Investigating the Expression Levels of Bax and Bcl-2 Genes in Peripheral Blood Lymphocytes of Industrial Radiation Workers in the Asaluyeh Region.

Background: Industrial radiography uses gamma or X-ray radionuclide sources to investigate the safety of industrial materials. Industrial radiation workers receive the highest occupational radiation doses. Objective: The present study investigates the relationship between Bax and Bcl-2 gene expression variables in industrial radiation workers. Material and Methods: In this case-control study, data was collected using blood sampling from 40 workers, including two groups of non-radiation and radiation workers employed at the location. Expression levels of Bax and Bcl-2 genes were assessed in the laboratory. The environmental and absorbed doses of workers were measured using environmental and pen dosimeters. Results: Statistical analysis showed that the radiation group's Bcl-2 gene expression level was significantly higher. Findings also demonstrated a correlation between Bcl-2 gene expression and the number of workdays. Also, the Bax gene expression did not show a significant change, and the expression ratio of Bax/Bcl-2 was insignificant in the two groups. Conclusion: Exposure to low doses of radiation could promote an adaptive response in cells by increasing Bcl-2 gene expression.

Exploring the DNA2-PNA heterotriplex formation in targeting the Bcl-2 gene promoter: A structural insight by physico-chemical and microsecond-scale MD investigation.

Peptide Nucleic Acids (PNAs) represent a promising tool for gene modulation in anticancer treatment. The uncharged peptidyl backbone and the resistance to chemical and enzymatic degradation make PNAs highly advantageous to form stable hybrid complexes with complementary DNA and RNA strands, providing higher stability than the corresponding natural analogues. Our and other groups' research has successfully shown that tailored PNA sequences can effectively downregulate the expression of human oncogenes using antigene, antisense, or anti-miRNA approaches. Specifically, we identified a seven bases-long PNA sequence, complementary to the longer loop of the main G-quadruplex structure formed by the bcl2midG4 promoter sequence, capable of downregulating the expression of the antiapoptotic Bcl-2 protein and enhancing the anticancer activity of an oncolytic adenovirus. Here, we extended the length of the PNA probe with the aim of including the double-stranded Bcl-2 promoter among the targets of the PNA probe. Our investigation primarily focused on the structural aspects of the resulting DNA2-PNA heterotriplex that were determined by employing conventional and accelerated microsecond-scale molecular dynamics simulations and chemical-physical analysis. Additionally, we conducted preliminary biological experiments using cytotoxicity assays on human A549 and MDA-MB-436 adenocarcinoma cell lines, employing the oncolytic adenovirus delivery strategy.

The alteration of apoptosis-related genes in female pelvic supportive tissues with regard to menopausal status.

PURPOSE: We aimed to compare the expression levels of anti-apoptotic and proapoptotic genes in the parametrium, sacrouterine and round ligaments with respect to menopausal status in women presenting without any indication of pelvic organ prolapse (POP). We hypothesized that apoptosis related gene expressions in female pelvic tissues may be altered during menopause. METHODS: The study groups consisted of pre-menopausal (n = 10) and menopausal (n = 10) females who did not have POP symptoms. Three different types of tissue samples (Parametrium, Round Ligament and Sacrouterine Ligament) were obtained and RNA was isolated from these tissues. After purifying and quantifying RNA samples, qPCR was used to determine the expression levels of anti-apoptotic and pro-apoptotic genes. RESULTS: BCL-2 gene expression levels were significantly lower in all the tissues of menopausal patients compared to those of premenopausal patients. In comparison to premenopausal patients, the sacrouterine ligament tissue BAD expression level was significantly high (p = 0.035), and the BCL-2/BAD ratio was significantly lower in menopausal patients (p = 0.006). CONCLUSION: Apoptosis-related protein levels change during menopause; pro-apoptotic gene expressions decrease and anti-apoptotic gene expressions increase. The significant alteration of BCL-2 and BAD expression in sacrouterine ligament with respect to menopausal status was observed and this suggested that when compared to other pelvic tissues, the sacrouterine ligament, which plays a crucial role for genital organs in restoring normal pelvic anatomy and providing support, could be affected more by menopause.

The Investigation of the Association Between the Bcl-2 3'-UTR rs1564483 Polymorphism and miR-296-3p in the Development of Breast and Gastric Cancers.

