哺乳动物的几种培养细胞的动粒蛋白的研究

STUDIES ON KINETOCHORE PROTEINS IN SEVERAL CULTURED MAMMALIAN CELLS

用来自人类硬皮病中一种称为ＣＲＥＳＴ综合征的自身免疫性疾病病人的抗动粒抗体血清（ＡＣＡ血清）和蛋白质免疫印迹技术，研究了来源于人（ＨＬ６０细胞，ＨｅＬａ细胞以及外周血淋巴细胞）和小鼠（ＮＩＨ３Ｔ３细胞，转化的ＬＡ９０细胞，ＴＣ３Ｈ１０ Ｔ１／２细胞）的６种培养细胞的动粒蛋白（ｋｉｎｅｔｏｃｈｏｒｅｐｒｏｔｅｉｎ）。用间接免疫荧光法显示此种ＡＣＡ血清对哺乳动物培养细胞的动粒（ｋｉｎｅｔｏ－ｃｈｏｒｅ）有特异的染色反应。此种血清在上述细胞中共识别出６种动粒蛋白，我们命名为ＡＡ１（１７ｋＤ）、ＡＡ２（５０ｋＤ）、ＡＡ３（６０ｋＤ）、ＡＡ４（７７ｋＤ）、ＡＡ５（８０ｋＤ）、ＡＡ６（１４０ｋＤ）。其中，ＡＡ１、ＡＡ５、ＡＡ６分别相当于文献报道的ＣＥＮＰ－Ａ、ＣＥＮＰ－Ｂ、ＣＥＮＰ－Ｃ。本研究结果表明，ＣＥＮＰ－Ｂ在上述细胞中分布最广，存在于ＨｅＬａ、ＨＬ６０、ＮＩＨ３Ｔ３、ＬＡ９０、ＴＣ３Ｈ１０ Ｔ１／２等５种细胞，其次为ＣＥＮＰ－Ｃ和ＣＥＮＰ－Ａ，ＣＥＮＰ－Ｃ存在于ＨＬ６０、ＮＩＨ３Ｔ３、ＬＡ９０、和ＴＣ３Ｈ１０ Ｔ１／２细胞，而ＣＥＮＰ－Ａ存在于ＨｅＬａ和淋巴细胞。此外还提示，癌细胞或转化细胞的动粒蛋白的种类较正?

Using ACA(anticentromere/kinetochore antibody)serum from a patient with CREST variant of scleroderma,the kinetochore proteins were studied in six kinds of cultured mammalian cells including cells derived from human(HeLa,HL60,and peripheral lymphocytes) and mouse (NIH3T3,transformed LA90,and transformed C3H10T1/2)by indirect immunofluorescence and immunoblotting methods. Indirect immunofluorescence showed that the ACA serum used is specific to the kinetochore /prekinetochore of the mammalian cells,It could recognize six types of kinetochore proteins in all the cultured cells mentioned above,i.e. ,AA1(CENP-A,17kD),AA2(50kD),AA3(66kD),AA4(77kD),AA5(CENP-B,80kD),AA6(CENP-C,140 kD).Furthermore,it was found that CENP-B existed in HeLa,HL60,NIH3T3, transformed LA90,and transformed C3H10T1/2,while CENP-C existed in HL60,NIH3T3,transformed LA90,and transformed C3H10T1/2, CENP-A existed in HeLa and lymphocytes.The results also suggested that the types of kinetochore proteins in cancer or transformed cells are more than those in normal ones.