[ABSTRACT]OBJECTIVETo study the mRNA expression of glucocorticoid receptor(GR) in peripheral blood mononuclear cells in patients with obstructive sleep apnea hypopnea syndrome(OSAHS) and its clinical significance. METHODSReal-time fluorescent quantitative PCR was used to detect GRα mRNA and GRβ mRNA exprssion in PBMC of 30 male patients with moderate to severe OSAHS and 27 healthy male subjects. The relationships between the expression of glucocorticoid receptor mRNA and the apnea hypopnea index, body mass index, neck circumference, waist circumference, the lowest oxygen saturation, the average oxygen saturation, the Epworth sleepiness scale(ESS) score, fasting blood glucose, heart rate and blood pressure were analyzed.RESULTSCompared with the control group, the expression level of GRα mRNA was lower in the OSAHS group(t=2.25,P<0.05) and the expression level of GRβmRNA had no difference between the two groups(t=1.43, P>0.05). We had not found the significant correlation between the expression of GRα mRNA and the clinical parameters above in OSAHS patients.CONCLUSIONThe expression of GRα mRNA in PBMC in moderate to severe OSAHS male patients have a downward trend compared with healthy group, but the mechanism remains unclear.

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae ; Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; Transfection

OBJECTIVETo study the effect and mechanism of tetrandrine (Tet) on enhancing radiosensitivity of human nasopharyngeal carcinoma cell lines in vitro.

METHODSCNE1 and CNE2 were exposed to radiation with or without Tet, the DNA damage of the cells were evaluated by neutral comet electrophoresis, and cell cycle and apoptosis were analyzed by flow cytometry.

RESULTSThe mean tail movements (TM) of CNE1 treated with radiation or radiation plus Tet were (7.13 ± 3.70) (X(-) ± s) and (13.61 ± 5.45), respectively (t = 2.784, P < 0.05), and TM of CNE2 treated with radiation or radiation plus Tet were (11.52 ± 4.04) and (18.85 ± 6.18), respectively (t = 3.089, P < 0.05). With the exposure to radiation or radiation plus Tet, the percentages of CNE1 in G2 phases were (42.62 ± 2.07)% and (17.02 ± 1.87)%, respectively (t = 23.173, P < 0.01), and the percentages of CNE2 in G2 phases were (34.82 ± 2.74)% and (19.64 ± 4.82)%, respectively(t = 16.500, P < 0.01). There was no significant difference in the apoptosis rates between the cells treated with radiation or radiation plus Tet regardless of CNE1 (17.24 ± 0.99)% vs (19.11 ± 1.24)%, and CNE2 (16.68 ± 0.27)% vs (18.51 ± 2.41)% (P > 0.05).

CONCLUSIONSTet can enhance radiosensitivity of human nasopharyngeal carcinoma cell lines. The mechanism could be related to abrogation of radiation-induced G2/M arrest and reduction of double-strand break repair capacity.

Apoptosis ; drug effects ; Benzylisoquinolines ; administration & dosage ; pharmacology ; toxicity ; Carcinoma ; Cell Line, Tumor ; DNA Repair ; Humans ; Nasopharyngeal Neoplasms ; Neoplasm Metastasis ; Radiation Tolerance ; drug effects

OBJECTIVETo explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.

METHODSA lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells. The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot, respectively. Cell proliferation was evaluated by CCK-8 assay, cell cycle and apoptosis were tested by flow cytometry, and cell invasion was detected by transwell invasion assay.

RESULTSThe relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P < 0.001). Western blot analysis proved that the relative expression of EGFL7 protein in NC group, Lenti-NC group and Lenti-EGFL7 group was (0.39 ± 0.12),(0.36 ± 0.14) and (0.07 ± 0.04), respectively. EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P < 0.001), which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells. The proliferation of Hep-2 cells was significantly inhibited after transfection (P < 0.01). Compared with the NC group and Lenti-NC group, the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P < 0.01), and the proportion in G1 phase was significantly decreased (P < 0.05). Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) % was significantly increased in NC group (6.09 ± 3.28) % and Lenti-NC group (9.86 ± 2.13) %. In Transwell invision assay, the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P < 0.01).

CONCLUSIONThe proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.

Apoptosis ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Cell Proliferation ; EGF Family of Proteins ; Gene Silencing ; Genetic Vectors ; Hep G2 Cells ; Humans ; Laryngeal Neoplasms ; metabolism ; virology ; Lentivirus ; RNA, Messenger ; RNA, Small Interfering ; Transfection

[摘 要] 目的：探讨肿瘤坏死因子受体相关蛋白1（TRAP1）在结肠癌组织和细胞中的表达及其与临床病理特征和患者预后的关系和相关分子机制。方法：通过TCGA和GEO数据全面分析TRAP1在结肠癌中的表达及其与临床病理特征和患者预后的关系，选取2020年10月至2021年03月间在山西医科大学第一医院手术切除的10例结肠癌组织及相应癌旁组织标本，用IHC染色法检测中国人结肠癌组织中TRAP1的表达进行验证，运行R包（survival和survminer）进行Kaplan-Meier生存分析；在线分析TRAP1蛋白的信号肽及穿膜结构域，通过基因富集分析软件进行GO分析和KEGG分析。培养结肠癌SW480和SW620细胞，将si-NC和si-TRAP1转染结肠癌细胞，实验分为空白对照组、si-NC组和si-TRAP1组，采用qPCR法检测转染后各组结肠癌细胞中TRAP1的表达，FCM检测转染后各组细胞的细胞周期和凋亡情况。结果：与癌旁组织比较，TRAP1在结肠癌组织中呈高表达（P<0.01），TRAP1表达水平与淋巴结转移有关联（P<0.05），TRAP1高表达组结肠癌患者5年OS率较低（P<0.05）。TRAP1蛋白属于细胞质蛋白，功能富集结果显示TRAP1及其相关分子与细胞周期、核糖体生物发生等信号通路有关（均P<0.01），TRAP1高表达组的结肠癌代谢重编程基因簇和线粒体蛋白输入基因簇水平升高（均P<0.01）。敲减TRAP1后，结肠癌细胞周期阻滞于G1期，细胞凋亡水平显著升高（均P<0.01）。结论：TRAP1在结肠癌组织中呈高表达，且与患者淋巴结转移和低OS率相关联，敲减TRAP1可阻滞结肠癌细胞周期并促进其凋亡。