Background: B-cell leukemia/lymphoma 2 (Bcl-2) gene regulates carcinogenesis by inhibiting apoptosis. This study evaluated the association of Bcl-2 3'-untranslated regions (3' UTR) rs1564483 polymorphism and miR-296-3p with the development of breast and gastric cancers. Methods: A microarray analysis was performed on the Genomic Spatial Event (GSE)29431 and GSE161533 datasets for breast and gastric cancers. Blood samples were taken from 222 (111 patients and 111 controls) and 210 (84 patients and 126 controls) individuals for breast and gastric cancers, respectively. Genomic DNA was extracted from the blood samples and genotyping was performed using real-time polymerase chain reaction (RT-PCR), followed by examining the high-temperature melting curve. Statistical analysis was conducted to examine the potential correlation between the rs1564483 polymorphism and the risk of breast and gastric cancers concerning pathological characteristics. Results: The results of the microarray showed that the Bcl-2 gene was up-regulated in gastric cancer (logFC [log fold change]: 0.65, adjusted P < .05). Clinical outcome showed no notable relationship between the rs1564483 polymorphism and breast cancer risk; however, for gastric cancer, it identified a large difference between healthy controls and patients for an allelic frequency of rs1564483 (P ⩽ .001). Moreover, an assay of different models (dominant, recessive, and co-dominant) showed a significant association between the AG genotype between control and gastric cases (Pearson chi-square test, P = .046). In addition, the prevalence of the AG genotype was greater in persons under the age of 45 and in patients with H. pylori infection (P ⩽ .001). The AG genotype was not related to smoking, although the AA genotype was associated with increased cancer incidence in smokers (P ⩽ .001). Conclusions: In silico studies and calculations of the ΔG binding of micro ribonucleic acid (miRNA) hsa-miR-296-3p to the mutant and wild alleles of the rs15644833 single nucleotide polymorphism (SNP) have revealed that Bcl-2 mRNA expression in gastric cancer decreases, thus confirming the tumor suppressor role of the Bcl-2 gene.

[Methylation Status of Apoptosis Genes and Intensity of Apoptotic Death of Peripheral Blood Lymphocytes in Persons Chronically Exposed to Radiation].

Methylation of the CpG islands of gene promoter regions is the most common epigenetic modification involved in the regulation of gene expression. A number of studies have shown that ionizing radiation can cause both hyper- and hypomethylation of DNA. Aberrant methylation affects cellular processes and can lead to the development of various pathological states. In the literature, there are few studies on the methylation status of human DNA a long time after radiation exposure. Here, the methylation level of CpG islands of the promoter regions of apoptosis genes (BCL2, ATM, MDM2, CDKN1A, STAT3, and NFKB1), and also its influence on apoptosis of peripheral blood lymphocytes in chronically exposed persons were studied. Residents of the South Ural region who were chronically exposed to radiation (after discharges of radioactive wastes into the Techa river by the "Mayak Production Association" in 1949-1956) were included in the study. It was established that the proportion of individuals with hypermethylated BCL2 gene promoter among the exposed people was statistically significantly higher than in the control group. The percentage of methylation of the ATM gene promoter weakly positively correlated with dose and age characteristics. Differences in the frequency of lymphocyte apoptosis in exposed individuals with a hypo- or hypermethylated ATM gene promoter were also established. The data indicate that, in the long-term, after chronic low intensity radiation exposure at low and medium doses, epigenetic modifications of the genome occur, which are manifested as changes in methylation of promoter regions of BCL2 and ATM genes.

Evaluation of Bax and BCL 2 Genes Polymorphisms in Iraqi Women with Breast Cancer.

