• Title/Summary/Keyword: p-53

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Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice (아데노바이러스 유전자치료벡터의 생식독성 연구)

  • Rhee, Gyu-Seek;Kwack, Seung-Jun;Kim, Soon-Sun;Lee, Rhee-Da;Seok, Ji-Hyun;Chae, Soo-Young;Chung, Soo-Youn;Kim, Seung-Hee;Lee, Seung-Hoon;Park, Kui-Lea
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.151-158
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    • 2007
  • The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

The Immunohistochemical Analysis for the Expression of Survivin, an Inhibitor of Apoptosis Protein, in Non-small Cell Lung Cancer (비소세포폐암에서 아포프토시스 억제 단백질 Survivin 발현에 관한 면역조직학적 분석)

  • Ko, Mi-Hye;Myoung, Na-Hye;Lee, Jae-Whan;Cho, Eun-Mi;Park, Jae-Seuk;Kim, Keun-Youl;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.909-921
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    • 2000
  • Background : Defects in apoptotic signaling pathways play important role in tumor initiation, progression, metastasis and resistance to treatment. Several proteins which may promote tumorigenesis by inhibiting apoptosis were identified. The survivin protein is the member of inhibitor of apoptosis protein(IAPs) family which inhibits apoptosis. Unlike other IAPs, it is expressed in during the fetal period but not in adult differentiated tissues. Many reports have stated that survivin is selectively expressed in many cancer cell lines and cancer tissues. We performed immunohistochemical analysis for survivin expression in non-mall cell lung cancer to get evaluate its clinical implication. Methods : Twenty nine surgically resected lung cancers were examined. Immunohistochemical staining were performed by immuno-peroxidase technique using avidin-biotinylated horseradish pemxidase complex in the formalin-fixed, paraffin-embedded tissue $4{\mu}m$ section. Anti-survivin polyclonal antibody was used for primary antibody and anti-p53 monoclonal antibody was also used to analyze the correlation between survivin and p53 expression. The survivin expression scores were determined by as the sum of the stained area and intensity. Results : Immunohistochemical analysis showed cancer specific expression of survivin in 20 of 29 cases (69.0%). Western blot analysis also showed the selective survivin expression in tumor tissue. There was no correlation between survivin expression and clinicopathological parameters and prognosis. We analyzed the ∞π'elation between survivin expression and p53 expression, but found none. Conclusion: We confirmed the tumor specific expression of survival in non-small cell lung canær. But this expression was not correlated with clinical parameters as well as histology, tumor stage, recurrence, and survival rate. Also it was not statistically correlated with the expression of p53.

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Silence of LncRNA GAS5 Protects Cardiomyocytes H9c2 against Hypoxic Injury via Sponging miR-142-5p

  • Du, Jian;Yang, Si-Tong;Liu, Jia;Zhang, Ke-Xin;Leng, Ji-Yan
    • Molecules and Cells
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    • v.42 no.5
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    • pp.397-405
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    • 2019
  • The regulatory role of long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) in both cancerous and noncancerous cells have been widely reported. This study aimed to evaluate the role of lncRNA GAS5 in heart failure caused by myocardial infarction. We reported that silence of lncRNA GAS5 attenuated hypoxia-triggered cell death, as cell viability was increased and apoptosis rate was decreased. This phenomenon was coupled with the down-regulated expression of p53, Bax and cleaved caspase-3, as well as the up-regulated expression of CyclinD1, CDK4 and Bcl-2. At the meantime, the expression of four heart failure-related miR-NAs was altered when lncRNA GAS5 was silenced (miR-21 and miR-142-5p were up-regulated; miR-30b and miR-93 were down-regulated). RNA immunoprecipitation assay results showed that lncRNA GAS5 worked as a molecular sponge for miR-142-5p. More interestingly, the protective actions of lncRNA GAS5 silence on hypoxia-stimulated cells were attenuated by miR-142-5p suppression. Besides, TP53INP1 was a target gene for miR-142-5p. Silence of lncRNA GAS5 promoted the activation of PI3K/AKT and MEK/ERK signaling pathways in a miR-142-5p-dependent manner. Collectively, this study demonstrated that silence of lncRNA GAS5 protected H9c2 cells against hypoxia-induced injury possibly via sponging miR-142-5p, functionally releasing TP53INP1 mRNA transcripts that are normally targeted by miR-142-5p.

