P53 and ER immunostaining displayed a positive relation with increasing histologic grade [P=0.000] and negative relation with time staging. Bcl-2 displayed a negative relation with p53 [P=0.035] and a positive relation with Bax and ER [P= 0.003 and P= 0.011 respectively]. Expression of Bc12 was associated significant improvement in overall survival [P=0.01]

Conclusion: regulation of apoptosis is important in invasive ductal carcinoma. These results indicate that bcl-2 expression is significantly associated with hormonal receptor status so it is good prognostic marker, and that p53 is a significant prognostic marker. No significant relation between Bax and overall survival in relation to the stage, p53 or ER

Bcl-2 gene rearrangement in Jordanian follicular and diffuse large B-cell lymphomas

Follicular lymphoma [FL], a common subtype of non-Hodgkin's lymphoma [NHL] in the West, represents a rare subtype in Jordan. Bcl-2 gene rearrangement plays a crucial role in the biology of the vast majority of FL and a substantial number of diffuse large B-cell lymphoma [DLBCL] in the West; but its presence has not been studied in Jordan. Our aims are to document if bcl-2 gene rearrangement exists in Jordanian FL and DLBCL, and if present to determine whether its frequency among these lymphomas is different from the West and therefore may be responsible for some of the epidemiological differences seen between Jordan and the West. The study was conducted in the year 2001 using polymerase chain reaction [PCR], to detect bcl-2 gene rearrangement in paraffin sections in 5 FL and 23 DLBCL cases diagnosed at the Department of Pathology at Jordan University of Science and Technology, Irbid, Jordan. Two sets of primers including the major breakpoint region [MBR] and the minor cluster region [MCR] were used. Amplifiable DNA was extracted from all cases. Bcl-2 gene rearrangement was seen among 4 [80%] of 5 FL cases, and 8 [35%] of 23 DLBCL cases. The majority of the rearrangements involved the MBR; however, one fourth of cases [one of 4 FL; 2 of 8 DLBCL] with bcl-2 rearrangement involved the MCR. Bcl-2 gene rearrangement was seen in the vast majority of Jordanian FL cases and approximately one third of all DLBCL cases. These figures are similar to those reported in the West, and therefore bcl-2 gene rearrangement does not help in explaining the epidemiological differences of NHL between Jordan and the West. The presence of bcl-2 gene rearrangement in DLBCL may define a subset of lymphomas that may be biologically and clinically unique and different from the rest of DLBCL

Expression and functional activity of receptor-Ck in mononuclear cells of a homozygous hypercholesterolemic family.

The study was addressed to explore the expression and functional activity of a novel cholesterol-specific cell surface receptor-Ck in a typical homozygous familial hypercholesterolemic family. Functional activity of receptor-Ck was characterized by its ability to downregulate Bcl-2 gene expression through a 47 kDa factor having an affinity for the sterol-regulatory element in the promoter region of this gene. The result of such a study revealed normal expression and functional activity of receptor-Ck accompanied by a lack of Apolipoprotein B-specific low-density lipoprotein receptor gene expression in the mononuclear cells derived from these patients. On the basis of these results, it is tempting to speculate that receptor-Ck may be involved in the maintenance of cellular cholesterol homeostasis observed in homozygous familial hypercholesterolemic patients.

The experimental and clinical study on the effect of curcumin on cell cycle proteins and regulating proteins of apoptosis in acute myelogenous leukemia / 华中科技大学学报（医学）（英德文版）

To investigate whether the Bcl-2 gene family is involved in modulating mechanism of apoptosis and change of cell cycle protein induced by curcumin in acute myeloid leukemia HL-60 cell line and primary acute myelogenous leukemic cells, the Bcl-2 family member Mcl-1, Bax and Bak and cell cycle proteins including P27kipl, P21wafl, cyclin D3 and pRbp- were selected and their expression detected by SABC immuno-histochemical stain method. The attitude of sub-G1 peak in DNA histogram was determined by FCM. The TUNEL positive cell percentage was identified by terminal deoxynucleotidyl transferase (TdT)-mediated Biotin dUNP end labeling technique. It was found that when HL-60 cells were treated with 25 mumol/L curcumin for 24 h, the expression level of Mcl-1 was down-regulated, but that of Bax and Bak up-regulated time-dependently. There was significant difference in the expression level of Mcl-1, Bax and Bak between the curcumin-treated groups and control group (P < 0.05-0.01). At the same time, curcumin had no effect on progress of cell cycle in primaty acute myelogenous leukemia at newly diagnosis, but could increase the peak of Sub-G1 (P < 0.05), and down-regulate the expression of Mcl-1 and up-regulate the expression of Bax and Bak with the difference being statistically significant. The expression of P27kipl, P21wafl and pRbp- were elevated and that of cyclin D3 decreased in the presence of curcumin. These findings suggested that the Bcl-2 gene family indeed participated in the regulatory process of apoptosis induced by curcumin in HL-60 cells and AML cells. Curcumin can induce apoptosis of primary acute myelogenous leukemic cells and disturb cell cycle progression of HL-60 cells. The mechanism appeared to be mediated by perturbing G0/G1 phases checkpoints which associated with up-regulation of P27kipl, P21wafl and pRbp- expression, and down-regulation of cyclin D3.

