Role of Aquaporin-4 in Migration of Human Astrocytoma
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摘要: 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。Abstract: 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.
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Key words:
- Aquaporin-4(AQP4) /
- Astrocytoma /
- Cell migration
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[1] Hu J, Verkman AS. Increased migration and metastatic potential of tumor cells expressing aquaporin water channels[J]. FASEB J, 20 06, 20(11): 1892-4. [2] Wolburg H, Noell S, Fallier-Becker P, et al .The disturbed bloodbrain barrier in human glioblastoma[J]. Mol Aspects Med, 2012, 33 (5-6): 579-89. [3] Wang Y, Guan SS, Zhao G, et al. Expressions of aquaporin-4,matrix metallo-proteinase-2 and matrix metallo-proteinase-14 in peritumor edematous zone of glioma and clinical implications[J]. Zhonghua Yi Xue Za Zhi, 2014, 94(29): 2290-2. [王阳, 关树森, 赵岗, 等. 水通道 蛋白4、基质金属蛋白酶-2, -14在胶质瘤瘤周水肿组织中的表达 及其临床意义[J]. 中华医学杂志, 2014, 94(29): 2290-2.] [4] Mobasheri A, Marples D. Expression of the AQP-1 water channel in normal human tissues: a semiquantitative study using tissue microarray technology[J]. Am J Physiol Cell Physio, 2004, 286: C529-37. [5] Nielsen S, Smith BL, Christensen EI, et al. Distribution of the aquaporin CHIP in secretory and resorptive epithelia and capillary endothelia[J]. Proc Natl Acad Sci USA, 1993, 90(15): 7275-9. [6] Ding T, Ma Y, Li W, et al. Role of aquaporin-4 in the regulation of migration and invasion of human glioma cells[J]. Int J Oncol, 20 11, 38(6): 1521-31. [7] Oshio K, Binder DK, Liang Y, et al. Expression of the aquaporin- 1 water channel in human glial tumors[J]. Neurosurgery, 2005, 56 (2) : 375-81. [8] Longatti P, Basaldella L, Orvieto E, et al. Aquaporin 1 expression in cystic hemangioblastomas[J]. Neurosci Lett, 2006, 392(3): 17 8-80. [9] Saadoun S, Papadopoulos MC, Hara-Chikuma M, et al. Impairment of angiogenesis and cell migration by targeted aquaporin- 1 gene disruption[J]. Nature, 2005, 434(7034): 786-92. [10] Zhang SB, Dai DW, Dai YM. Advances of aquaporin-4 research[J]. Zhongguo Lin Chuang Shen Jing Ke Xue, 2013, 21(4): 451-5. [张 士保, 代大伟, 代亚美. 水通道蛋白-4的研究进展[J]. 中国临床 神经科学, 2013, 21(4): 451-5.] [11] Schwab A. Function and spatial distribution of ion channels and transporters in cell migration[J]. Am J Physiol Renal Physiol, 20 01, 280 (5): F739-47.
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