Involvement of p53-Mediated Mitochondrial Stress in the Apoptosis Induced by Flavonoids Purified from Rhus verniciflua Stokes in Human Osteosarcoma Cells

  • Chung, Song-Woo;Lee, Seung-Ah;Park, Jong-Sun;Ryu, Kwon-Woo;Jang, Mun-Ju;Park, Song-Soo;Lee, Choon-Bong;Kim, Jong-Ghee;Jeon, Young-Mi;Lee, Jeong-Chae
    • Natural Product Sciences
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    • v.13 no.1
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    • pp.1-5
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    • 2007
  • Dietary flavonoids have antioxidant and antitumor promoting effects. Rhus verniciflua Stokes (RVS) is a flavonoid-rich herbal medicine and has long been used as a food additive and an antitumor agent in Korea. Previous study demonstrated that a purified flavonoid fraction prepared from RVS, herein named RCMF (the RVS chloroform-methanol fraction), exhibited growth inhibition and induced apoptosis in human osteosarcoma(HOS) cells. This study evaluated if p53-mediated pathway is associated with the RCMF-induced apoptosis in HOS cells. RCMF was shown to be capable of inducing apoptosis of the cells, as expected, and transparently increased p53 expression in the cells. However, the RCMF-induced cytotoxicity was suppressed by transfecting the cells with antisense p53 oligonucleotide, which also inhibited the decrease of Bcl-2 and the increase of Bax in mitochondria, and the release of cytochrome c into cytosol. This finding suggests that p53-mediated mitochondrial stress is required for RCMF-induced apoptosis in HOS cells.

p53 Codon 72 Polymorphism Interactions with Dietary and Tobacco Related Habits and Risk of Stomach Cancer in Mizoram, India

  • Malakar, Mridul;Devi, K. Rekha;Phukan, Rup Kumar;Kaur, Tanvir;Deka, Manab;Puia, Lalhriat;Sailo, Lalrinliana;Lalhmangaihi, T.;Barua, Debajit;Rajguru, Sanjib Kumar;Mahanta, Jagadish;Narain, Kanwar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.717-723
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    • 2014
  • Background: This study was carried out to investigate the interaction of p53 codon 72 polymorphism, dietary and tobacco habits with reference to risk of stomach cancer in Mizoram, India. A total of 105 histologically confirmed stomach cancer cases and 210 age, sex and ethnicity matched healthy population controls were included in this study. Materials and Methods: The p53 codon 72 polymorphism was detected by PCR-RFLP and sequencing. H. pylori infection status was determined by ELISA. Information on various dietary and tobacco related habits was recorded with a standard questionnaire. Results: This study revealed that overall, the Pro/Pro genotype was significantly associated with a higher risk of stomach cancer (OR, 2.54; 95%CI, 1.01-6.40) as compared to the Arg/Arg genotype. In gender stratified analysis, the Pro/Pro genotype showed higher risk (OR, 7.50; 95%CI, 1.20-47.0) than the Arg/Arg genotype among females. Similarly, the Pro/Pro genotype demonstrated higher risk of stomach cancer (OR, 6.30; 95%CI, 1.41-28.2) among older people (>60 years). However, no such associations were observed in males and in individuals <60 years of age. Smoke dried fish and preserved meat (smoke dried/sun dried) consumers were at increased risk of stomach cancer (OR, 4.85; 95%CI, 1.91-12.3 and OR, 4.22; 95%CI, 1.46-12.2 respectively) as compared to non-consumers. Significant gene-environment interactions exist in terms of p53 codon 72 polymorphism and stomach cancer in Mizoram. Tobacco smokers with Pro/Pro and Arg/Pro genotypes were at higher risk of stomach cancer (OR, 16.2; 95%CI, 1.72-153.4 and OR, 9.45; 95%CI, 1.09-81.7 respectively) than the non-smokers Arg/Arg genotype carriers. The combination of tuibur user and Arg/Pro genotype also demonstrated an elevated risk association (OR, 4.76; 95%CI, 1.40-16.21). Conclusions: In conclusion, this study revealed that p53 codon 72 polymorphism and dietary and tobacco habit interactions influence stomach cancer development in Mizoram, India.

