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High-grade Glioma Motility Reduced by Genetic Knockdown of KCC3

机译：KCC3基因敲低降低了高级胶质瘤运动性。

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Cell motility is dependent on a coordinated reorganization of the cytoskeleton, membrane recycling, and focal adhesion to the extracellular matrix. Each of these cellular processes involves re-distribution of cell water, which is facilitated by the transport of inorganic ions (with obligatory water movement). Scratch-wound healing assays of Wistar C6 glioblastoma cells demonstrated cell motility in advance of cell proliferation. Although bumetanide inhibition of Na-K-2Cl cotransport activity did not affect cell motility, treatment of glioma cells with furosemide to inhibit K-Cl cotransport activity prevented ～75% of wound closure in a reversible reaction. Genetic silencing of KCC3 with short hairpin interfering RNA reduced protein expression by 40 - 60%, Ksup+/sup influx by ～50%, and cell motility by ～50%. Appearance of KCC1 mRNA and KCC3 mRNA at 25 PCR cycles versus KCC4 mRNA at 35 PCR cycles, suggests more KCC1/KCC3 expression in both primary rat astrocytes and C6 glioma cells. Altogether, these experiments suggest that the presence/function of multiple isoforms of the Nasup+-/supindependent K-Cl cotransporter may have a role in glioma cell motility.
机译：细胞运动性取决于细胞骨架的协调重组，膜循环以及对细胞外基质的粘着。这些细胞过程中的每一个都涉及细胞水的重新分布，这通过无机离子的运输（强制水运动）而得以促进。 Wistar C6胶质母细胞瘤细胞的划伤愈合试验证明了细胞增殖之前的细胞运动性。尽管布美他尼对Na-K-2Cl共转运活性的抑制作用不影响细胞运动性，但应用呋塞米抑制K-Cl共转运活性的神经胶质瘤细胞在可逆反应中可防止约75％的伤口闭合。带有短发夹干扰RNA的KCC3的基因沉默使蛋白质表达降低40-60％，K + 流入减少约50％，而细胞运动减少约50％。与在35个PCR周期的KCC4 mRNA相比，在25个PCR周期的KCC1 mRNA和KCC3 mRNA的出现表明在原代大鼠星形胶质细胞和C6胶质瘤细胞中都有更多的KCC1 / KCC3表达。总而言之，这些实验表明，Na +-独立的K-Cl共转运蛋白的多种同工型的存在/功能可能在神经胶质瘤细胞运动中起作用。

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