水通道蛋白4与人星形胶质细胞瘤迁移的相关性

Role of Aquaporin-4 in Migration of Human Astrocytoma

目的探讨水通道蛋白4(aquaporin-4,AQP4)和星形胶质细胞瘤恶性程度和迁移能力的相关性。方法收集临床组织标本,培养获得正常星形胶质细胞、低级别星形细胞瘤和高级别星形细胞瘤细胞,荧光定量PCR法检测细胞中AQP4基因相对表达量,免疫印迹法检测AQP4蛋白水平,Transwell法检测细胞迁移能力。结果荧光定量PCR结果提示低级别组和高级别组内AQP4基因相对于对照组的扩增倍数分别为(1.65±0.052)和(2.04±0.196),差异有统计学意义(P<0.05);免疫印迹检测结果提示AQP4/β-actin灰度值比,对照组(0.103±0.012),低级别组(0.185±0.100),高级别组(0.248±0.028),各组间差异有统计学意义(P<0.05),AQP4蛋白表达水平随细胞恶性程度的增高而增高(P<0.05);Transwell法检测结果提示各组细胞脱色液吸光度值分别为对照组(0.408±0.010),低级别组(0.590±0.033),高级别组(0.986±0.026),差异有统计学意义(P<0.05),各组细胞迁移随AQP4表达的增高,细胞迁移能力也增强(P<0.05)。结论星形胶质细胞瘤中水通道蛋白4的表达随恶性程度的增高而增高,随AQP4表达的上升,星形胶质细胞瘤的迁移能力也增强。

Objective To investigate the effect of aquaporin-4(AQP4) on the severity and migration of astrocytoma. Methods Clinical tissue specimens were collected. Normal astrocyte, low-grade astrocytoma cells and high-grade astrocytoma cells were cultured from these specimens. Aquaporin-4 gene relative amount was detected by realtime fluorescence quantitative PCR. And the expression of aquaporin-4 protein was detected by Western blot. The migration of cells was measured with Transwell assay. Results The amplification of AQP4 gene in low grade group and high grade group were(1.65±0.052) and(2.04±0.196) separately, with statistically significant difference(P<0.05). Western blot results suggested that AQP4/β-actin grey value were(0.103±0.012) in control group,(0.185±0.100) in low grade group,(0.248±0.028) in high grade group, with significant difference(P<0.05); the level of aquaporin-4 protein was increased along with the growing of cell malignant degree(P<0.05). Transwell assay results suggested that the absorbance of cell destaining solution were(0.408±0.010) in control group,(0.590±0.033) in low grade group,(0.986±0.026) in high grade group, with significant difference(P<0.05). The cell migration ability was increased along the increasing of AQP4 expression(P<0.05). Conclusion Aquaporin-4 is contained in astrocytoma. With the increasing of malignant degree of astrocytoma, the level of AQP4 is also increased. The migration ability of astrocytoma is increased along with the increasing level of AQP4.