Sulforaphane-Induced Apoptosis was Regulated by p53 and Caspase-3 Dependent Pathway in Human Chondrosarcoma, HTB-94 (Sulforaphane에 의한 p53 및 caspase-3 의존 신호전달계를 통한 인간 연골암 세포주 HTB-94에서의 세포사멸 기전 연구)

  • Lee, Won-Kil;Kim, Song-Ja
    • Journal of Life Science
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    • v.21 no.6
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    • pp.851-857
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    • 2011
  • Sulforaphane (SFN) is an isothiocyanate, isolated from glucoraphanin in broccoli and other cruciferaous vegetables. Recent studies have revealed that SFN induces anti-proliferation and apoptosis by cell cycle arrest in various cancer cells. In this study, we investigated the effect of SFN induced apoptosis in chondrosarcoma HTB-94 cells. SFN caused suppression of proliferation and apoptosis in a dose-dependent manner as determined by cell phenotype, MTT assay and FACS analysis in HTB-94 cells. Treatment of SFN led to caspase-3 activation and p53 accumulation as determined by Western blot analysis. Also, SFN significantly induced DNA fragmentation and nuclear degradation though activation of caspase-3, as detected by DNA electrophoresis and immunostaining, respectively. Our results indicate that SFN-induced apoptosis was regulated by p53 and caspase-3 dependent pathways. Furthermore, SFN may act as a potent anti-proliferation agent, and as a promising candidate for molecular-targeting chemotherapy against human chondrosarcoma cells.

Spontaneous Apoptosis and Metastasis in Squamous Cell Carcinoma of the Lung (폐 편평세포암에서 자발성 아포토시스와 원격전이)

  • Oh Yoon-Kyeong;Kee Keun-Hong
    • Radiation Oncology Journal
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    • v.17 no.3
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    • pp.203-208
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    • 1999
  • Purpose : To evaluate whether spontaneous apoptosis has prognostic value among patients with squamous cell carcinoma of lung. Materials and Methods : Material from 19 patients who received thoracic irradiation between 1990 and 1994 was analyzed. Their stages were II (1), IIIa (8), IIIb (5), and IV (5). Patients were observed from 5 to 67 months (median : 17 months). The spontaneous apoptosis index (AI) and p53 mutation were measured by immunohistochemical stains. Results : AI was found to range from 0 to $1\%$ (median $0.4\%$). Patients with low AI ($AI{\leq}$median) had a much higher distant metastasis rate at diagnosis than patients with high AI. By analysis of prognostic factors for survival, M stage was significant in univariate analysis. AI, chemotherapy, M stage, T stage, and stage were significant in multivariate analysis. The correlation between the AI and p53 mutation was not seen. Conclusion : AI was related with distant metastasis at diagnosis and not with p53 mutation. Also low AI group tended to have shorter survival time than high AI group.

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Factors Affecting the Number of Stapler Cartridges in Complete Video-Assisted Thoracoscopic Surgery Lobectomy for Non-small Cell Lung Cancer

  • Yang, Young Ho;Lee, Seokkee;Lee, Chang Young;Kim, Dae Joon;Lee, Jin Gu;Chung, Kyung-Young
    • Journal of Chest Surgery
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    • v.53 no.2
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    • pp.53-57
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    • 2020
  • Background: Video-assisted thoracoscopic surgery (VATS) lobectomy has become the major surgical option for the treatment of non-small cell lung cancer (NSCLC). Endoscopic instruments such as stapler cartridges are essential for VATS procedures. In this study, we investigated the factors that affect the number of stapler cartridges used in VATS lobectomy. Methods: A retrospective analysis was conducted of patients who underwent complete VATS lobectomy for NSCLC from January 2013 to December 2015. Results: In total, 596 patients underwent complete VATS lobectomy. The average number of stapler cartridges used for VATS lobectomy was 5.3±1.9. The number of stapler cartridges used for VATS lobectomy was higher in men (5.5±1.9 vs. 5.0±18, p=0.006), those aged older than 70 years (5.5±2.1 vs. 5.1±1.7, p=0.038), those who underwent upper or middle lobectomy procedures (5.7±1.9 vs. 4.1±1.2, p<0.001), those with a higher fissure sum average (p<0.001), and those in whom surgery was performed by a surgeon with a preference for staplers (5.6±2.0 vs. 4.9±1.6, p<0.001). Conclusion: The number of stapler cartridges required to perform VATS lobectomy in NSCLC patients appears to be influenced by sex, age, the location of the tumor, the degree of fissure development, and the surgeon's preference.