Restoration of fragile histidine triad (FHIT) expression induces apoptosis and suppresses tumorigenicity in lung and cervical cancer cell lines
Restoration of fragile histidine triad (FHIT) expression induces apoptosis and suppresses tumorigenicity in lung and cervical cancer cell lines
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DOI：
10.1073/pnas.062030799
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发表时间：
2002-03-19
影响因子：
11.1
通讯作者：
Sozzi, G
中科院分区：
文献类型：
--
作者：
Roz, L;Gramegna, M;Sozzi, G
Fhit蛋白表达的缺失通常与许多人类上皮癌的发展有关，包括肺癌和宫颈癌。然而，在源自这些肿瘤的细胞系中恢复Fhit的表达却产生了相互矛盾的结果，这促使人们需要仔细评估Fhit的抑癌潜力。在本研究中，我们研究了重新引入Fhit对7种肺癌和3种宫颈癌细胞系的影响。为了实现高效的基因转移和高水平的转基因表达，我们使用腺病毒载体转导FHIT基因。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法和碘化丙啶染色法评价细胞凋亡的诱导作用。Western blot检测caspase的激活情况，并对转染细胞在裸鼠体内的致瘤潜力进行评估。恢复Fhit表达可诱导所有Fhit阴性细胞系凋亡，其中肺癌细胞系中Calu-1、H460和A549最易感，宫颈癌细胞系中SiHa细胞最易感。caspase-8的激活总是与FHIT介导的细胞凋亡相关，并且FHIT基因转移可以消除体内的致瘤性（在H460和SK-Mes细胞中）或强烈抑制（在A549和SiHa细胞中）。我们的数据证明了Fhit蛋白在肺癌和宫颈癌细胞系中的抑癌特性和强促凋亡活性，并加强了其可能用作治疗工具的假设。
Loss of expression of the Fhit protein is often associated with the development of many human epithelial cancers, including lung and cervical carcinomas. Restoration of Fhit expression in cell lines derived from these tumors has however yielded conflicting results, prompting the need for careful evaluation of the oncosuppressive potential of FHIT. In the present study, we have investigated the effect of Fhit reintroduction in seven lung cancer and three cervical cancer cell lines. To achieve efficient gene transfer and high levels of transgene expression, we have used an adenoviral vector to transduce the FHIT gene. The induction of apoptosis was evaluated by using the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay and propidium iodide staining. Activation of caspases was detected by using Western blot analysis, and tumorigenic potential of transduced cells in the nude mouse was also assessed. Restoration of Fhit expression induced apoptosis in all Fhit-negative cell lines, with Calu-1, H460, and A549 being the most susceptible among the lung cancer cell lines and SiHa cells among cervical carcinomas. Activation of caspase-8 was always associated with Fhit-mediated apoptosis, and in vivo tumorigenicity was either abolished by FHIT gene transfer (in H460 and SK-Mes cells) or strongly suppressed (in A549 and SiHa cells). Our data demonstrate oncosuppressive properties and strong proapoptotic activity of the Fhit protein in lung and cervical cancer cell lines and strengthens the hypothesis of its possible use as a therapeutic tool.