Abstract:

Objective To investigate role of TGF?beta in promoting the colorectal cancer LoVo cells invasion induced by phosphatase of regenerating liver-3（PRL?3）. Methods ELISA assay was to used to detect the level of TGF-beta in LoVo-P and LoVo-C. Invasion assays were applied to determine the effect of TGF-beta on the ability of PRL-3. Western blot was used to detect the proteins of p-AKT and AKT. Results Elisa assay displayed that the protein level of TGF?beta of LoVo?P was higher than LoVo-C （114 ± 11 pg/ml vs. 56 ± 8 pg/ml, P<0.01）. When added different dosages of neutralizing antibody of TGF-beta （1, 10, 00 ng/ml） in culture medium of LoVo-P, the invasion were decreased significantly（96.1 ± 8.2, 67.3 ± 9.4 and 48.6 ± 6.4, respectively）and all Pvaluesless than 0.05）. Western blot reminded the proteins of p?AKT in LoVo?P were higher than LoVo?C, and the expression of p?AKT in LoVo?P was decreased by 2.5?fold after treating LoVo?P with the PI3K inhibitor LY294002（10 mg/mL） （P<0.05）. Conclusion PRL?3 could up?regulate the expression of TGF?beta in colorectal cancer LoVo cells and promote the invasion of LoVo cells, in which PI3K/AKT signaling pathway may be involved.

Key words: PRL?3, invasion, colon cancer, TGF?beta

摘要：

目的 探讨转化生长因子-beta（TGF?beta）通路在肝再生磷酸酶?3（PRL?3）促进结肠癌细胞侵袭和转移中的作用机制。方法 酶联免疫吸附试验ELISA检测实验组与对照组结肠癌细胞TGF?beta水平表达差异，Transwell侵袭小室法检测LoVo细胞侵袭能力变化，Westernblot检测相关通路蛋白水平变化。结果 ELISA实验检测结肠癌细胞转染PRL?3后，稳定表达PRL?3的LoVo细胞（LoVo?P）与对照组（LoVo?C）相比，TGF?beta表达明显升高（114±11pg/mL比56±8pg/mL，P<0.01）。Transwell侵袭实验提示不同浓度的TGF?beta中和抗体（1、10、100ng/mL）作用LoVo?P后其侵袭能力逐渐降低，对照组穿膜细胞数为142.7±11.3个，实验组分别为96.1±8.2、67.3±9.4、48.6±6.4个（P<0.05）。蛋白印记实验提示PI3K/AKT信号通路参与了其诱导侵袭的过程，使用PI3K抑制剂LY294002（10mg/mL）后，LoVo?P的AKT磷酸化蛋白的表达水平降低2.5倍（P<0.05）。结论 PRL?3能诱导结肠癌LoVo细胞分泌TGF?beta，激活PI3K/AKT信号通路进而促进结肠癌LoVo细胞侵袭。

关键词: 结肠癌, 转化生长因子-beta, 肝再生磷酸酶-3, 侵袭