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目的观察Flt-1表达降低对胶质瘤细胞株的生长侵袭性和迁移能力的生物学影响,探讨Flt-1对胶质瘤侵袭以及迁移过程的作用。方法 (1)体外培养大鼠神经胶质瘤C6细胞,利用腺病毒载体小RNA干扰技术,建立Flt-1表达降低的细胞株。(2)通过脑立体定向技术将Flt-1表达降低的胶质瘤C6细胞(实验组A)和未作特殊处理的C6细胞(对照组B)注射到SD大鼠右侧尾状核部,相同营养条件下饲养两组SD大鼠,借助磁共振平扫观察SD大鼠颅内肿瘤的生长侵袭情况。20d后处死SD大鼠,并冠状切开注射胶质瘤C6细胞部位的颅脑组织,制备石蜡切片,HE染色,计数切片中注射部位及周边的肿瘤卫星结节数量。(3)把Flt-1表达降低的胶质瘤C6细胞(实验组C)和未经特殊处理的C6细胞(对照组D)分别制造细胞划痕,记录6h,12h,18h两组划痕的宽度。结果实验组A切片标本中肿瘤卫星结节的数量(29.0±17.3)明显少于对照组B切片标本中的结节数量(84.0±20.5),Flt-1表达降低的胶质瘤C6细胞的生长侵袭性明显降低(P<0.05);实验组C 6h划痕宽度(547±45.8μm)、12h划痕宽度(433±42.2μm)、18h划痕宽度(268±43.7μm)分别明显大于对照组D中相应的6h划痕宽度(470±43.6μm)、12h划痕宽度(357±45.6μm)、18h划痕宽度(195±44.6μm),Flt-1表达降低的胶质瘤C6细胞株的迁移能力明显降低(P<0.05)。结论 Flt-1可能提高神经胶质瘤细胞的生长侵袭性和迁移能力,二者之间有着密切的关系。
Objective To investigate the biological effect of Flt-1 on glioma cell invasion and migration and to investigate the role of Flt-1 in glioma invasion and migration. Methods (1) C6 glioma cells were cultured in vitro, and the cell lines with reduced expression of Flt-1 were constructed by adenovirus vector small RNA interference. (2) Glioma C6 cells with reduced Flt-1 expression (experimental group A) and C6 cells without special treatment (control group B) were injected into the right caudate nucleus of SD rats by brain stereotactic technique. Two groups of SD rats were fed under the same nutritional conditions, and the growth and invasion of intracranial tumors in SD rats were observed by plain magnetic resonance imaging. After 20 days, the SD rats were sacrificed and the brain tissue of the C6 glioma site was excised by coronary angiography. Paraffin sections were prepared and stained with hematoxylin and eosin (HE). The number of tumor satellite nodules at the site of injection and peripheral tumor was counted. (3) Scratches were made in glioma C6 cells with reduced expression of Flt-1 (experimental group C) and C6 cells without special treatment (control group D), respectively. Scratches were recorded for 6h, 12h and 18h width. Results The number of tumor satellite nodules in experimental group A (29.0 ± 17.3) was significantly less than that in control group (84.0 ± 20.5), the expression of Flt-1 was decreased in glioma C6 cells (P <0.05). The widths of scratches at 6h (547 ± 45.8μm), scratches at 12h (433 ± 42.2μm) and scratches at 18h (268 ± 43.7μm) were significantly higher in the experimental group than those in the control group The corresponding 6h scratch width (470 ± 43.6μm), 12h scratch width (357 ± 45.6μm), 18h scratch width (195 ± 44.6μm) in D, and the glioma C6 cell line with reduced Flt-1 expression Migration ability was significantly reduced (P <0.05). Conclusion Flt-1 may increase the invasiveness and migration ability of glioma cells, and there is a close relationship between them.