The present study aimed to examine the polymorphism -938C > A of BCL-2 gene and promoter -248G>A in the Bax gene, as well as their relationship with specific clinical-pathological characteristics, in patients with breast cancer. Blood samples were obtained from 70 patients who had been diagnosed with breast cancer and 34 healthy women as the control group. Polymorphic analysis was performed using the polymerase chain reaction-restriction fragment length polymorphism assay. Anthropometric data were assessed. Estrogen receptor (ER), human epidermal growth factor receptor 2 (Her-2),  and progesterone receptor (PR) were measured by immunohistochemistry. The data of age and body mass index (BMI) demonstrated no significant variations between the two groups (P>0.05). The results of HER-2 revealed that 42.86% of breast cancer patients reflected positively for Her-2/neu expression, while 24.29% reflected negative results of Her-2/neu. Moreover, the results of ER revealed that 42.86% and 28.57% of subjects were positive and negative ER, respectively; moreover, the missing data was 28.57%. In addition, the results of PR indicated that 35.71% of patients (25/70) were positive for PR, while 28.57% reflected negative results, and the missing results were 35.71%. The genotype and allele frequencies of BCL-2(-938C>A) were not statistically significant in women with breast cancer and the control group (P=0.574, P=0.533) for heterozygous and recessive models, respectively. The genotype of BCL-2(-938C>A) in control and patients in codominant, dominant, recessive, and additive models demonstrated no significant variations of all genotypes in all groups. Genotypes and allele frequencies for Bax (-248G>A) in patients with breast cancer and control indicated that the frequencies of GG, AG, and AA genotypes in cases were 16.67%, 3.33%, and 80 %, while in controls, these values were 3.23 %, 58.06 %, and 3.23 %, respectively. The heterozygous genotype (AG) in the codominant model was OR=36.00 (95% CI 4.5608 - 284.1608; P=0.0007). In comparison with the wild type (GG), there was a 36-fold increase in the risk of breast cancer. Furthermore, the findings of this study revealed a significant correlation between Bax (-248G>A) polymorphism and breast cancer risk under the dominant and overdominant (OR=6.33; 95% CI 2.2604 -17.7452; P=0.0004, and OR=40.154; 95% CI 5.1365 - 313.8949; P=0.0004, respectively. The recessive model revealed that there was a decreased risk of breast cancer (OR= 0.167; 95% CI 0.0303 to 0.9168; P=0.039). Based on the results, it can be concluded that there were no significant variations in BCL-2 (-938C>A) polymorphism of all genotypes models when breast cancer women are compared with healthy ones. In a similar vein, there was no significant association between the BCL-2 (-938C>A) polymorphism and breast cancer risk under dominant, codominant, or recessive models.

Extra copy number of BCL2 is correlated with increased BCL-2 protein expression and poor survival in diffuse large B-cell lymphoma treated with chemoimmunotherapy.

The clinical significance of extra copy (EC) genotypes of BCL2, MYC, and BCL6 have not been fully elucidated. We evaluated the EC and translocation statuses of BCL2, MYC, and BCL6 in 190 diffuse large B-cell lymphoma (DLBCL) cases using fluorescence in situ hybridization. EC genotype was sub-classified according to copy number-gained tumor cell ratio (EC1, >20% but ≤50%; EC2, >50%). Only the BCL2-EC groups, not MYC-EC or BCL6-EC groups, displayed significantly increased immunoreactivity of the corresponding protein. Moreover, the BCL2-EC2 group was significantly associated with poor overall survival (OS) and progression-free survival (PFS) in a 147 R-CHOP-treated patient subset, which was also statistically significant as per the multivariate survival analysis for PFS. No significant differences in the survival of MYC, BCL6, concurrent BCL2/MYC, BCL6/MYC, BCL2/BCL6, or triple EC groups were observed. BCL2-EC may contribute to increased BCL-2 protein expression and serve as a predictor of treatment outcomes in DLBCL.

Decoding the concealed transcriptional signature of the apoptosis-related BCL2 antagonist/killer 1 (BAK1) gene in human malignancies.

BCL2 antagonist/killer (BAK) is a multidomain pro-apoptotic effector protein, encoded by the human BAK1 gene, which has emerged as a key checkpoint in the apoptotic machinery. Disassembly of BAK's tertiary structure, such as the truncation of the α1 helix, leads to deregulation of the pro-apoptotic functions and reduction of the protein's stability, thus being implicated in human malignancies. Although many studies have already clarified the vital role of BAK in cellular mechanisms, its pre-mRNA maturation process under cancerous and physiological human cells is neglected. In the present work, we developed and employed a custom multiplexed nanopore sequencing approach that enabled the identification and structural characterization of previously undescribed BAK1 mRNA transcripts (BAK1 v.2-v.11). The described novel mRNAs are derived from multiple types of alternative splicing events, including exon skipping and intron retentions. The implemented multiplexed long-read sequencing approach provided the detailed expression profile of the novel mRNAs in a wide panel of human malignancies and at the same time allowed their relative quantification as compared to the annotated BAK1 v.1. The validation of each novel transcript was carried out with qPCR-based assays. Our results strongly support that most of the novel BAK1 mRNAs harbor open reading frames with conserved BH domains that provide new insights into the correlated mechanisms of apoptosis suppression and cancer. The current study highlights for the first time the hidden aspects of BAK1's transcriptional landscape in both physiological and cancerous human cells and distinguishes the amino acid sequence of the putative BAK isoforms that may possess key apoptosis-related functions not only in diseases, but also under normal cellular conditions.

The effects of the esterified Quercetin with omega3 and omega6 fatty acids on viability, nanomechanical properties, and BAX/BCL-2 gene expression in MCF-7 cells.

