HIV-TAT-fused FHIT protein functions as a potential pro-apoptotic molecule in hepatocellular carcinoma cells.
HIV-TAT-fused FHIT protein functions as a potential pro-apoptotic molecule in hepatocellular carcinoma cells.
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HIV-TAT 融合的 FHIT 蛋白在肝细胞癌细胞中充当潜在的促凋亡分子。
DOI：
10.1042/bsr20110033
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发表时间：
2012-06
影响因子：
4
通讯作者：
等
中科院分区：
文献类型：
--
作者：
余桂戎;秦炜炜;李纪鹏;等
越来越多的证据表明，FHIT（脆性组氨酸三联体）是一个真正的肿瘤抑制基因在很大一部分人类肿瘤，包括肝细胞癌。已经开发了基于病毒的递送系统来将FHIT基因转移到多种类型的癌细胞中以抑制生长甚至诱导凋亡。然而，基于蛋白质的FHIT替代策略尚未在癌细胞中进行。在这里，我们使用HIV-TAT（转录反式激活因子）衍生肽转移纯化的FHIT蛋白到肝癌（肝细胞癌）细胞，并确定这种融合蛋白在诱导凋亡的生物学效应。采用亲和层析法从大肠杆菌BL 21中纯化达特肽融合人FHIT蛋白（TAT-FHIT）。通过免疫荧光染色和Western blot分析，以鉴定TAT-FHIT在HCC细胞中的表达和内化，并与纯化的FHIT蛋白进行比较。我们的研究表明，TAT-FHIT蛋白在37°C孵育后1 h内可转位到癌细胞中。MTT法、Annexin-V染色和Western blotting检测结果表明TAT-FHIT能显著抑制肝癌细胞的生长并诱导其凋亡。此外，一项机制研究表明，外源性和内源性凋亡途径都参与了TAT-FHIT介导的细胞凋亡，并且这种作用可以被线粒体保护剂TAT-BH 4部分减弱，表明TAT-BH 4在癌细胞中TAT-FHIT介导的促凋亡作用中起关键作用。总之，我们的研究表明，TAT-FHIT是一个潜在的促肝癌细胞凋亡分子，并加强其治疗应用于肝癌的假设。
Accumulating evidence has demonstrated that FHIT (fragile histidine triad) is a bona fide tumour suppressor gene in a large fraction of human tumours, including hepatocellular cancer. A virus-based delivery system has been developed to transfer the FHIT gene into many types of cancer cells to inhibit growth or even induce apoptosis. However, a protein-based replacement strategy for FHIT has not been performed in cancer cells. Here, we used HIV-TAT (transactivator of transcription)-derived peptide to transfer the purified FHIT protein into HCC (hepatocellular carcinoma) cells and determine the biological effect of this fusion protein in inducing apoptosis. Affinity chromatography was used to purify TAT peptide-fused human FHIT (TAT-FHIT) protein from BL21 Escherichia coli. Immunofluorescence staining and Western blot analysis were performed to identify the expression and internalization of TAT-FHIT in HCC cells compared with the purified FHIT protein. Our study showed that TAT-FHIT protein can translocate into cancer cells in 1 h after incubation at 37°C. Furthermore, the results of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, Annexin-V staining and Western blotting demonstrated that TAT-FHIT can robustly inhibit growth and induce apoptosis of HCC cells in vitro. In addition, a mechanistic study showed that both exogenous and intrinsic apoptotic pathways were involved in TAT-FHIT-mediated apoptosis and this effect could be attenuated partially by a mitochondrial protector TAT-BH4, indicating that mitochondrion plays a critical role in TAT-FHIT-mediated pro-apoptotic effect in cancer cells. Taken together, our study suggests that TAT-FHIT is a potential pro-apoptotic molecule in HCC cells and strengthen the hypothesis of its therapeutic application against HCC.
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DOI：
10.1073/pnas.191345898
发表时间：
2001-08-28
影响因子：
11.1
作者：
Zanesi, N;Fidanza, V;Huebner, K
通讯作者：
Huebner, K
影响因子：
4.5
作者：
Lachenmayer, Anja;Alsinet, Clara;Chang, Charissa Y.;Llovet, Josep M.
通讯作者：
Llovet, Josep M.
影响因子：
1.6
作者：
T. Pawełczyk;R. Kowara;F. Golebiowski;A. Matecki
通讯作者：
T. Pawełczyk;R. Kowara;F. Golebiowski;A. Matecki
影响因子：
2.7
作者：
J. Zawacka-Pankau;A. Podhajska
通讯作者：
J. Zawacka-Pankau;A. Podhajska
FHIT基因敲除小鼠中自发性肿瘤和肠道病变的发展。
影响因子：
8.8
作者：
Fujishita, T;Doi, Y;Sonoshita, M;Hiai, H;Oshima, M;Huebner, K;Croce, CM;Taketo, MM
通讯作者：
Taketo, MM