【作者】 秦艳芳；

【导师】 向军俭；

【作者基本信息】 暨南大学 ， 免疫学， 2002， 硕士

【摘要】 [目的]从细胞生物学的角度探讨细胞的恶性表型与bFGF的高表达的关系，为bFGF作为肿瘤诊断标记物提供依据，并为靶向bFGF的肿瘤治疗打下基础。 [主要方法]以SWO-38神经胶质瘤细胞系、ASTC-A-1肺癌细胞系、BCAP-37乳腺癌细胞、HepG-2肝癌细胞系、Hep-2喉癌细胞系为研究对象，将bFGF反义寡核苷酸导入肿瘤细胞，同时设正义寡核苷酸对照组，用免疫组化结合图象分析技术检测胞内bFGF的表达、ELISA法测定细胞上清bFGF、原位杂交结合图象分析技术检测bFGFmRNA，比较导入bFGF反义寡核苷酸前后的bFGF、bFGFmRNA的量的改变，以证实bFGF表达受抑制。然后用MTT法观察bFGF表达受抑制的肿瘤细胞生长的变化、半固体琼脂糖集落形成率的改变；选取SWO-38神经胶质瘤细胞系、BCAP-37乳腺癌细胞为对象，用免疫组化结合图象分析的技术检测FGFRl的表达变化。并用扫描电镜观察SWO-38神经胶质瘤细胞系细胞形态的变化。 [结果]bFGF表达抑制的5种肿瘤细胞表现出增殖率的下降、半固体琼脂糖集落形成能力的减弱。SWO-38神经胶质瘤细胞系、BCAP-37乳腺癌细胞的FGFR1表达也随之减少。SWO-38神经胶质瘤细胞细胞表面微绒毛形态和数量改变。 [结论]过量表达的内源性bFGF参与了多种细胞的恶性转化，在随后的肿瘤进展过程中也发挥着重要的作用。FGFR1与bFGF的恶性分泌环的放大效应是bFGF影响肿瘤细胞恶性表型的基础。不同类型的肿瘤细胞对bFGF的依赖性不同。bFGF、bFGFmRNA、FGFR1有望成为新的肿瘤诊断标志物；bFGF反义基因治 毕业论文：碱性成纤维细胞因子及受体1与肿瘤细胞恶性行为的关系 疗一种有前途的抗肿瘤手段。更多还原

【Abstract】 Aims:To examine the contribution of endogenous bFGF to the abnormal growth and phenotypic transformation to neoplastic tumors cells.Methods: In the present study ,the influnce of endogenous bFGFon human malignant brain glioma cells tumor SWO-38 cell line,lung carcinoma ASTC-A-1 cell line ,breast cancer BCAP-37 cell line,hepatocarcinoma HepG-2 cell line and laryngocarcinoma Hep-2 cell line growth in vitro was evaluted by down-regulating bFGFexpression using antisense oligonucleotide primers.The characteristic expression of bFGF and FGFR1 was examined by immunohistochemical methods and image analysis techniques.In situ hybridization and image analysis technique was used for bFGF mRNA expression.inaddition,extracellular bFGF was examined by ELISA .MTT spetrophotametry was used to examine the effect of bFGF antisense oligonucleotide primers on survival of the cells.Cloning efficency of the cells in soft agar was assessed after exposed to bFGF antisense oligonucleotide primers.Scanning electron microscopy was used to observe the change of SWO-38 cell surface. ResultrBFGF expression was by the addition of bFGF antisense oligonucleotide primers. The addition of bFGF-specific antisense oligonucleotides to the five types of tumor cells resulted in inhabition(from30% to 51%)in survival of the cells .bFGFantisense oligonucleotides also inhabited the colony formation in soft agar and changed Colony morphology also.These effects were dose-dependent and specific .In addition, FGFR1 expression was found reduced in responds tobFGF-specific antisense primers when we studied SWO-38 cell line and BCAP-37 cell line.There are some changes of SWO-38 cell surface when infected bFGF antisense oligonucleotide primers in electron microscopy.Conclusions:Endogenous bFGF can regulate the phenotypic malignacy in the humor tumor cells.The alterations in bFGF expression may play a role in the development and prosession of human neoplastic tumors.BFGFmay act as auntocrine regulators by interacting with FGFR1 on malignant tumors cells.BFGF ,FGFmRNA or FGFR1 lever in tumor tissue or tumor patientsis a useful giognotic marker. It is hopeful that bFGF antisense oligonucleotide primers may become a new thrapeutic drug for anti-tumor.更多还原