Quercetin is one of the major flavonoids and it appears to have cytotoxic effects on various cancer cells through regulating the apoptosis pathway genes such as BAX and BCL2. Combination of Quercetin (Q) with other compounds can increase its effectiveness. In the present study, the effects of the Quercetin and its esterified derivatives on viability, nanomechanical properties of cells, and BAX/BCL-2 gene expression were investigated. Using the MTT and flow cytometry assays, the cytotoxic potential, apoptosis, and necrosis were investigated. The AFM assay was performed to find the nanomechanical properties of cells as the elastic modulus value and cellular adhesion forces. The BAX/BCL2 gene expression was investigated through the Real-Time PCR. The results showed that the esterification of Quercetin with linoleic acid (Q-LA) and α-linolenic acid (Q-ALA) increased the cytotoxic potential of Q. The elastic modulus value and cellular adhesion forces were increased using the esterified derivatives and the highest ratio of BAX/BCL2 gene expression was observed in Q-LA. Esterified Quercetin derivatives have a higher cytotoxic effect than the un-esterified form in a dose-dependent manner. Esterified derivatives caused the nanomechanical changes and pores formation on the cytoplasmic membrane. One of the internal apoptosis pathway regulation mechanisms of these compounds is increasing the BAX/BCL2 gene expression ratio.

[Expression and clinicopathological significance of Bcl - 2 and Bax genes in colorectal cancer patients complicated with schistosomiasis].

OBJECTIVE: To investigate the expression and clinicopathological significance of Bcl-2 and Bax genes in colorectal cancer (CRC) patients complicated with schistosomiasis. METHODS: The CRC patients receiving surgical treatment in the First Affiliated Hospital of Dali University from June 2016 to June 2020 were recruited as the study subjects, and 30 subjects were randomly sampled from the CRC patients complicated with schistosomiasis (CRC-S group) and 30 subjects were randomly sampled from the CRC patients without schistosomiasis (CRC group) using a random number table method. The cancer specimens were sampled from subjects in the CRC-S and CRC groups, and the peri-cancer specimens were sampled from subjects in the CRC group. The Bcl-2 and Bax expression was quantified in cancer and peri-cancer specimens using a real-time fluorescent quantitative PCR (qPCR) assay and immunohistochemistry at transcriptional and translational levels, and the cell apoptosis was detected in cancer specimens using HE staining. RESULTS: A total of 60 subjects were enrolled, including 30 cases in the CRC group and 30 cases in the CRC-S group. There were no significant differences between the two groups in terms of gender distribution (χ2 = 0.271, P > 0.05), mean age (t = -0.596, P > 0.05), tumor growth pattern (χ2 = 0.275, P > 0.05), tumor location (χ2 = 4.008, P > 0.05), tumor invasion depth (χ2 = 0.608, P > 0.05), degree of tumor differentiation (χ2 = 0.364, P > 0.05), or presence of vascular metastasis (χ2 = 1.111, P > 0.05), while significant differences were seen between the two groups in terms of histological type, presence of lymph node metastasis and TMN staging (χ2 = 5.963, 8.297 and 5.711, all P values < 0.05). qPCR assay and immunohistochemistry quantified significantly higher Bcl-2 and Bax expression in cancer specimens from the CRC and CRC-S groups than in the peri-cancer specimens from the CRC group at both translational and transcriptional levels (all P values < 0.05), and higher Bcl-2 and lower Bax expression were seen in the cancer specimens from the CSC-S group than that from the CRC group (all P values < 0.05). In addition, the cell apoptotic rate was significantly greater in the cancer specimens in the CRC group than in the CRC-S group (42.00% vs. 23.35%; χ2 = 41.500, P = 0.000). CONCLUSION: Schistosomiasis may be involved in the development and progression of CRC through affecting Bcl-2 and Bax gene expression in the apoptosis signaling pathway.

The diagnostic significance of 64-slice spiral CT combined with serological CA19-9, Bcl-2, CYFRA21-1 detection in thoracic esophageal carcinoma.

