c—Met抑制剂SU11274对基底样乳腺癌MDA-MB-231细胞的影响  

Effects of c-Met inhibitor SUl1274 on basal-like breast cancer cells MDA-MB-231

作　　者：冯炜红[1] 张斌[1] 李媛媛[1] 赵洪猛[1] 张月[1] 陈祖锦[1] 刘博文[1] 曹旭晨[1] 

机构地区：[1]天津医科大学附属肿瘤医院乳腺一科天津市肿瘤防治重点实验室乳腺癌防治教育部重点实验室,300060

出　　处：《中华普通外科杂志》2012年第3期234-237,共4页Chinese Journal of General Surgery

基　　金：国家自然科学基金资助项目（81001186）;天津市自然科学基金资助项目（10JCYBJC14100）

摘　　要：目的研究c-Met抑制剂SUll274对c—Met阳性基底样乳腺癌细胞系MDA—MB-231凋亡和运动的影响。方法荧光染料Hoechst33342、MitroTrackerRed和Yo-pro-1染色，观察SU11274诱导细胞凋亡的形态学变化；流式细胞术检测不同处理组的细胞早期凋亡率；Westernblot检测凋亡相关蛋白的表达变化和c．Met及Akt磷酸化水平的改变；划痕试验及趋化试验观察SU11274对细胞迁移、趋化能力的影响。结果SUll274（10ixmol／L）处理MDA—MB-231细胞48h后，与对照组比较，荧光染色可见处理组细胞核绿染，胞核碎裂；各加药组MDA—MB-231的早期凋亡率分别为（7．3±0．9）％、（14．1±0．6）％、（35．5±4．4）％、（48．2±5．3）％，与对照组相比明显升高（P〈0．05）。同时，抗凋亡蛋白Bcl．XL表达减少，凋亡相关蛋白Caspase-3和PARP蛋白剪切增加，量效关系明显。SUll274可显著延长划痕愈合时间以及减少趋化穿膜细胞个数（P〈0．05），并有效抑制c-Met及Akt的磷酸化水平，呈剂量依赖性关系。结论SU11274可通过抑制c—Met／P13K／Akt的磷酸化水平而诱导c-Met阳性基底样乳腺癌细胞系MDA-MB-231凋亡，并抑制其运动。Objective To investigate the effects of a new c-Met inhibitor SUl1274 on apoptosis and motility of c-Met-positive basal-like breast cancer cells MDA-MB-231. Methods The concentrations of SUl1274 were set to 0, 0. 1, 1, 10 and 20 μmol/L. Morphological change of apoptotic cells was analyzed by Hoechst33342, MitroTrackerRed and Yo-pro-1 staining. The apoptotic rate of MDA-MB-231 ceils were determined by Annexin V/PI double-staining. The expression of apoptosis related proteins （Bcl-XL, Caspase-3 and PARP） and phosphorylation levels of c-Met and Akt were analyzed by Western blot. The capability of motility were measured by wound-healing assay and chemotaxis assay. Results After treatment by SUl1274（10 μmol/L） for 48 h, shrinking apoptotic cells of MDA-MB-231 was observed by flurescent microscope and nuclear fragmentation was seen. Annexin V/PI double-staining showed SU11274 induced apoptosis of MDA-MB-231 cells （P 〈 0. 05 ）, and the apoptotic rates were （7.3 ± 0.9） %, （ 14. 1 ± 0. 6） %, （35. 5 ± 4. 4） % and （48. 2 ± 5. 3） %, respectively. SU11274 downregulated the expression of Bcl- XL and promoted the dissection of Caspase-3 and PARP in a dose dependent relationship. SU11274 prolongs the wound-healing time, decreases the migration cell count （ P 〈 0. 05 ） and effectively inhibits the phosphorylation of c-Met and its downstream key proteins Akt in a dose-dependent manner. Conclusions C-Met inhibitor SU11274 induces apoptosis and inhibits the motility of c-Met-positive basal-like breast cancer cell line MDA-MB-231, probably through inhibiting phosphorylation of c-Met/PI3K/Akt.

关 键 词：乳腺肿瘤 癌 基底细胞 原癌基因蛋白质C-MET 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]