Protein expression and functional analysis of the FHIT gene in human tumor cells
Protein expression and functional analysis of the FHIT gene in human tumor cells
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DOI：
10.1093/jnci/90.6.426
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发表时间：
1998-03-18
期刊：
影响因子：
--
通讯作者：
Yeung, RS
中科院分区：
文献类型：
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作者：
Otterson, GA;Xiao, GH;Yeung, RS
背景：染色体3p14.2上的脆性组氨酸三联体(FHIT)基因已被认为是人类肿瘤中的候选抑癌基因，为了验证FHIT是否具有抑癌基因的功能特性，我们研究了FHIT蛋白在人癌细胞中的表达，并检测了FHIT抑制肿瘤细胞表型的能力。方法：通过免疫组织化学分析和免疫印迹方法确定肿瘤细胞的亚细胞定位和蛋白表达模式，在pFHIT检测不到的肿瘤细胞中，我们检测了重组pFHIT表达对肿瘤形态、生长速度、克隆形成以及体内肿瘤形成和形成的影响。结果：pFHIT是一种胞质17-kd多肽，在52株人癌细胞中有30株未检测到pFHIT的表达，但pFHIT的稳定过表达并未改变细胞形态、抑制集落形成或抑制细胞体外增殖，且过表达pFHIT不会引起细胞分裂周期动力学的改变，高表达重组pFHIT的肿瘤细胞的体内致瘤性与对照组和亲本细胞相当。结论：这些结果表明，pFHIT在人类癌细胞中的替代不会抑制肿瘤细胞的生长，该蛋白可能以不同于经典的肿瘤抑制范例的方式参与肿瘤的发生。
Background: The fragile histidine triad (FHIT) gene at chromosome 3p14.2 has been proposed to be a candidate tumor suppressor gene in human cancers, To test whether FHIT exhibits the functional properties of a tumor suppressor gene, we studied the expression of its protein (pFHIT) in human carcinoma cells and examined the ability of FHIT to inhibit the neoplastic phenotype of cancer cells, Methods: Subcellular localization and patterns of protein expression in tumor cells were determined by immunohistochemical analysis and immunoblotting with the use of polyclonal anti-pFHIT antisera, In tumor cells with undetectable pFHIT, we examined the effect of recombinant pFHIT expression on morphology, growth rate, colony formation, and in vivo tumor formation, Results: We demonstrated that pFHIT is a cytoplasmic 17-kd polypeptide whose expression could not be detected in 30 of 52 human carcinoma cell lines tested, We observed, however, that the stable overexpression of pFHIT did not alter cell morphology, inhibit colony formation, or inhibit cell proliferation in vitro, Furthermore, overexpression of pFHIT did not lead to altered cell cycle kinetics in dividing cells, The in vivo tumorigenicity of a tumor cell line that expressed high levels of recombinant pFHIT was equivalent to that of control transfectants and of parental cells, Conclusions: These results suggest that the replacement of pFHIT in human carcinoma cells does not suppress tumor cell growth and that this protein may be involved in tumorigenesis in ways that are distinct from the "classic" tumor suppressor paradigm.