BACKGROUND: To analyze the clinical value of multi-slice spiral computed tomography (MSCT) combined with carbohydrate antigen 19-9 (CA19-9), B-cell leukemia/lymphoma-2 protein (Bcl-2), and cytokeratin 19 fragment antigen 21-1 (CYFRA21-1) detection in the diagnosis of thoracic esophageal cancer. METHODS: The clinical data of 74 patients with thoracic esophageal cancer admitted to the Dazu District People's Hospital in Chonqing, China, from December 2019 to December 2020 were collected (esophageal cancer group), and their computed tomography (CT) signs were analyzed. Another 55 healthy people who underwent physical examination during the same period in the hospital were selected for the healthy group. The serum levels of CA19-9, Bcl-2, and CYFRA21-1 in the different populations were compared, using the receiver operating characteristic (ROC) curve to analyze the value of MSCT combined with CA19-9, Bcl-2, and CYFRA21-1 detection in the diagnosis of thoracic esophageal cancer. RESULTS: The serum levels of CA19-9, Bcl-2, and CYFRA21-1 in patients of the esophageal cancer group were significantly higher than those in the healthy group (P<0.05). The serum levels of CA19-9, Bcl-2, and CYFRA21-1 in patients with poorly differentiated, stage III-IV carcinoma and lymph node metastasis were significantly higher than in those patients with moderately well-differentiated, stage I-II carcinoma and no lymph node metastasis (P<0.05). The CT scans of patients in the esophageal cancer group showed esophageal walls with irregular, needle-shaped, circular, or localized eccentric thickening and narrowed lumens, which were dilated above the cancerous lesions. Some tumors compressed adjacent organs to deform and shift the organs, resulting in the disappearance of surrounding fat layers. Enhanced scans showed mild or moderate enhancement, with large-diameter lesions unable to enhance central, low-density, necrotic areas. The ROC curve showed that the area under the curve (AUC) and the sensitivity and specificity of MSCT combined with CA19-9, Bcl-2, and CYFRA21-1 detection were all higher than for esophageal lesions detected by individual indicators. CONCLUSIONS: CA19-9, Bcl-2, and CYFRA21-1, which are abnormally expressed in patients with esophageal cancer, may be related to the occurrence and development of esophageal cancer. MSCT combined with CA19-9, Bcl-2, and CYFRA21-1 detection appears to enhance the diagnosis of esophageal cancer.

Expression and clinicopathological significance of Bcl - 2 and Bax genes in colorectal cancer patients complicated with schistosomiasis / 中国血吸虫病防治杂志

Objective To investigate the expression and clinicopathological significance of Bcl-2 and Bax genes in colorectal cancer (CRC) patients complicated with schistosomiasis. Methods The CRC patients receiving surgical treatment in the First Affiliated Hospital of Dali University from June 2016 to June 2020 were recruited as the study subjects, and 30 subjects were randomly sampled from the CRC patients complicated with schistosomiasis (CRC-S group) and 30 subjects were randomly sampled from the CRC patients without schistosomiasis (CRC group) using a random number table method. The cancer specimens were sampled from subjects in the CRC-S and CRC groups, and the peri-cancer specimens were sampled from subjects in the CRC group. The Bcl-2 and Bax expression was quantified in cancer and peri-cancer specimens using a real-time fluorescent quantitative PCR (qPCR) assay and immunohistochemistry at transcriptional and translational levels, and the cell apoptosis was detected in cancer specimens using HE staining. Results A total of 60 subjects were enrolled, including 30 cases in the CRC group and 30 cases in the CRC-S group. There were no significant differences between the two groups in terms of gender distribution (χ2 = 0.271, P > 0.05), mean age (t = -0.596, P > 0.05), tumor growth pattern (χ2 = 0.275, P > 0.05), tumor location (χ2 = 4.008, P > 0.05), tumor invasion depth (χ2 = 0.608, P > 0.05), degree of tumor differentiation (χ2 = 0.364, P > 0.05), or presence of vascular metastasis (χ2 = 1.111, P > 0.05), while significant differences were seen between the two groups in terms of histological type, presence of lymph node metastasis and TMN staging (χ2 = 5.963, 8.297 and 5.711, all P values < 0.05). qPCR assay and immunohistochemistry quantified significantly higher Bcl-2 and Bax expression in cancer specimens from the CRC and CRC-S groups than in the peri-cancer specimens from the CRC group at both translational and transcriptional levels (all P values < 0.05), and higher Bcl-2 and lower Bax expression were seen in the cancer specimens from the CSC-S group than that from the CRC group (all P values < 0.05). In addition, the cell apoptotic rate was significantly greater in the cancer specimens in the CRC group than in the CRC-S group (42.00% vs. 23.35%; χ2 = 41.500, P = 0.000). Conclusion Schistosomiasis may be involved in the development and progression of CRC through affecting Bcl-2 and Bax gene expression in the apoptosis signaling pathway.

[The NOXA promoter could function as an active enhancer to regulate the expression of BCL2 in the apoptosis response].