Spontaneous programmed cell death of peripheral blood mononuclear cells from HIV-infected persons is decreased with interleukin-15

Interleukin 15 (IL-15) is an important regulatory cytokine in cellular immunity. In vitro replacement of IL-15 has been shown to enhance immunity in Human immunodeficiency virus type 1 (HIV-1) infected lymphocytes. We evaluated the effect of IL-15 on the survival of peripheral blood mononuclear cells of HIV patients by examining in vitro lymphocyte apoptosis, and correlated the process with Bcl-2 and Fas gene regulation. Peripheral blood mononuclear cells (PBMC) from 21 HIV-infected adults and 24 HIV-seronegative healthy individuals were isolated and cultured to determine the effect of escalating doses of IL-15 (0, 1, 10, 100, 1000 ng/mL) on apoptosis. Lymphocyte proliferation assay with (3H) TdR was measured and Bcl-2 and Fas gene regulation was observed. The results were as follows: 1) IL-15 reduced culture induced lymphocyte apoptosis in HIV patients in a dose dependent manner, and reached a plateau level at a concentration of 100 ng/ml; 2) IL-15 significantly reduced the level of apoptosis after 3 days (14%) and 5 days (15%) of culture in HIV patients, while no difference was observed in HIV (-) donors; 3) The percentage of viable cells among the total number of lymphocytes was significantly enhanced by 25% in HIV patients with IL-15; 4) Bcl-2 expression was decreased in HIV patients (53.9 +/- 12.3%) compared to HIV (-) donors (93.0 +/- 3.7%), and IL-15 increased Bcl-2 expression by 21.2 +/- 5.2% in HIV patients; 5) Fas expression was increased in HIV patients (70.2 +/- 4.6%) compared to HIV (-) donors (32.4 +/- 4.3%), and IL-15 increased Fas expression by 8.4 +/- 1.2% in HIV (-) donors. Our findings indicate that IL-15 may influence immunologic abnormalities in HIV infection, particularly its ability to prevent apoptosis of lymphocytes by suppressing the down-modulation of Bcl-2. This may provide an experimental basis for IL-15 immunotherapy.

Study on the metastatic mechanisms of human giant-cell lung carcinoma comparison between the strains C and D.

The biologic characteristics of the two human giant-cell lung carcinoma strains with high (strain D) and low metastatic potential (strain C) were studied, including karyotype of chromosome, intracellular free calcium ([Ca2+]i), morphologic changes of cell surface and the expression of nm23-H1, p53, ras, c-myc, c-erbB2, bcl-2 genes and PCNA. The correlation between different biologic features and the metastatic potential of the two strains was analyzed. We found: 1) Both strains had the karyotypic abnormality of -13, -14, -15, +20, +21 with seven same marker chromosomes. Only strain D had the karyotypic abnormality of +7, -17, -18, +X, 7p+; 2) [Ca2+]i of the strain C (984.7 +/- 573.8) and D (517.6 +/- 216.6) was significantly different (p < 0.05). The amplitude of intracellular calcium oscillations of strain C was lower than the one of strain D; 3) strain C had more villous-like protrusions on the cell surface, whereas strain D had more bubble-like protrusions; 4) The expression of nm23-H1 and p53 protein of strain C was all higher than that of strain D. The expression of PCNA of strain C was lower than strain D; 5) nm23-H1 mRNA levels of strain C was lower than that of strain D. We consider that the karyotype of chromosomes, intracellular free calcium, the structure of cell membrane and the expression of nm23-H1 gene, p53 gene, PCNA could be closely related to the metastatic potential of human giant-cell lung carcinoma. They could be used as the sign for judging whether the tumor will metastasize in clinical practice as well as in judging the prognoses of patients.