电压门控氯通道3可促进胶质瘤细胞的侵袭和迁移
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摘要
目的探讨电压门控氯通道3(CLC-3)在人脑胶质瘤细胞的表达以及在其侵袭和迁移中的作用。方法采用免疫组织化学(免疫组化)技术,检测30例临床确诊并有完整资料的人胶质瘤病理切片和6例非胶质瘤正常人脑标本中CLC-3蛋白的表达情况,再通过ShCLC-3干扰腺病毒下调U87MG细胞株CLC-3的表达,运用Transwell细胞迁移和侵袭实验检测胶质瘤细胞侵袭迁移情况,并进行统计学分析。结果 30例胶质瘤病理结果 WHO胶质瘤病理分级标准Ⅰ级6例、Ⅱ级9例、Ⅲ级7例、Ⅳ级8例;在高级别(Ⅲ～Ⅳ)胶质瘤细胞中染色呈明显棕黄色或棕褐色,在低级别胶质瘤(Ⅰ～Ⅱ)中染色呈淡黄色或棕黄色,在正常脑细胞中可见1例无着色,另外5例呈淡黄色;CLC-3蛋白的表达与性别、年龄无相关性(P均> 0.05)。Spearman秩相关分析结果显示CLC-3的表达与WHO胶质瘤病理分级呈正相关(rs=0.782,P <0.001)。通过ShCLC-3腺病毒感染胶质瘤细胞株U87MG后CLC-3表达降低,胶质瘤细胞的侵袭和迁移能力减弱(P均<0.05)。结论 CLC-3在胶质瘤细胞中高表达并与其病理分级呈正相关,CLC-3可促进胶质瘤的侵袭和迁移。
Objective To investigate the expression and effect of chloride channel-3(CLC-3) on the invasion and migration of human glioma cells. Methods Immunohistochemical staining was adopted to detect the expression of CLC-3 protein in the pathological sections of 30 glioma patients with complete data and brain specimens from 6 non-glioma normal subjects. The expression of CLC-3 in the U87 MG cell lines was down-regulated by ShCLC-3 interfering adenovirus. The invasion and migration of glioma cells were detected by Transwell invasion and migration assay. The detection results were statistically analyzed. Results According to the pathological grading criteria of glioma by WHO, 6 cases were classi?ed as grade Ⅰ, 9 cases of grade Ⅱ, 7 cases of grade(grade Ⅲ-Ⅳ), light yellow or yellow in the low-grade glioma(grade Ⅰ-Ⅱ). In the normal brain cells, CLC-3 was not stained in 1 case, and light yellow in the remaining 5 cases. The expression of CLC-3 protein was not signi?cantly correlated with gender or age(both P > 0.05). Spearman rank correlation analysis demonstrated that the expression of CLC-3 was positively correlated with WHO pathological grading of glioma(rs = 0.782, P <0.001). The expression of CLC-3 was signi?cantly down-regulated in U87 MG cells after the cells were infected by the ShCLC-3 adenovirus, and the invasion and migration abilities of glioma cells were signi?cantly reduced(both P < 0.05). Conclusions CLC-3 is highly expressed in the pathological specimens of glioma, which is positively correlated with pathological grading of glioma. CLC-3 can promote the invasion and migration of glioma cells.
Objective To investigate the expression and effect of chloride channel-3(CLC-3) on the invasion and migration of human glioma cells. Methods Immunohistochemical staining was adopted to detect the expression of CLC-3 protein in the pathological sections of 30 glioma patients with complete data and brain specimens from 6 non-glioma normal subjects. The expression of CLC-3 in the U87 MG cell lines was down-regulated by ShCLC-3 interfering adenovirus. The invasion and migration of glioma cells were detected by Transwell invasion and migration assay. The detection results were statistically analyzed. Results According to the pathological grading criteria of glioma by WHO, 6 cases were classi?ed as grade Ⅰ, 9 cases of grade Ⅱ, 7 cases of grade(grade Ⅲ-Ⅳ), light yellow or yellow in the low-grade glioma(grade Ⅰ-Ⅱ). In the normal brain cells, CLC-3 was not stained in 1 case, and light yellow in the remaining 5 cases. The expression of CLC-3 protein was not signi?cantly correlated with gender or age(both P > 0.05). Spearman rank correlation analysis demonstrated that the expression of CLC-3 was positively correlated with WHO pathological grading of glioma(rs = 0.782, P <0.001). The expression of CLC-3 was signi?cantly down-regulated in U87 MG cells after the cells were infected by the ShCLC-3 adenovirus, and the invasion and migration abilities of glioma cells were signi?cantly reduced(both P < 0.05). Conclusions CLC-3 is highly expressed in the pathological specimens of glioma, which is positively correlated with pathological grading of glioma. CLC-3 can promote the invasion and migration of glioma cells.