The transcription of eukaryotic genes is regulated by both proximal promoters and distal enhancers. Some promoters also have enhancer activity. NOXA and BCL2 are pro-apoptotic and anti-apoptotic members of the BCL2 family of protein, respectively. Our previous study has found that the NOXA gene promoter and the BCL2 gene promoter interact at the level of three-dimensional chromatin structure. Moreover, the NOXA gene promoter region displays histone modifications characteristic of both promoters and enhancers. This study aimed to explore whether and when the NOXA promoter could act as an active enhancer to regulate BCL2 expression. Based on the apoptosis model of MCF-7 cells induced by camptothecin, we used chromosome conformation capture (3C), quantitative real-time PCR (qRT-PCR) and the luciferase reporter gene technology to demonstrate that the NOXA promoter could function as an active enhancer and physically interact with the BCL2 promoter through chromatin looping. The regulatory properties of the NOXA promoter were closely related to the strength of the apoptosis stimulation. Under weak apoptotic stimulation (1 µmol/L camptothecin treatment), the NOXA promoter mainly functioned as an enhancer; with the enhancement of apoptotic stimulation (10 µmol/L camptothecin treatment), the NOXA promoter activity increased and mainly regulated the expression of the gene itself to promote apoptosis. Chromatin immunoprecipitation (ChIP) confirmed that the dynamic changes of the promoter activity and enhancer activity in the NOXA promoter region are consistent with its histone modification marks. This study provides new clues for further exploring the mechanism underlying cooperative response of BCL2 family member to apoptosis stimuli.

The Healing Effect of Aloe Vera Gel on Acetic Acid-Induced Ulcerative Colitis in Rat.

BACKGROUND The use of herbal and synthetic compounds can be effective in improving the areas and repair of tissues that have been affected during the processes like what happens in ulcerative colitis (UC) as a common inflammatory disorder. According to the beneficial effects of aloe vera, in this study, we aimed to assess the therapeutic effects of oral aloe vera gel on acetic acid-induced colitis in rats by histopathological and molecular analysis of Bax, and BCL-2 genes expression (using RT-PCR technique) in colon tissue samples. METHODS This experimental study comprised 32 adult male Sprague Dawley rats weighting 220 ± 20 g that were randomly divided into four groups as follows. The control group (healthy rats), colitis group in which UC was induced by transrectal administration of 3% acetic acid with no treatment, oral form of sulfasalazine group in which UC was induced by transrectal administration of 3% acetic acid, then was treated by oral administration of sulfasalazine 500 mg/kg body weight, and the fourth group which received oral form of aloe vera gel (200 mg / kg) for 21 days, respectively after induction of UC. Then, the therapeutic effects of treatment groups were compared with the control group and the colitis group with no treatment, by the assessment of histopathological and molecular changes in the colon tissues of rats on the 7th, 14th, and 21st days. Finally, the collected data were analyzed using statistical tests. RESULTS Histologically, aloe vera gel treatment could reduce and heal colon tissue damages in induced colitis. Also, this gel reduced apoptosis in rat's colon with acetic acid-induced colitis, which showed in significantly decreased in Bax mRNA expression and significantly increased BCL-2 mRNA expression compared with the colitis group with no treatment. CONCLUSION Aloe vera gel has a significant effect on the treatment of UC in rat because of the beneficial effect that was found from aloe vera such as decreasing the severity of colitis as evidenced by histopathological findings, and with respect to apoptosis and gene expression that were related to wound healing process, and suppression of the elevation of Bax mRNA with the upregulation of Bcl-2, which can be considered effective in the treatment of UC.

Overexpression of OCT-1 gene is a biomarker of adverse prognosis for diffuse large B-cell lymphoma (DLBCL): data from a retrospective cohort of 77 Brazilian patients.

