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The impact of ^(60)Co γ-ray on cycles to hep-2 human laryngeal cancer cell in the condition of hypoxia

The Chinese-German Journal of Clinical Oncology 2010年第7期9卷 373-377页

作者： Baoshan Wang Guibo Li Dongmei Song Jihua Zhang Department of Otolaryngology-Head and Neck Surgery The First Hospital Hebei Medical University Shijiazhuang 050031 China Family Planning Service Station of Haigang area Qjnhuangdao 066000 China

Objective： The aim of the study was to investigate the impact of 60Co y-ray on apoptosis, cell cycles and the expression of protein hypoxia-inducible factor-1α （HIF-1α） to hep-2 cell line in the conditions of normoxia and hypoxia. Methods： hep-2 cell were divided into 2 groups： group A （normoxia） and group B （hypoxia）. All of the ceils were exposed to y-ray with dosage being 0, 1, 3, 5, 10, 20, and 40 Gy. Flow cytometry was used to measure the protein level of HIF-1α and to detect apoptosis and cell cycles. The protein level of HIF-1α was also determined by immunohistochemistry and Western blotting. Results： The protein level of HIF-1α in group B was significantly higher than that in group A. In group A, low doses （1-5 Gy） of y-ray had caused G0/G1 cell cycle arrest and high doses （10-40 Gy） had caused G2/M cell cycle arrest. In group B, without exposure of y-ray （0 Gy） had caused G0/G1 cell cycle arrest, all of the different dosage of y-ray could cause G2/M cell cycle arrest. The curve of apoptosis rate in group A was a parabola, the apoptotic rate was related to the dosage of y-ray in a dosage dependent manner. The peak was at the point of 5 Gy. The apoptosis rate in group A was significantly higher than that in group B. Conclusion： Different doses of y-ray could cause different cell cycles arrest then make different impact on apoptosis to hep-2 ceil. The lower apoptosis rate in condition of hypoxia maybe has a relationship with G2/M cell cycle arrest. Up-regulated HIF-1α protein may be one of the reasons for G2/M cell cycle arrest.

关键词： cell cycle y-ray apoptosis hypoxia-inducible factor-1α (HIF-1α) hep-2 cell

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The impact of ^(60)Co γ-ray on apoptosis to hep-2 human laryngeal cancer cell in the condition of reoxygenation after hypoxia

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The Chinese-German Journal of Clinical Oncology 2010年第2期9卷 93-96页

作者： Baoshan Wang Jihua Zhang Dongmei Song Ying Zhang Department of Otolaryngology-Head and Neck Surgery The First Hospital Hebei Medical University Shijiazhuang 050031 China

Objective： The aim of the study was to establish models of hep-2 laryngeal cancer cell line of different oxygen supplying, trying to investigate the impact of normoxia, hypoxia, reoxygenation after hypoxia on apoptosis and expression of proteins HIF-1α and p53 to hep-2 human laryngeal cancer cell line induced by ^60Co γ-ray. Methods： Human laryngeal cancer hep-2 cells were divided into 3 groups： group A （normoxia）, group B （hypoxia）, and group C （reoxygenation after hypoxia）. All of the cells were exposed to 5 Gy dosage of γ-ray. Flow cytometry （FCM） was used tomeasure the protein levels of HIF-1α and p53 and to detect cell apoptosis. The protein levels of HIF-la and p53 were also determined by immunohistochemistry and Western blotting. The expression of HIF-la mRNA was determined by RT-PCR. Results： The protein levels of HIF-1α and p53 were evidently increased in group B compared to group A. The protein levels of HIF-1α and p53 in group C were lower compared to group B; the rate of apoptosis in group C was higher than that in group B. Conclusion： Hypoxia decreased the effect of apoptosis induced by ^60Co γ-ray in hep-2 human laryngeal cancer cell line. The apoptosis pathway maybe related to some other genes or proteins but not p53 in the conditions of hypoxia and reoxygenation after hypoxia.