引文
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[15]Sciaccaluga M,Fioretti B,Catacuzzeno L,Pagani F,Bertollini C,Rosito M,Catalano M,D’Alessandro G,Santoro A,Cantore G,Ragozzino D,Castigli E,Franciolini F,Limatola C. CXCL12-induced glioblastoma cell migration requires intermediate conductance Ca2+-activated K+channel activity. Am J Physiol Cell Physiol,2010,299:C175-C184.
[16]Lin ZX. Glioma-related edema:new insight into molecular mechanisms and their clinical implications. Chin J Cancer,2013,32:49-52.
[17]Wu CX,Lin GS,Lin ZX,Zhang JD,Chen L,Liu SY,Tang WL,Qiu XX,Zhou CF. Peritumoral edema on magnetic resonance imaging predicts a poor clinical outcome in malignant glioma. Oncol Lett,2015,10:2769-2776.
[2]Farin A, Suzuki SO, Weiker M, Goldman JE, Bruce JN, Canoll P. Transplanted glioma cells migrate and proliferate on host brain vasculature:a dynamic analysis. Glia,2006,53(8):799-808.
[3]Fan F,Liu T,Wang X,Ren D,Liu H,Zhang P,Wang Z,Liu N,Li Q,Tu Y,Fu J. ClC-3 expression and its association with hyperglycemia induced HT22 hippocampal neuronal cell apoptosis. J Diabetes Res,2016,2016:2984380.
[4]Huang LY,He Q,Liang SJ,Su YX,Xiong LX,Wu QQ,Wu QY,Tao J,Wang JP,Tang YB,Lv XF,Liu J,Guan YY,Pang RP,Zhou JG. ClC-3 chloride channel/antiporter defect contributes to inflammatory bowel disease in humans and mice. Gut,2014,63(10):1587-1595.
[5]Zhang H,Li H,Liu E,Guang Y,Yang L,Mao J,Zhu L,Chen L,Wang L. The AQP-3 water channel and the ClC-3 chloride channel coordinate the hypotonicity-induced swelling volume in nasopharyngeal carcinoma cells. Int J Biochem Cell Biol,2014,57:96-107.
[6]Hong S, Bi M, Wang L, Kang Z, Ling L, Zhao C. CLC-3channels in cancer(review). Oncol Rep,2015,33(2):507-514.
[7]Pasantes-Morales H. Channels and volume changes in the life and death of the cell. Mol Pharmacol. 2016,90(3):358-370.
[8]Holm A,Magnusson KE,Vikstr?m E. Pseudomonas aeruginosa-3-oxo-dodecanoyl-homoserine lactone elicits changes in cell volume, morphology, and aqp9 characteristics in macrophages.Front Cell Infect Microbiol,2016,6:32.
[9]Parkerson KA, Sontheimer H. Contribution of chloride channels to volume regulation of cortical astrocytes. Am J Physiol Cell Physiol,2003,284(6):C1460-C1467.
[10]Sontheimer H. An unexpected role for ion channels in brain tumor metastasis. Exp Biol Med(Maywood),2008,233(7):779-791.
[11]Thompson EG, Sontheimer H. A role for ion channels in perivascular glioma invasion. Eur Biophys J,2016,45(7):635-648.
[12]Sforna L,Cenciarini M,Belia S,Michelucci A,Pessia M,Franciolini F,Catacuzzeno L. Hypoxia modulates the swellingactivated cl current in human glioblastoma cells:role in volume regulation and cell survival. J Cell Physiol,2016,232(1):91-100.
[13]Kunzelmann K. TMEM16, LRRC8A, bestrophin:chloride channels controlled by Ca2+and cell volume. Trends Biochem Sci,2015,40(9):535-543.
[14]Olsen ML, Schade S, Lyons SA,Amaral MD,Sontheimer H.Expression of voltage-gated chloride channels in human glioma cells. J Neurosci,2003,23:5572-5582.
[15]Sciaccaluga M,Fioretti B,Catacuzzeno L,Pagani F,Bertollini C,Rosito M,Catalano M,D’Alessandro G,Santoro A,Cantore G,Ragozzino D,Castigli E,Franciolini F,Limatola C. CXCL12-induced glioblastoma cell migration requires intermediate conductance Ca2+-activated K+channel activity. Am J Physiol Cell Physiol,2010,299:C175-C184.
[16]Lin ZX. Glioma-related edema:new insight into molecular mechanisms and their clinical implications. Chin J Cancer,2013,32:49-52.
[17]Wu CX,Lin GS,Lin ZX,Zhang JD,Chen L,Liu SY,Tang WL,Qiu XX,Zhou CF. Peritumoral edema on magnetic resonance imaging predicts a poor clinical outcome in malignant glioma. Oncol Lett,2015,10:2769-2776.