BACKGROUND: OCT-1 gene is a member of the POU-homeodomain family of transcriptional regulators of B-lymphocyte differentiation by controlling expression of B-cell specific genes. BCL-2 gene is a potent inhibitor of apoptosis and it is essential during B-cell differentiation into germinal center. These genes may be expressed in diffuse large B-cell lymphoma (DLBCL), but the role of BCL-2 in its prognosis has been contradictory, and OCT-1 has yet to be tested. METHODS: In this study, we aimed to investigate the prognostic impact of OCT-1 and BCL-2 expression in DLBCL treated in the real world with immunochemotherapy in a single center. BCL-2 and OCT-1 genes were available in 78.5% (77/98) DLBCL patients, and the RNA for quantitative real-time PCR was isolated from formalin-fixed paraffin-embedded samples. The values obtained for gene expression were transformed in categorical variable according to their median. RESULTS: Cohort median age was 54.5 years (15-84), 49 (50%) were male, 38/77 (49.4%) and 40/77 (51.9%) presented OCT-1 and BCL-2 expression ≥ median, respectively. The overall response rate (ORR) in all patients was 68.4% (67/98), 65,3% (64/98) of patients acquired complete response, and 3.1% (3/98) partial response, while 6.1% (6/98) were primary refractory. The median follow-up was 3.77 years (95% CI: 3.2-4.1), with 5.43 (95% CI: 2.2-NR) of overall survival (OS) and 5.15 years (95% CI: 2.9-NA) of progression free survival (PFS). OCT-1 ≥ median was associated with shorter OS at univariate analysis (p = 0.013; [HR] 2.450, 95% CI: 1.21-4.96) and PFS (p = 0.019; [HR] 2.270, 95%CI: 1.14-4.51) and BCL-2 gene overexpression presented worse PFS (p = 0.043, [HR] 2.008, 95% CI: 1.02-3.95). At multivariate analysis, OCT-1 overexpression was associated with poor PFS (p = 0.035, [HR] 2.22, 95% CI: 1.06-4.67). CONCLUSION: In this study, we showed that overexpression of OCT1 gene was an independent prognostic factor of adverse outcomes in DLBCL.

Dauricine inhibiting the cell proliferation and inducing the cell apoptosis of human pancreatic cancer cells line SW1900 / 解剖学报

Objective To discuss the proliferation inhibition and apoptosis induction of human pancreatic cancer cell line SW1900 by dauricine and its possible mechanism. Methods The MTT colorimetric method was used to detect the inhibitory effects of cell viability. The apoptosis rate was tested by the Annexin Ⅴ-FITC / PI fluorescent staining of flow cytometric method . The expressions of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and B-cell lymphoma-2 (Bcl-2) were detected by Real-time PCR and Western blotting assay. Results MTT assay showed that dauricine significantly inhibited the proliferation of SW1900 cells and the inhibitory effect was enhanced with the increasing of dauricine concentration, F = 783. 7, P < 0. 001. The apoptosis of 3 groups cells were (4. 34 ± 1. 30) % (0 mg / L dauricine), (14. 94±1. 94) % (6 mg / L dauricine) and (22. 68±3. 61) % (12 mg / L dauricine) . The mean difference was statistically significant among the three groups (F = 58. 52, P < 0. 001) . Dauricine could significantly induce apoptosis human pancreatic cancer cells with dose-dependent manner. Real-time PCR showed that the gene expressions of PI3K, Akt and Bcl-2 were lower obviously (PI3K mRNA, F = 101, P = 0. 01; Akt mRNA, F = 1666, P < 0. 01; Bcl-2 mRNA, F = 753, P<0. 001) with dose-dependent manner. Western blotting assay also showed that the protein expression of PI3K, Akt and Bcl-2 was down-regulated with dose-dependent manner. Conclusion Dauricine has proliferation inhibition and apoptosis inducement effect on human pancreatic cancer cells line SW1900. This function may be concerned with the regulation of PI3K / Akt signal pathway and lower Bcl-2 expression.

Expression of Bcl-2 and Bax genes in peripheral blood lymphocytes of depressed and nondepressed individuals.

BACKGROUND: Involvement of the immune system is one of the issues raised in the pathophysiology of depression. BCL2 and BAX genes are related to immune system regulation. We investigated the BCL2 and BAX expression as a probable mechanism of immune system involvement in depression. MATERIALS AND METHODS: This case-control study was conducted on 28 patients with major depression (case) and 28 nondepressed individuals (control) within the age range of 18-55 years in the Isfahan University of Medical Sciences. Clinical interviews, based on the Diagnostic and Statistical Manual of Mental Disorders, were conducted to detect depression, and Beck's Depression Inventory was used to measure the severity of depression in the individuals. In addition, a real-time polymerase chain reaction was employed to compare the level of Bax and Bcl-2 gene expression in peripheral blood lymphocytes. The multivariate covariance analysis was used to explore the correlation between BCL2 and BAX gene expression and to control the effect of duration and severity of depression. RESULTS: The results showed that none of the variables including group membership, the duration of depression, and the severity of depression were not significantly correlated with the expression of BCL2 and BAX genes. Furthermore, there was no statistically significant relationship between the Bax and Bcl-2 genes expression in case and control groups (P > 0.05). CONCLUSION: Depression may have no impact on Bax and Bcl-2 gene expression in patients with major depression. Studies with larger sample size are recommended.

