促肝再生磷酸酶-3拮抗剂原钒酸钠对大肠癌细胞增殖凋亡影响  

The effects of phosphatase of regeneration liver-3 inhibitor sodium orthovanadate on proliferation and apoptosis of Colo-320 cells

作　　者：赵高平[1] 周总光[2] 郑雪莲[2] 王存[2] 

机构地区：[1]四川省医学科学院四川省人民医院胃肠外科,成都610072 [2]四川大学华西医院消化外科研究室

出　　处：《中华实验外科杂志》2008年第11期1438-1440,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨原钒酸钠（SoV）通过对促肝再生磷酸酶-3（PRL-3）抑制，观察对大肠癌细胞株Colo-320增殖凋亡影响。方法应用Western blot方法检测PRL-3在7株大肠癌细胞中表达情况，筛选出表达最强的一株做下一步抑制实验。选择PRL-3拮抗剂SoV，通过噻唑蓝（MTT）实验观察其在0．1～0．5μmol／L浓度范围对癌细胞增殖影响；通过流式细胞术计算增殖指数（PI），观察其对癌细胞周期和凋亡影响。结果Western blot方法筛选出表达PRL-3最强结肠癌细胞株Colo-320，MTT实验显示SoV能显著抑制Colo-320细胞生长能力，呈量-效（0．1—0．5μmol／L）、时-效（0～120h）相关性。0．5μmol／L SoV作用48h后，S期细胞比例为（17．5±0．6）％，与对照组S期所占比例（28．0±1．1）％比较明显下降（P〈0．01），细胞分裂阻滞在S期，PI值亦明显降低（P〈0．01），没有发现细胞凋亡。结论SoV能显著抑制Colo-320细胞增殖，其机制与抑制PRL-3酶活性以及参与细胞周期调控有关，而与细胞凋亡过程无关。Objective To elucidate the effects of sodium orthovanadate （SoV）, an inhibitor of PRL-3 phosphatase ,on proliferation and apoptosis of Colo-320 cells through blocking the catalytical activity of PRL-3 phosphatase. Methods The expression levels of PRL-3 in 7 colorectal cancer cell lines were examined by Western blot in order to screen out which cell line expressed the highest level of PRL-3 protein. Then this cell line was employed in the next experiments of drug intervention. MTT assay was used to analyze the effects of SoV on proliferation of cancer cells after the cells were exposed to SoV at various concentrations of 0.1,0. 2 ,0. 3 ,0. 4,0. 5 μmol/L, respectively, for 0 to 120 h. The optimal concentration and time were chosen to test the effects of SoV on the cancer cell cycle and apoptosis by flow cytometry. Results The expression of PRL-3 was clearly detected in 5 of 7 （71.4%） colorectal cancer cell lines by Western blot. The colonic cancer cell line Colo-320 was screened as the target ceil for its highest expression level of PRL-3. MTT assay showed that SoV inhibited the proliferation of cancer cells in a dose- and time-dependent manner. After exposure to SoV at a concentration of 0.5 μmol/L for 48 h, the percentage of ceils, （ 17.5 ± 0.6 ） % ,in S-phase was significantly decreased as compared to that of （28.0 ± 1.1 ） % in control ceils （P 〈 0.01 ） , indicating that the cell division was blocked in S-phase. No obvious signs of apoptosis were observed.. Conclusion SoV shows good inhibition efficacy to colonic cancer cell growth, which may be yielded through restraining catalytical activity of PRL-3 phosphatase and then blocking the cell cycle in S-phase.

关 键 词：促肝再生磷酸酶-3 大肠癌 脱噬作用 

分 类 号：R735.34[医药卫生—肿瘤]