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目的观察靶向存活素基因的特异性短发卡 RNA(shRNA)对人脑胶质母细胞瘤 U251细胞裸鼠体内肿瘤生长和血管形成的影响。方法将 U251细胞、稳定转染存活素基因 shRNA 真核表达载体 pWH1-SR 的 U251细胞(U251-SR 细胞)、稳定转染空载体 pWH1的 U251细胞(U251-P 细胞),分别接种于15只裸鼠背部皮下,每组5只,观测肿瘤生长情况。采用免疫组化 SABC 方法检测存活素、增殖细胞核抗原(PCNA)以及第八因子相关抗原(FⅧRAg)在各组肿瘤标本中的表达;采用TUNEL 方法检测凋亡细胞,分别计算各组肿瘤标本的增殖指数(PJ)、凋亡指数(AI)以及微血管密度(MVD)。结果与 U251、U251-P 组相比,U251-SR 组裸鼠肿瘤形成时间延迟,肿瘤生长缓慢,肿瘤体积及重量均明显减小(P 均<0.01);肿瘤标本存活素蛋白表达明显下调;PI 和 MVD 明显减少,AI 明显升高(P 均<0.01)。结论靶向存活素基因的 shRNA 能够在裸鼠体内明显抑制 U251细胞的肿瘤生长和血管形成。
Objective To observe the effect of specific short hairpin RNA (shRNA) targeting Survivin gene on tumor growth and angiogenesis in human glioblastoma U251 cells in nude mice. Methods U251 cells, U251 cells (U251-SR cells) stably transfected with Survivin gene shRNA eukaryotic expression vector pWH1-SR and U251 cells (U251-P cells) stably transfected with empty vector pWH1 were inoculated into 15 The nude mice were subcutaneously injected with 5 mice in each group to observe the growth of tumor. The expression of survivin, proliferating cell nuclear antigen (PCNA) and factor VIII-related antigen (FⅧRAg) were detected by immunohistochemical SABC method in each group. Apoptotic cells were detected by TUNEL method and the proliferation of tumor specimens Index (PJ), apoptosis index (AI) and microvessel density (MVD). Results Compared with U251 and U251-P group, the tumor formation time of U251-SR group was delayed, tumor growth was slow, tumor volume and weight were significantly reduced (all P <0.01), and the expression of Survivin protein in tumor tissue of U251- PI and MVD decreased significantly, AI increased significantly (all P <0.01). Conclusion Survivin targeting shRNA can significantly inhibit the tumor growth and angiogenesis of U251 cells in nude mice.