• Journal of Microbiology and Biotechnology
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• v.18 no.9
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• pp.1590-1598
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• 2008

Cell proliferation and differentiation are critical processes in a developing fetal rat brain, during which programmed cell death (PCD) also plays an important role. One of the decisive factors for PCD is Bcl-2 family proteins, where Bax induces cell death, whereas Bcl-2 acts as an inhibitor of PCD. As maternal drinking is known to cause fetal alcohol syndrome (FAS) or malformation of the fetal brain during pregnancy, the objective of the present study was to investigate whether maternal ethanol exposure alters the PCD-related Bax and Bcl-2 protein expression during fetal brain development. Pregnant female rats were orally treated with 10% ethanol and the subsequent expressions of the Bax and Bcl-2 proteins examined in the fetal brain, including the forebrain, midbrain, and hindbrain, from gestational day (GD) 15.5 to GD 19.5, using Western blots, in situ hybridization, and immunohistochemistry. With regard to the ratio of Bcl-2 to Bax proteins (Bcl-2/Bax), the Bax protein was dominant in the forebrain and midbrain of the control GD 15.5 fetuses, except for the hindbrain, when compared with the respective ethanol-treated groups. Moreover, Bcl-2 became dominant in the midbrain of the control GD 17.5 fetuses when compared with the ethanol-treated group, representing an alternation of the natural PCD process by ethanol. Furthermore, a differential expression of the Bcl-2 and Bax proteins was found in the differentiating and migrating zones of the cortex, hippocampus, thalamus, and cerebellum. Thus, when taken together, the present results suggest that ethanol affects PCD in the cell differentiation and migration zones of the prenatal rat brain by modulating Bax and Bcl-2 expression in an age- and area-dependent manner. Therefore, this is the first evidence that ethanol may alter FAS-associated embryonic brain development through the alteration of Bax and Bc1-2 expression.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.3
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• pp.167-178
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• 2002

Objective : The present study was performed to investigate whether apoptosis occur in human embryos by annexin staining and detect the expression of Fas, Fas-ligand (FasL), Bax, and Bcl-2 in human fragmented embryos derived from IVF-ET by immunofluorescence and Western blot analysis. Materials and Methods: Using annexin staining, immunofluorescence and Western blot analysis on normal and fragmented embryos, we were able to detect apoptotsis and apoptotic gene products in fragmented embryos. Result: Phosphatidylserine (PS) translocation, the marker for apoptosis, were detected frequently in fragmented embryos. Bcl-2 and Bax protein were detected in both fragmented and non-fragmented embryos. When fragmented embryos compared to normal embryos, immunofluorescent intensity of Bcl-2 tended to be lower in fragmented embryos. Bax gene expression increased in the fragmented embryos compared to the normal embryos. This result supports a model in which the molar ratio of Bcl-2 to Bax determines whether apoptosis induced or inhibited in human embryo. Fas was highly expressed in human preimplantation embryos but not FasL. It suggests that embryo may undergo apoptosis by binding with FasL produced by follicular or immune cells. Conclusion: The over expression of Bax and Fas will trigger apoptosis to lead embryo fragmentation and change embryo to be nonviable.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.6
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• pp.671-676
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• 2006

We investigated the effect of gingerol (Zingiber officinale Roscoe, Zingiberaceae) on Bcl-2 and Bax expression in MDA-MB-231 human breast cell lines. The oleoresin from rhizomes of ginger contains [6]-gingerol (1-[4'-hydroxy-3'-methoxyphenyl]-5-hydroxy-3-decanone). We previously reported that [6]-gingerol inhibits cell proliferation in MDA-MB-231 human breast cancer cell lines. In this study, we examined protein and mRNA expression associated with cell apoptosis in MDA-MB-231 human breast cancer cell lines. We cultured MDA-MB-231 cells in presence of various concentrations 0, 2.5, 5 and $10\;{\mu}M$ of [6]-gingerol. Bcl-2 protein and its mRNA levels were decreased dose-dependently in cells treated with [6]-gingerol, but Bax protein and its mRNA levels were unchanged by [6]-gingerol treatment. Bcl-2/Bax ratio was decreased in a dose dependent manner treated with [6]-gingerol. Caspase-3 activity was significantly increased dose-dependently in cell treated with [6]-gingerol (p<0.05). In conclusion, we have shown that [6]-gingerol induces apoptosis in MDA-MB-231 human breast cancer cell lines.

