不同转移潜能鼻咽癌细胞钙电流特征与细胞迁徙能力的相关性  

Correlation of Ca^(2+) Current Features of Nasopharyngeal Carcinoma Cells with Different Metastatic Potentiality to Their Moving Abilities

作　　者：刘书静[1] 田道法[1] 何迎春[1] 曾亮[2] 黄辰[3] 刘宇勤[1] 

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410007 [2]湖南省肿瘤医院病理科,湖南长沙410018 [3]西安交通大学医学院中心实验室,陕西西安710061

出　　处：《癌症》2007年第5期473-479,共7页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30271678;No.30572408);国家研究生教育与学位中心基金项目(No.2003-46-08);国家中医药管理局中医药科研基金项目(No.02-03JP29);湖南省"十五;十一五"重点学科建设项目(No.2001-2010)~~

摘　　要：背景与目的:肿瘤转移活性与瘤细胞以迁徙运动潜能为代表的生物力学特性有关,而其迁移运动的生物力学机制又受制于细胞内的钙离子活动和钙电流特征。本文探讨不同转移潜能鼻咽癌单克隆亚系细胞钙电流特征,及与细胞迁徙运动潜能的相关性。方法:人鼻咽癌高转移潜能细胞株5-8F及低转移潜能细胞株6-10B分别与10%含药血清(由黄芪、党参、白花蛇舌草等组成)共同培养,MTT法检测含药血清对细胞增殖的影响,Western blot检测nm23-H1蛋白表达水平。利用全细胞膜片钳制方法观察细胞钙电流特征(药物作用20min),划痕实验观察细胞迁移能力(药物作用24h),比较分析其相关性。结果:6-10B细胞的nm23-H1表达水平(2.9±0.4)较5-8F细胞(2.3±0.21)高(P<0.05)。5-8F细胞内钙释放激活的钙电流ICRAC为(-1.39±0.36)nA,与6-10B细胞ICRAC[(-0.66±0.40)nA]相比差异有统计学意义(P<0.05)。通过划痕区的5-8F细胞数(350±3)也显著高于6-10B细胞(246±1)(P<0.05)。应用含药血清干预后,5-8F和6-10B细胞增殖活性差异无统计学意义(P>0.05),5-8F细胞的nm23-H1表达水平(3.9±0.1)明显高于6-10B细胞(1.0±0.1)(P<0.05)。干预后,5-8F细胞ICRAC降至(-1.27±0.35)nA,平均抑制幅度为(1.90±0.47)%;6-10B细胞则降至(-0.37±0.23)nA,平均抑制幅度为(0.46±0.12)%,两者差异有统计学意义(P<0.05),并出现与钙电流变化特点平行的细胞迁移能力变化,通过划痕线的5-8F细胞数明显减少(94±6),而6-10B细胞则受影响甚小(229±6,P<0.05)。干预前后5-8F细胞迁移能力变化具有显著性(P<0.05),6-10B细胞则无显著性差异(P>0.05)。结论:不同转移潜能鼻咽癌细胞钙电流特征及细胞迁移能力以及nm23-H1表达存在明显差异。应用含药血清干预后,高转移潜能细胞5-8F出现了与细胞钙电流ICRAC降低幅度相平行的细胞迁移能力抑制,药物血清可增加5-8F细胞的nm23-H1活性。BACKGROUND ＆ OBJECTIVE： The metastatic potentiality of malignancies is closely associated with their biological dynamic properties, which are affected by intracellular Ca^2± current activity, This study was to investigate the correlation of Ca^2± current features of sub-clonal nasopharyngeal carcinoma （NPC） cell lines 5-8F and 6-10B with different metastatic potentiality to their moving abilities, METHODS： 5-8F cells, with higher metastatic potentiality, and 6-10B cells, with lower metastatic potentiality, were cultured with herbal medicine-containing serum, which holds significant metastasis-inhibiting effect on tumor cells. Cell proliferation was assessed by MTT assay, The expression of nm23-H1 was detected by Western blot. Intracellular Ca^2± current features were detected with patch clamp technique in a whole cell recording way, while cell moving ability was determined by streak culturing assay, RESULTS： The expression of nm23-H1 was significantly lower in 5-8F cells than in 6-10B cells （2.3±0.2 vs. 2.9±0.4）. The Ca^2± release-activated Ca^2± influx current （ICRAC） was significantly lower in 5-8F cells than in 6-10B cells [（-1.39±0.36） nA vs. （-0.66±0.40） nA, P 〈 0.05]. The number of cells moved across the streak was significantly higher in 5-8F cells than in 6- 10B cells （350±3 vs. 246±1, P〈 0.05）. When cultured with herbal medicine-containing serum, no significant difference in proliferation was found between 5-8F cells and 6-10B cells; the expression of nm23-H1 was significantly higher in 5-8F cells than in 6-10B cells （3.9±0.1 vs.1.0±0.1 ,P〈0.05）; the ICRAC was decreased to （-1.27±0.35） nA in 5-8F cells and decreased to （-0.37±0.23） nA in 6-10B cell, and the inhibition rate was significantly higher in 5-8F cells than in 6-10B cells [ （1.90±0.47）% vs. （0.46±0.12）%, P 〈 0.05]; the number of cells moved across the streak was significantly lower in 5-8F cells than in 6-10B cells （94±6 vs. 229±6, P 〈 0.05）. CONCLUSIONS： The

关 键 词：鼻咽肿瘤 单克隆亚系细胞 钙电流特征 转移潜能 

分 类 号：R739.63[医药卫生—肿瘤]