关键词： γ-ray hypoxia apoptosis p53 hep-2 cell

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Transfection,overexpression and clinical application of human 60 kDa Ro/SSA autoantigens in hep-2 cells

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Chinese Medical Journal 2003年第3期116卷 31-34页

作者：吕良敬陈顺乐顾越英沈南鲍春德王元薛峰叶萍俞翀曌 Shanghai Rheumatology Institute and Department of Rheumatology Renji Hospital of Shanghai Second Medical University Shanghai 200001 China

Objective To develop an improved substrate for indirect immunofluorescence test (IIF) for detecting anti-Ro60/Sjogren's syndrome A (Ro/SSA) *** 60-kDa Ro/SSA autoantigens (Ro60) cDNAs were obtained from human placental cDNA library using polymerase chain reaction (PCR) and were cloned into the mammalian expression vector-pEGFP-C1. Then, the recombinant plasmids were transfected into hep-2 cells. We confirmed the overexpression, localization and antigenicity of fusion proteins in transfected cells by means of immunoblotting, confocal fluorescence microscopy and IIF. hep-2 and hep-Ro60 were analyzed by IIF using a panel of 10 precipitin-positive anti-Ro human sera *** Stable expression of Ro60-green fluorescent protein (Ro60-GFP) fusion proteins were maintained ten more generations. Ro60-GFP kept the antigenicity of Ro while demonstrating its own characteristic immunofluorescent pattern in hep-Ro60 cells. The transfectants dramatically increased the sensitivity of IIF testing (a mean increase of 6.7-fold in endpoint titer). Eight overten (8/10) positive anti-Ro sera showed characteristic immunofluorescent patterns for hep-Ro60, including two sera that were anti-nuclear antibodies (ANA) negative for untransfected hep-2. IIF-ANA in all healthy sera was negative for hep-Ro60. Conclusions As a new substrate for IIF, the Ro60 transfectants can be used to detect anti-Ro antibodies. In addition, transfected hep-2 cells keep the immunofluorescent properties of hep-2 cells in IIF-ANA tests and can be employed as a substrate for routine IIF-ANA detection.

关键词： Ro60/Sjogren's syndrome A transfection hep-2 cell

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Inhibitory Effects of Celecoxib and Sc-58125 on Proliferation of Human Carcinoma of Larynx hep-2 In Vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2005年第2期25卷 202-205页

作者：丁娟常青龚树生 Department of Otolaryngology Union Hospital Tongji Medical College Huazhong Universityof Science and Technology Wuhan 430030 China

Summary: The inhibitory effects of two kinds of selective cyclooxygenase-2 inhibitors on the proliferation of human carcinoma of larynx hep-2 in vitro and their corresponding mechanisms were investigated. hep-2 cells were cultured with two kinds of selective cyclooxygenase-2 inhibitors (Sc-58125 and Celecoxib) at various concentrations for 24 h. Morphological changes were observed under the phase microscopy and the growth suppression was detected by using MTT colorimetric assay. Apoptotic DNA fragments were observed by agarose gel electrophoresis, and the cell cycle and apoptotic rate were detected by flow cytometry (FCM) respectively. hep-2 cells became rounded and detached from the culture dish after being treated with Celecoxib for 24 h, however, they remained morphologically unchanged with Sc-58125. Sc-58125 could increase G 2 phase cells, whereas, Celecoxib rose G 1 phase cells. Both of the two effects were dose-dependent. Moreover, the hep-2 cells cultured with 50 μmol/L and 100 μmol/L Celecoxib showed obvious apoptosis, with the nuclear DNA of cells exhibiting characteristic DNA ladder. So Sc-58125 could inhibit the proliferation of hep-2 cells by altering the G 2 phase cells. However, Celecoxib had the same effect by changing the G 1 phase cells and inducing apoptosis at higher concentration.

关键词： Sc-58125 Celecoxib carcinoma of larynx hep-2 cell Inhibition proliferation

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