• Radiation Oncology Journal
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• v.23 no.2
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• pp.98-105
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• 2005

Purpose : The objective of our study was to evaluate the immunohistochemical expression of p53 and bax proteins as prognostic markers in FIGO stage IIb invasive squamous cell carcinoma of the uterine cervix. Materials and Methods : Sixty-five cases of squamous cell carcinoma of the cervix (stage IIb) that were diagnosed from October 1995 to December 2003 were analyzed retrospectively for the bax and p53 expression. These expressions were determined immunohistochemically and they were correlated to the patients' overall survival and disease-free survival. Results : The overall 5-year survival (OS) rate and the disease-free survival (DFS) rate were $65.1\%$ and $62.9\%$, respectively. p53 and bax immunoreactivity was seen in $26.2\%$ and $52.3\%$ of cases, respectively, with variable levels of expression. On the univariate analysis, only p53 positivity correlated with poor survival in DFS (log-rank test p=0.027), but this significance was not maintained on multivariated analysis by Cox's regression. The nine cases with the immunophenotype ps3+/bax- had the poorest survival. Conclusion : Neither p53 nor bax expression are Independent predictors of the prognosis for stage IIb cervical squamous cancers. Evaluation of p53 and bax co-expression may affect the clinical outcome and further investigation is needed.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.5
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• pp.389-396
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• 2001

Neuroprotective properties of lithium were investigated by using in vivo NMDA excitotoxicity model. The appearance of TUNEL positive cells was prominent within 24 h of NMDA (70 mg/kg, i.p.) injection in the regions of the cortex, hippocampal formation, and thalamus of mouse cerebrum. NMDA treatment resulted in the extensive enhancement of Bax immunoreactivity in the cortical and hippocampal regions. NMDA also increased the immunoreactivity of caspase 8 in the similar regions of the mouse cerebrum. However, the increased immunoreactivity of Bax and caspase 8 were dramatically attenuated by chronic lithium pretreatment (lithium chloride, 300 mg/kg/d, i.p. for $7{\sim}10$ days). At the same time, lithium ion blocked the appearance of TUNEL positive cells, and the morphological assessment indicated an effective neuroprotection by lithium against NMDA excitotoxicity. Although the exact action mechanism of lithium is not straightforward at this time, we propose that the inhibition of Bax and caspase cascade is involved in the neuroprotective action of lithium.

• The Journal of the Korean bone and joint tumor society
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• v.11 no.2
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• pp.118-125
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• 2005

Purpose: The expression of apoptosis-related genes, such as survivin, bcl-2, and bax has been examined in the human osteosarcoma and then evaluated the correlation with clinical data of patients. Materials and Methods: Fifty human osteosarcoma specimens were established from incisional biopsy and examination of survivin, bcl-2, and bax by immunohistochemical study was performed. We investigated the correlation of survivin, bcl-2, bax and their two or three combined expressions with clinical data including the response of chemotherapy, local recurrence, distance metastasis, and oncologic outcome. Results: Survivin was showed in 26 cases (52%), bcl-2 in 23 cases (46%), and bax in 21 cases (42%) osteosarcoma. And coexpression of survivin and bcl-2 was showed in 19 cases (38%), survivin and bax in 13 cases (26%), bcl-2 and bax in 8 cases (16%), and all three expression was showed in 8 cases (16%). There was no correlation between their apoptosis related gene and histologic difference, the presence of local recurrence and distant metastasis. Whereas neoadjuvant chemotherapy response correlated with bcl-2 expression (P=0.04), and survivin and bcl-2 coexpression (P=0.044) with poor chemoresponse. The rate of died of disease was correlated with bcl-2 (P=0.001), survivin and bcl-2 coexpression (P=0.027) with bad outcome. Survival curves of bcl-2 (P=0.0075), survivin and bcl-2 (P=0.0012) was showed negative correlation in the Kaplan-Meier method. Conclusion: The apoptosis related gene expression was relatively high in osteosarcoma, bcl-2 expression was correlated with poor chemotherapy response and poor survival rate, but survivin was correlated with this oncologic outcome only in the bcl-2 coexpression. The examination of immunohistochemical stain of apoptosis related gene in osteosarcoma could be helpful in the judgment of osteosarcoma prognosis.