Evaluation of BAX and BCL-2 Gene Expression and Apoptosis Induction in Acute Lymphoblastic Leukemia Cell Line CCRFCEM after High- Dose Prednisolone Treatment

Objective: Glucocorticoids are one of the most important drugs in the treatment of acute lymphoblastic leukemia for children. It is very important to response to glucocorticoid in the prognosis of these patients. Therefore, resistance to treatment is a major problem in lymphoid leukemia cases. In, this study, CCRF-CEM cell line was selected as a chemotherapy-resistant model. The aim of this study was to evaluate the effect of high dose prednisolone on induction of apoptosis and changes in BAX and BCL-2 gene expression at different times. Methods: CCRF-CEM cell lines were grown in standard conditions. Based on previous studies, a dose of 700 µM as subtoxic dose was selected. Changes in gene expression of BAX and BCL-2 were evaluated by using real time PCR techniques. Also stained with annexin V and the induction of apoptosis was assessed by FACS machine. Results: In this study it was found that prednisolone in high doses at different times significantly increased the gene expression of BAX and on the other hand the amount of BCL-2 expression was reduced. Cells that treated for 48 hours had more significant changes in gene expression. Based on flowcytometry data, prednisolone can induce apoptosis in a time dependent manner on this cancerous resistant cell line. Conclusions: Apoptosis induced by high-dose prednisolone in the CCRF-CEM cells, which is almost resistant, and possibly mediated by reducing the expression of BCL-2 and BAX up-regulation.

Codelivery of temozolomide and siRNA with polymeric nanocarrier for effective glioma treatment.

BACKGROUND: The treatment of glioma remains a challenge because conventional chemotherapy is often ineffective by drug resistance. Combinative therapy using chemotherapeutic agents and siRNA has demonstrated potential to improve anticancer outcome through a synergistic effect in various cancers. The current study aims to achieve better glioma treatment through a combinative therapy based on a folate-targeted nanocarrier carrying both temozolomide (TMZ) and anti-BCL-2 siRNA. METHODS: A polymeric micelle (TMZ-FaPEC@siRNA) incorporating TMZ and anti-BCL-2 siRNA was prepared based on folate-conjugated triblock copolymer (Fa-PEG-PEI-PCL, Fa-PEC) of poly(ε-caprolactone) (PCL), poly(ethylenimine) (PEI) and poly(ethylene glycol) (PEG). The physicochemical properties and drug release profile of TMZ-FaPEC@siRNA were tested. The Fa-targeted drug delivery and joint effect of siRNA and TMZ to induce glioma apoptosis and tumor growth inhibition were evaluated both in vitro and in vivo. RESULTS: In vitro cell study demonstrated that the nanocarrier effectively facilitates codelivery of siRNA and TMZ into C6 cells, resulting in a strong apoptotic response of cancer cells by silencing the antiapoptosis BCL-2 gene and activating the proapoptotic Bax gene simultaneously. In vivo study in rat bearing orthotropic glioma showed that tumor growth was inhibited and meanwhile animal survival was prolonged remarkably through intracranial injection of TMZ-FaPEC@siRNA. CONCLUSION: Our results evidence the strong efficacy of the folate-targeted nanomedicine carrying TMZ and BCL-2 siRNA in treating glioma.

Cytotoxic and apoptotic potential of Myristica fragrans Houtt. (mace) extract on human oral epidermal carcinoma KB cell lines

Several studies have revealed that certain naturally occurring medicinal plants inhibit the growth of various cancers. The present study was conducted to evaluate cytotoxicity and apoptotic induction potential of Myristica fragrans Houtt mace extract. The cytotoxic activity of the Myristica fragrans Houtt mace acetone extract was assayed by MTT assay on human oral epidermal carcinoma KB cell lines. KB cells were incubated with different concentration of mace extract ranging from 25 to 125 µg/mL for 24hrs. The apoptotic induction potential was also studied by the analysis of Bcl-2 protein and gene expression in mace extract incubated KB cell lines using western blotting technique and real-time polymerase chain reaction. The mace extract exhibited cytotoxicity and anticancer effect against KB cell lines and it also suppressed the growth of cancer cells, therefore growth inhibitory effect was noted in extract treated cell lines. The apoptotic potential of mace extract was accompanied by reduced gene expression of Bcl-2 compared to the untreated KB cells. The mace extract shows the cytotoxic activity and induced the apoptosis through the modulation of its target genes Bcl-2 in the KB cell lines, suggesting the potential of mace as a candidate for oral cancer chemoprevention. This can be further investigated in vivo for its anticancer potential.