Silencing of galectin-3 changes the gene expression and augments the sensitivity of gastric cancer cells to chemotherapeutic agents
Silencing of galectin-3 changes the gene expression and augments the sensitivity of gastric cancer cells to chemotherapeutic agents
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DOI：
10.1111/j.1349-7006.2009.01364.x
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发表时间：
2010-01-01
期刊：
影响因子：
5.7
通讯作者：
Chun, Kyung-Hee
中科院分区：
文献类型：
--
作者：
Cheong, Teak-Chin;Shin, Ji-Young;Chun, Kyung-Hee
Galectin-3 已知可调节细胞增殖和凋亡，并且在人类癌症中高表达，但其在胃癌中的功能仍存在争议。在这里，我们通过使用合成的双链 siRNA 沉默半乳糖凝集素 3 在胃癌细胞中的作用。半乳糖凝集素3沉默后，细胞数量减少，细胞形状发生变化。 Galectin-3 siRNA 处理也诱导 G(1) 停滞。 DNA 微阵列分析用于评估 galectin-3 沉默后基因表达的变化。我们发现半乳糖凝集素 3 的沉默会导致基因表达的变化。 RT-PCR 和实时 PCR 用于验证微阵列研究中发现的变化。蛋白质印迹分析证实了感兴趣的蛋白质表达的变化：细胞周期蛋白 D1、生存素、XIAP、XAF、PUMA 和 GADD45 α。一般来说，它倾向于增加几种促凋亡基因的表达，并减少细胞周期进展基因的表达。我们还证实这些基因表达的变化是由galectin-3过度表达引起的。最后，我们证明，沉默galectin-3可通过进一步降低抗凋亡和/或细胞存活分子（如生存素、细胞周期蛋白D1和XIAP）的表达并增加促凋亡XAF-1的表达来增强化疗药物对细胞凋亡的诱导。我们的结论是，galectin-3通过调节多个相关基因的表达参与癌症进展和恶性肿瘤，抑制galectin-3可能是改善胃癌化疗的一种方法。 （癌症科学 2009）。
Galectin-3 is known to modulate cell proliferation and apoptosis and is highly expressed in human cancers, but its function in gastric cancer is still controversial. Here, we examined the role of galectin-3 in gastric cancer cells by silencing it with synthetic double-stranded siRNA. After silencing of galectin-3, cell numbers decreased and cell shape changed. Galectin-3 siRNA treatment also induced G(1) arrest. DNA microarray analysis was used to assess changes in gene expression following galectin-3 silencing. We found that silencing of galectin-3 caused changes in gene expression. RT-PCR and real-time PCR were utilized for validation of the changes found in microarray studies. Western blot analysis confirmed changes in the expression of proteins of interest: cyclin D1, survivin, XIAP, XAF, PUMA, and GADD45 alpha. Generally, it tended to increase the expression of several pro-apoptotic genes, and to decrease the expression of cell cycle progressive genes. We also confirmed that changes in the expression of these genes were caused by galectin-3 overexpression. Finally, we demonstrated that silencing of galectin-3 enhanced apoptosis induction with chemotherapeutic agents by further reducing the expression of anti-apoptotic and/or cell survival molecules such as survivin, cyclin D1, and XIAP, and increasing the expression of pro-apoptotic XAF-1. We conclude that galectin-3 is involved in cancer progression and malignancy by modulating the expression of several relevant genes, and inhibition of galectin-3 may be an approach to improve chemotherapy of gastric cancers. (Cancer Sci 2009).