Flt-1对胶质瘤细胞侵袭及迁移的影响

Study on the Effect of Flt-1 on the Invasion and Metastasis in Glioma Cells

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 invasion and metastasis in down-regulated expression of Flt-1 glioma cells,and try to explore the effect of Flt-1 on the invasion and metastasis in glioma cells. Methods（1）Rats C6 glioma cells were cultivated in vitro. It got down-regulated expression of Flt-1 glioma cells by RNA interference.（2）Down-regulated expression of Flt-1 glioma cells（ experimental group A） and normal C6 cells（ control group B） were injected to the right side caudate nucleus of SD rats. Two groups of SD rats were fed in the same conditions. We detected the invasion and metastasis of intracranial tumor by MRI. Rats were killed after 20 days,and we made paraffin section of brain tissue.Paraffin section were processed by hematoxylin-eosin staining. It was analysed the effect of invasion by the number of tumor nodes.（3） Down-regulated expression of Flt-1 gliomacells（ experimental group C） and normal C6 cells（ control group D） were processed by making cells wound scratch. We kept a record of the scratch width of 6 hours/12 hours/18 hours in each group. Results The number of tumor nodes in paraffin section of the experimental group A（ 29. 0 ± 17. 3）were much smaller than the number of control group B（ 84. 0 ± 20. 5）. The invasion on glioma cells of down-regulated expression of Flt-1 was lower,the differences were statistically significant（ P 0. 05）. Scratch width of 6 hours（ 547. 0 ±45. 8μm）,12 hours（ 433. 0 ± 42. 2μm）,18 hours（ 268. 0 ± 43. 7μm） in experimental group C were much larger than in the control group D of 6 hours（ 470. 0 ± 43. 6μm）,12 hours（ 357. 0 ± 45. 6μm）,18 hours（ 195. 0 ± 44. 6μm）. Compared with control group,the metastasis on glioma cells of down-regulated expression of Flt-1 was lower,the differences were statistically significant（ P 0. 05）. Conclusion Flt-1 may play an crucial role in promoting the invasiveness and metastasis of glioma cells,and it is positively correlated with the invasion of glioma cells.