Delivery of cytolethal distending toxin B induces cell cycle arrest and apoptosis in gingival squamous cell carcinoma in vitro.
Delivery of cytolethal distending toxin B induces cell cycle arrest and apoptosis in gingival squamous cell carcinoma in vitro.
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DOI：
10.1111/j.1600-0722.2004.00157.x
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发表时间：
2004-10
影响因子：
1.9
通讯作者：
Kozo Yamamoto;K. Tominaga;M. Sukedai;T. Okinaga;Kenjiro Iwanaga;T. Nishihara;J. Fukuda
中科院分区：
文献类型：
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作者：
Kozo Yamamoto;K. Tominaga;M. Sukedai;T. Okinaga;Kenjiro Iwanaga;T. Nishihara;J. Fukuda
伴放线放线杆菌（Actinobacillusactinomycetemcomitans）的细胞致死性膨胀毒素（cytolethalexpandingtoxin，Cdt）由CdtA、CdtB和CdtC三种蛋白组成，参与细胞周期阻滞和细胞凋亡。本研究以人牙龈鳞状细胞癌细胞系Ca 9 -22为靶细胞，建立了重组CdtB及CdtB表达质粒的局部给药系统。使用BioPORTER将CdtB导入Ca 9 -22细胞后，通过蛋白质印迹法检测到32-kDa蛋白质，发生G2期细胞周期阻滞和凋亡。此外，CdtB可上调Ca 9 -22细胞中磷酸化p53和细胞周期蛋白依赖性激酶抑制剂p21（CIP 1/WAF 1）的表达，提示这些细胞内分子可能参与了G2期细胞周期阻滞和细胞凋亡的诱导。当CdtB表达质粒通过脂质体转染或电穿孔转染到Ca 9 -22细胞中时，清晰地检测到CdtB（32 kDa）。此外，转染CdtB表达质粒后，观察到TdT介导的dUTP缺口末端标记阳性细胞。这些研究结果表明，交付的CdtB蛋白和cdtB基因的转染诱导细胞周期阻滞和凋亡的Ca 9 -22细胞在体外，我们的结论是，它可能是可能的诱导凋亡的人牙龈鳞状细胞癌通过电穿孔的cdtB基因。
The cytolethal distending toxin (Cdt) from Actinobacillus actinomycetemcomitans consists of three proteins, CdtA, CdtB, and CdtC, which are responsible for cell cycle arrest and apoptosis. In the present study, local delivery systems of recombinant CdtB and CdtB-expressing plasmid were established using Ca9-22, human gingival squamous cell carcinoma cell line. When CdtB was delivered to Ca9-22 cells using a BioPORTER, a 32-kDa protein was detected by Western blotting, and G2 cell cycle arrest and apoptosis occurred. In addition, the CdtB delivered upregulated the expression of phosphorylated p53 and the cyclin-dependent kinase inhibitor p21(CIP1/WAF1) in Ca9-22 cells, suggesting that these intracellular molecules might contribute to the induction of G2 cell cycle arrest and apoptosis. When the CdtB-expressing plasmid was transfected into Ca9-22 cells by lipofection or electroporation, CdtB (32 kDa) was clearly detected. Further, TdT-mediated dUTP nick end labeling positive cells were observed after transfection of the CdtB-expressing plasmid. These findings indicated that delivery of the CdtB protein and transfection of the cdtB gene induced cell cycle arrest and apoptosis in Ca9-22 cells in vitro, and we conclude that it may be possible to induce apoptosis in human gingival squamous cell carcinoma by electroporation of the cdtB gene.