表皮生长因受体拮抗剂在恶性脑胶质瘤细胞的抑制作用研究

Gliomas is account for about 45% of intracranial, in which the prognosis of malignant brain glioma with high rate of disability is poor. In the development process of glioma, there are many genetic and epigenetic changes. This brings difficulties to the treatment while these changes also give treatment to bring the opportunity.Therefore, the purpose of this study is to explore the effects of epidermal growth factor receptor antagonist on the proliferation of malignant glioma cells and its mechanism of action, which will find a more effective drugs for clinical glioma target for gene therapy. It has been successful to cultured primary glioma cells. In vitro experiments were performed to test synthesized drug, which demonstrated that in vitro on malignant glioma cells the drug has obvious inhibition. The inhibition was significantly higher than that of temozolomide group.The next, through the establishment of glioma model in nude mice, in vivo experiments were carried out using the EGFR antagonist with good effects in vitro, observing the growth of tumor with imaging; calculating the inhibition rate of tumor; testing the inhibition rate of drug on cell and the concentration of blood drug; getting the rate of blood brain barrier; using flow cytometry to analysis the effection of cell cycle and apoptosis after drug action on cells; detecting the expression of related protein with Immunohistochemistry and Western bloting. These would lay the foundation for the future clinical trial study and provide experimental data for authentic.

脑胶质瘤约占颅内肿瘤的45%，其中恶性脑胶质瘤预后差，致残率高。而在胶质瘤的发展过程中伴随着许多基因学和表观遗传学的变化，给治疗带来困难，同时这些改变也给治疗带来机会。因此，探寻表皮生长因受体拮抗剂对恶性胶质瘤细胞增殖的影响及其作用机制，为临床胶质瘤的基因靶向治疗寻找一种更有效的药物是本课题的目的。已成功培养原代胶质瘤细胞，并对已合成的试验药物进行体外实验，证明在体外对恶性胶质瘤细胞有明显抑制作用，其抑制作用明显高于替莫唑胺组。接下来将通过建立裸鼠胶质瘤模型，用体外实验中效果好的EGFR拮抗剂进行体内实验，影像学观察肿瘤的生长情况，计算瘤抑制率；检测药物对细胞的抑制率和和血药浓度; 血脑屏障的通过率；流式细胞仪分析药物作用于细胞后细胞增殖周期和细胞凋亡情况，免疫组织化学及Western bloting 检测相关蛋白表达。为以后临床试验研究打下基础、提供真实可信的实验数据。

脑胶质瘤约占颅内肿瘤的45%，其中恶性脑胶质瘤预后差，致残率高。而在胶质瘤的发展过程中伴随着许多基因学和表观遗传学的变化，给治疗带来困难，同时这些改变也给治疗带来机会。因此，探寻表皮生长因受体拮抗剂对恶性胶质瘤细胞增殖的影响及其作用机制，为临床胶质瘤的基因靶向治疗寻找一种更有效的药物是本课题的目的。已成功培养原代胶质瘤细胞，并对已合成的表皮生长因子受体拮抗剂LPY-5、LPY-9、吉非替尼以及替莫唑胺的体外研究，我们发现LPY-5和LPY-9对人脑胶质瘤细胞株U251具有明显增殖抑制及促进凋亡的作用，能够很好的抑制人脑胶质瘤细胞的迁移、侵袭，且作用效果优于吉非替尼、替莫唑胺。同时进行了人脑脑恶性胶质瘤的裸鼠皮下模型制作改良、原位模型制造尝试。探索并发现了采集大鼠脑脊液（血脑屏障相关研究中的重要样本）的新方法并对LPY-5、吉非替尼、替莫唑胺通过血脑屏障的能力进行了动物体内实验。进行了对长链非编码RNA SNHG12在脑胶质瘤中作用机制的探索，发现长链非编码RNA SNHG12在人脑胶质瘤组织及细胞系中高表达，且与肿瘤肿瘤的WHO分级以及患者预后显著相关。长链非编码RNA SNHG12参与人脑胶质瘤细胞株增殖、凋亡、迁移、转移等多个过程的的发展。长链非编码RNA SNHG12在细胞核和细胞质中均有表达，其可能通过竞争性结合miR-101-3p对FOXP1发挥调控作用，进而促进人脑胶质瘤的发生发展，为以后临床试验研究打下基础、提供真实可信的实验数据。