【作者】 李元伟；

【导师】 孙丰源；

【作者基本信息】 天津医科大学 ， 眼科学， 2010， 博士

【摘要】 目的1检测人眼眶横纹肌肉瘤(rhabdomyosarcoma,RMS)组织中legumain的表达,观察legumain的表达与RMS生物学行为之间的关系。并对横纹肌肉瘤患者的多个临床指标与患者预后进行多因素分析,对影响生存率的独立因素进行分析讨论。2检测经不同浓度的转化生长因子β(transforming growth factorβ,TGF-β)刺激之后人胚胎型RMS细胞系RD细胞中legumain及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达的变化,探讨legumain在RD细胞中的作用,为肿瘤的治疗提供新的思路和策略。方法1应用免疫组化SP法检测56例人眼眶RMS患者手术标本中legumain的表达,并引入年龄、性别、组织学类型、肿瘤大小、是否累及眼睑等因素进行统计学分析,观察各因素与患者预后的关系。2首先利用细胞免疫荧光法检测1egumain在RD细胞中的表达情况,并使用不同浓度的TGF-β对RD细胞进行刺激,通过RT-PCR以及Western-blot技术检测刺激前后legumain及α-SMA在RD细胞中表达的变化。结果1 legumain在56例RMS组织中有25例高表达,并且其表达高低、肿瘤大小、组织学类型、是否累及眼睑等因素是影响横纹肌肉瘤患者生存率的独立临床因素。2通过细胞免疫荧光法检测到RD细胞中存在legumain的表达,并且legumain及α-SMA的表达在2.5ng／ml的TGF-β刺激后较正常对照组明显升高,而5ng／ml刺激后RD细胞中legumain及α-SMA的表达较正常对照组明显降低。结论1 legumain在RMS组织中具有较高的表达,检测人眼眶RMS组织中legumain的表达有望成为判断肿瘤恶性程度以及患者预后的生物学指标,并为以后临床治疗眼眶RMS提供实验基础。2 Legumian低表达、肿瘤≦5cm、胚胎型RMS、肿瘤未累及眼睑的病人均具有较高生存率。而患者的年龄、性别因素与患者的预后无关。3低浓度的TGF-p刺激提高了RD细胞中legumain及α-SMA的表达,而高浓度的TGF-β刺激降低了RD细胞中legumain及α-SMA的表达,由于低浓度的TGF-β可以促进RD细胞的生长,而高浓度的TGF-β可以通过某种途径抑制RD细胞的生长,因此,legumain可能通过某种途径参与了TGF-β调控RD细胞生长的环节,从而增加了肿瘤细胞的侵袭性。更多还原

【Abstract】 Objective1 To investigate the expression of legumain in human orbital rhabdomyosarcoma as well as their relation with biological behaviour of human orbital rhabdomyosarcoma. And analyse the independent clinical factors that affected the survival of patients with rhabdomyosarcoma.2 To detect the expression of legumain and a-SMA in RD cell line stimulated with different concentration of TGF-βand invest the effect of legumain in RD cell line in order to provide new stratagies in tumor treatment.Methods1 The expression of legumain was evaluated in the 56 operative samples of human orbital rhabdomyosarcoma by immunohistochemistry and study the relationship between legumain expression and clinical factors including age, sex, histological type, tumor size, eyelid involved or not and prognosis of patients.2 The expression of legumain was detected by immunofluorescence. The expression of legumain and a-SMA in RD cell line stimulated with different concentration of TGF-βwere detected through RT-PCR and Western-blot.Results1 High expression of legumain were detected in 25 samples of human orbital rhabdomyosarcoma. The expression levels of legumain and the independent clinical factors such as histological type, tumor size, eyelid involved could affect survival of patients with rhbadomyosarcoma.2 The expression level of legumain and a-SMA in RD cell line stimulated with 2.5ng/ml of TGF-βwere higher than that of normal control group while stimulated with 5ng/ml the expression level of legumain and a-SMA in RD cell line were lower than that of normal control group.Conlusion1 The expression of legumain in human orbital rhabdomyosarcoma were high and detecting the expression of legumain may be become biological index for malignancy and prognosis factors of human orbital rhabdomyosarcoma. 2 Patients with low level of legumain exprssion, tumor size≦5cm, embryonal RMS, without eyelid involved had better prognosis.3 The expression level of legumain and a-SMA in RD cell line stimulated with 2.5ng/ml of TGF-βincreased while down regulated in RD cell line stimulated with 5ng/ml of TGF-β. Different concentration of TGF-βhave different effects on prolifaration of RD cell line, legumain may play important roles in the process of RD proliferation and increase the invasion activity of RD cells.更多还原