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bcl- 2是在多种恶性肿瘤中高表达的抗凋亡基因 ,它的高表达是肿瘤细胞对各种凋亡诱导剂不敏感的主要原因之一 ,因此 bcl- 2基因是反义寡核苷酸治疗恶性肿瘤的良好靶基因。 F95 1是与其他反义寡核苷酸序列不同的针对 bcl- 2基因 m RNA的反义硫代磷酸寡脱氧核苷酸 ,FNS18与 F95 1的核苷酸组成相同 ,但序列混杂的无义对照序列。 目的 研究 F95 1对人 Burkitt’s淋巴瘤 CA 4 6细胞生物学特性的影响及治疗 CA4 6细胞裸鼠移植瘤的药效学 ,同时探索 F95 1与小剂量阿糖胞苷 (Ara- c)治疗 CA 4 6细胞裸鼠移植瘤的药效学 ;研究 F95 1在 BAL B/c小鼠体内的药代动力学过程 ;在体外研究 CA 4 6细胞对 F95 1的摄取及 F95 1在 CA4 6细胞内的分布。 方法 (1)用锥虫蓝拒染法检测细胞生长情况 ;用MTT法检测细胞生长抑制率 ;用荧光定量 RT- PCR及流式细胞仪检测细胞 bcl- 2 m RNA和 Bcl- 2蛋白表达 ;用原位凋亡 TU NEL试剂盒和线粒体凋亡试剂盒检测早期凋亡细胞。(2 )制作 CA 4 6细胞裸鼠移植瘤模型并治疗 ;治疗过程中检测肿瘤体积变化 ;治疗结束后称量肿瘤重量 ;瘤块病理学检查、透射电镜观察 ;用荧光定量 RT- PCR检测瘤细胞内 bcl- 2基因 m RNA的水平 ;用 TU NEL试剂盒检测瘤组织内的凋亡细胞 ;用流式细胞仪检测瘤细?
bcl-2 is an anti-apoptotic gene that is overexpressed in a variety of malignant tumors. Its high expression is one of the main reasons that tumor cells are insensitive to various apoptosis inducing agents. Therefore, the bcl-2 gene is an antisense oligonucleotide Acid is a good target for the treatment of malignancies. F95 1 is an antisense phosphorothioate oligodeoxynucleotide directed against the bcl-2 gene m RNA that differs from other antisense oligonucleotides in that the nucleotide composition of FNS18 and F95 1 is the same but the sequence is miscellaneous Control sequence. Objective To investigate the effect of F95 1 on the biological characteristics of human Burkitt’s lymphoma CA 4 6 cells and the pharmacodynamics of CA 4 6 cell xenografts in nude mice. To explore the effect of F95 1 and small dose of Ara-c on CA 4 6 cell xenografts in nude mice; the pharmacokinetics of F95 1 in BALB / c mice was studied; the uptake of F95 1 by CA 4 6 cells and the uptake of F95 1 in CA 4 6 cells Distribution. Methods (1) Trypan blue exclusion assay was used to detect cell growth; cell growth inhibition rate was detected by MTT assay; bcl-2 mRNA and Bcl-2 protein expression were detected by real-time PCR and flow cytometry; Early apoptotic cells were detected by in situ apoptosis TU NEL kit and mitochondrial apoptosis kit. (2) The model of CA 4 6 cell xenografts in nude mice was made and treated. The change of tumor volume was measured during the course of treatment. The weight of tumor was weighed after the treatment. The pathological examination of the tumor and the transmission electron microscope were performed. Intracellular bcl-2 gene m RNA levels; TU NEL kit detection of apoptotic cells in tumor tissue; using flow cytometry to detect tumor size?