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［摘
要］ 目的
观察抗角蛋白抗体对角蛋白酪氨酸磷酸化的影响。
方法
以鼠抗人角蛋白的IgG单克隆抗体作用于培养中的Tca8113，以抗乙肝病毒表面抗原抗体做阴性对照，同时设置阳性对照和空白对照。细胞固定、包埋后以胶体金标记的羊抗鼠IgG抗体染色，用透射电镜观察。提取抗体作用过Tca8113细胞角蛋白，以抗乙肝病毒表面抗原抗体作用的细胞做阴性对照。不同分子量的角蛋白经SDSPAGE分离后转膜，用抗酪氨酸磷酸化抗体作用、显色。
结果
抗角蛋白抗体作用的Tca8133细胞胞质的中间丝有胶体金颗粒，与阳性对照组类似。阴性对照组及空白对照均未见胶体金颗粒。转膜后染色，在分子量大约58KD处出现阳性条带。阴性对照未出现条带。
结论
抗角蛋白抗体结合于胞浆中间丝，58KD角蛋白发生酪氨酸磷酸化。
［关键词］
抗角蛋白抗体；角蛋白；酪氨酸磷酸化
［中图分类号］
R 392.11 ［文献标识码］ A ［文章编号］
1001-7089(2005)09-0-0
Effects
of Antikeratin Antibody on Keratin of Live
Human Tca Cell line
WU
Caixia，LIU Yufeng，
LI Wei，et al
〖WT5”BX〗(Center
of Dermatology of Chinese PLA Xijing Hospital，
Fourth Military Medical University， Xi’an
710032，China)
Abstract:
Objective To study effects of
antikeratin antibody on keratin of live Human Tca cells.
Methods
Human Tca cells were primarily cultured in vitro
with antikeratin antibodies，and
some other Tca cells cultured with monoclonal antibodies to HbsAg served
as the negative group.Meanwhile the positive group and the blank group
were also established.All the cells were fixed and embedded before
stained with immunogold labeled antimouse
antibodies.Then the cells were observed under electron
microscope.Extract keratin from Tca cells cultured with antikeratin
antibodies.Keratin extracted from Tca cells with HbsAgs served as the
negative group.Keratin of different molecular were separated by SDS-PAGE
and transfered to nitrocellulose membrane(NC).And then keratin on NC
dyed after reacted with Phosphotyrosine Ab.
Results
The immunogold staining appeared on the keratin
intermediates of the cells cultured with antikeratin
antibodies.These phenomena were similar to what were seen in the
positive group.While immunogold staining were not seen in cells of the
nagetive and blank groups keratin of about 58KD on NC appeared after
transferring and dying.
Conclusion
Antikeratin antibodies
located in keratin intermediates of live Tca cells.The tyrosine
phosphorylation of 58KD keratin occurs.
Key
words： Antikeratin
monoclonal antibody；Keratin；Tyrosine
phosphorylation
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