Growth inhibitory effects and molecular mechanisms of crotoxin treatment in esophageal Eca-109 cells and transplanted tumors in nude mice
Growth inhibitory effects and molecular mechanisms of crotoxin treatment in esophageal Eca-109 cells and transplanted tumors in nude mice
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响尾蛇毒素处理食管Eca-109细胞及裸鼠移植瘤的生长抑制作用及分子机制
DOI：
10.1038/aps.2012.156
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发表时间：
2012-12
影响因子：
8.2
通讯作者：
Xie, Yan
中科院分区：
文献类型：
--
作者：
He, Jing-kang;Wu, Xiang-sheng;Wang, Yan;Han, Rong;Qin, Zheng-hong;Xie, Yan
目的：研究硬毛响尾蛇神经毒素（crotoxin）对体外培养的人食管癌Eca-109细胞及裸鼠食管癌Eca-109移植瘤的抑制作用。Hoechst 33342染色观察细胞核形态变化，流式细胞仪检测细胞凋亡和细胞周期分布。RT-PCR检测Bcl-2、p15、caspase-3、p17基因表达水平。将Eca-109细胞接种于雌性Balb/c裸鼠体内建立肿瘤移植模型。将Crotoxin（25、50和100 mg/kg）皮下注射到移植的肿瘤中，每2天一次，共注射10次。测量肿瘤大小和重量。结果：Crotoxin（25、50和100 μg/mL）对Eca-109细胞的生长抑制率分别为22.9%、35.8%和57.2%，呈剂量依赖性; Hoechst 33342染色显示crotoxin处理后细胞核染色质固缩的凋亡细胞。crotoxin可诱导Eca-109细胞凋亡和G1期阻滞，并显著上调p15和caspase-3 p17基因的表达，下调Bcl-2基因的表达。此外，crotoxin抑制Eca-109裸鼠肿瘤的生长在剂量依赖性的方式。Western blotting结果显示，crotoxin可上调肿瘤组织中p15和caspase-3 p17蛋白的表达，下调Bcl-2蛋白的表达。结论：Crotoxin可通过诱导Eca-109细胞凋亡和阻滞G1期来抑制Eca-109细胞生长。crotoxin局部给药抑制裸鼠皮下移植Eca-109细胞的生长，可能通过增加p15和caspase-3 p17蛋白表达，降低Bcl-2蛋白表达。
Aim:To investigate the antitumor actions of the Crotalus durissus neurotoxin (crotoxin) on human esophageal carcinoma (Eca-109) cells in vitro and transplanted esophageal Eca-109 tumors in nude mice.Methods:The growth-inhibitory effect was analyzed in Eca-109 cells using MTT assay. Cell morphology changes in nuclei were observed using Hoechst 33342 staining, while apoptosis and cell cycle distribution were examined by flow cytometry. RT-PCR was used to measure the Bcl-2, p15, and caspase-3 p17 gene expression levels. A tumor transplantation model was established by inoculation of Eca-109 cells were into female Balb/c nude mice. Crotoxin (25, 50, and 100 mg/kg) was subcutaneously injected into the transplanted tumors every 2 d for a total of 10 injections. Tumor size and weight were measured. Bcl-2, p15, and caspase-3 p17 protein expression in transplanted tumors was analyzed using Western blotting.Results:Crotoxin (25, 50, and 100 μg/mL) inhibited the growth of Eca-109 cells in a dose-dependent manner with inhibition rates of 22.9%, 35.8%, and 57.2%, respectively. Hoechst 33342 staining revealed apoptotic cells with pyknotic nuclear chromatin after crotoxin treatment. In Eca-109 cells, crotoxin induced apoptosis and G1 block, significantly upregulated the expression of p15 and caspase-3 p17 genes and downregulated the expression of Bcl-2 gene. Furthermore, crotoxin inhibited the growth of Eca-109 tumors in nude mice in a dose-dependent manner. Western blotting showed that crotoxin increased p15 and caspase-3 p17 protein levels and reduced Bcl-2 protein level in tumor specimens.Conclusion:Crotoxin inhibits the growth of Eca-109 cells in vitro via apoptosis induction and G1 block. Local administration of crotoxin inhibits the growth of subcutaneously transplanted Eca-109 cells in nude mice, possibly via increasing p15 and caspase-3 p17 protein expression and reducing Bcl-2 protein expression.
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影响因子：
3.6
作者：
Baskar R;Lee KA;Yeo R;Yeoh KW
通讯作者：
Yeoh KW
DOI：
10.1016/j.toxicon.2006.01.010
发表时间：
2006-04
期刊：
Toxicon : official journal of the International Society on Toxinology
影响因子：
--
作者：
Ci-hui Yan;Zhong-Qin Liang;Z. Gu;Ya-ping Yang;P. Reid;Zheng-Hong Qin
通讯作者：
Ci-hui Yan;Zhong-Qin Liang;Z. Gu;Ya-ping Yang;P. Reid;Zheng-Hong Qin
影响因子：
2.9
作者：
Andres Pereanez, Jaime;Dario Gomez, Ivan;Camilo Patino, Arley
通讯作者：
Camilo Patino, Arley
影响因子：
0.2
作者：
A. Mayora;F. Arvelo
通讯作者：
A. Mayora;F. Arvelo
影响因子：
8.2
作者：
Ci-hui Yan;Ya-ping Yang;Zheng-Hong Qin;Z. Gu;P. Reid;Zhong-Qin Liang
通讯作者：
Ci-hui Yan;Ya-ping Yang;Zheng-Hong Qin;Z. Gu;P. Reid;Zhong-Qin Liang