• Molecules and Cells
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• v.24 no.3
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• pp.378-387
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• 2007

The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

• Journal of Korean Medicine Rehabilitation
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• v.20 no.1
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• pp.1-12
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• 2010

Objectives : Cool Pharmacopuncture for static blood is a famous pharmacopuncture treatment that treated disease caused by static blood. Hyolhae(Sp10) is also a famous point of acupuncture that treated that. This study was designed to evaluate the effects of Cool Pharmacopuncture for static blood into Hyolhae(Sp10) on BAX and BCL-2 expression in the experimental traumatic brain injury(TBI) rats. Methods : Male rats were divided into 3 groups. I was no treatment after TBI. II was treatment with needle-prick acupuncture after TBI. III was treatment with Cool Pharmacopuncture for static blood into Hyolhae(Sp10) after TBI. The author carried out neurological motor behavioral test, histological assessment test. Neurological motor behavior tests consist of rotarod test, beam-walking test and postural reflex test. In the histological assessment test, BAX and BCL-2 expression, hematoxylin & eosin staining were experimented. Results : In neurological motor behavior tests, motor and cognitive function recovery was significantly increased in the II, III as compared with I (p<0.05). Especially III was significantly increased as compared with II (p<0.05). BAX expression was significantly decresed in order of the III, II, I after 7 and 14 days later. BCL-2 expression was investigated in the III, II as compared with I. Especially Most incresed expression was experimented in the III. Conclusions : According to the above results, Cool Pharmacopuncture for static blood can inhibit apoptosis of cells after TBI in rats by contol of BAX and BCL expression.

• Korean Journal of Head & Neck Oncology
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• v.16 no.2
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• pp.161-166
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• 2000

Objective: The nm23 gene has been identified as a potential metastasis suppressor gene in various human neoplasms. Both Bcl-2, which promotes cell survival, and Bax, which promotes cell death, have been considered as major factors in controlling the apoptotic pathway. This study was carried out to determine whether these markers are useful in distinguishing potential intrinsic differences in tumor virulence of papillary thyroid cancers. Material and Method: The expressions of nm23, Bcl-2 and Bax have been evaluated using immunohistochemical techniques in 100 pure papillary thyroid cancers and 20 metastatic lymphnodes. The intensity of immnunoreactivity was graded on arbitrary four point scale(grade 0 or 1 : negative reactivity, grade 2 or 3 positive reactivity). The immunoreactivities were analyzed in relation to TNM atage, AMES score, local recurrence and distant metastasis, and that of metastatic LNs was compared with the tumors. Results: The expression of Bcl-2 and bax did not show any statistical differences by TNM stage, AMES score, recurrence, distant metastasis and also between the tumor and metastatic LN. However, the nm23 showed higher expression of Ki67 in distant metastasis than in control group and in metastatic LNs than in the tumors(p<0.05). Conclusion: Although the expression of Bcl-2 and Bax protein showed no correlation with clinical parameters representing tumor virulence, the nm23 expression could be an useful prognostic factor, especially in predicting nodal or distant metastasis in papillary thyroid cancer.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.155-160
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• 2020

Objective: The genes Bcl-2 and Bax play important roles in apoptosis. Many studies have shown that formalin has a strong deleterious effect on male fertility and can induce apoptosis. L-carnitine has been reported to potentially reverse the negative effects of formalin, leading to improved spermatogenesis. In this study, we examined the levels of expression of Bcl-2 and Bax in mice treated with formalin and L-carnitine. Methods: Thirty adult BALB/c mice were categorized into three groups. The mice in the control group (n = 10) were not injected with any substance. The mice in the second group (n = 10) received 10 mg/kg of formalin daily via an intraperitoneal injection, while those in the final group (n = 10) were intraperitoneally injected daily with a dose of 10 mg/kg of formalin and 100 mg/kg of L-carnitine. All mice were kept in isolated cages for 31 days. Results: The expression of Bax was significantly higher in the formalin-treated mice than in the mice of the control group, while the expression of Bcl-2 was significantly lower in the formalin-treated mice than in the control mice. Additionally, relative to control mice, Bcl-2 expression increased and Bax expression decreased in the mice administered both formalin and L-carnitine. Conclusion: In this study, L-carnitine was shown to augment Bcl-2 expression and to reduce Bax expression, indicating that this compound may inhibit apoptosis. Due to its positive effects, L-carnitine can be used as a prophylactic treatment for people who routinely come into direct contact with formalin as an occupational hazard.