摘要
目的探讨6A8α-甘露糖苷酶表达抑制对CNE-2L2细胞与层粘连蛋白的粘附及细胞表面片状伪足的影响。方法Western印迹检测6A8α-甘露糖苷酶的表达。置细胞于层粘连蛋白(laminin)包被的培养板孔中,37℃孵育1h后,用结晶紫染色细胞,用0.1mol/L枸橼酸钠/50%乙醇溶解结合到细胞中的染料。测定540nm处的吸光度值(AT),按R=AT/A100×100%计算粘附率(A100为100%粘附值)。扫描电镜观察细胞表面的片状伪足。结果6A8α-甘露糖苷酶表达抑制的2个克隆细胞株(AS1和AS2)对层粘连蛋白的相对粘附率分别为0.447±0.096与0.533±0.065,而6A8α-甘露糖苷酶表达正常的对照细胞(W、M和S)的相对粘附率分别为0.78±0.035、0.7±0.05和0.80±0.04。两者的差别有生物学意义(P<0.01)。6A8α-甘露糖苷酶表达正常的细胞表面片状伪足丰富,6A8α-甘露糖苷酶表达抑制的细胞表面的片状伪足基本上消失。结论6A8α-甘露糖苷酶表达抑制可导致人鼻咽癌细胞CNE-2L2与层粘连蛋白粘附减弱及片状伪足减少。
Objective To investigate the inhibitory effect of6A8α-manosidase expression on the adhesiveness of CNE-2L2cells to laminin and the lamellipodia on cell surface.Methods 6A8α-manosidase expression was detected by Western blotting.For assaying the adhesion of cells to laminin,cells were incubated in laminin-coated plate at37℃for1h,the adhered cells were stained with crystal purple dissolved in0.1mol/L Sodium Citrate/50%ethanol.Absorbance540nm was measured.Adhesion rate(R)was calculated according to formula R=A T /A 100 ×100%.Here A 100 represents100%adhesion.lamellipodia on cell surface was observed upon a scanning electron microscopy.Results The adhesion rate of two clones(AS1and AS2)with inhibition of6A8α-manosidase expression to laminin was0.447±0.096and0.533±0.065respectively.The adhesion rate of three controls with normal expression of6A8α-manosidase to laminin was0.78±0.035、0.7±0.05and0.80±0.04respectively.The difference was significant (P<0.01).CNE-2L2cells with normal expression of6A8α-manosidase was rich in lamellipodia on their surface.Lamellipodia nearly disappeared on the cells with inhibition of6A8α-manosidase expression.Conclusions Inhibition of6A8α-manosidase expression results in decrease of adhesion to laminin and reduction of lamellipodia of human nasopharyngeal carcinoma cell CNE-2L2.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2003年第3期316-319,共4页
Acta Academiae Medicinae Sinicae
基金
北京市科委项目(H020220020310)资助~~
