中华普通外科杂志
中華普通外科雜誌
중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2012年
3期
234-237
,共4页
冯炜红%张斌%李媛媛%赵洪猛%张月%陈祖锦%刘博文%曹旭晨
馮煒紅%張斌%李媛媛%趙洪猛%張月%陳祖錦%劉博文%曹旭晨
풍위홍%장빈%리원원%조홍맹%장월%진조금%류박문%조욱신
乳腺肿瘤%癌,基底细胞%原癌基因蛋白质c-met%凋亡
乳腺腫瘤%癌,基底細胞%原癌基因蛋白質c-met%凋亡
유선종류%암,기저세포%원암기인단백질c-met%조망
Breast neoplasms%Carcinoma,basal cell%Proto-oncogene proteins c-met%Apoptosis
目的 研究c-Met抑制剂SU11274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SU 11274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;Western blot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SU 11274对细胞迁移、趋化能力的影响.结果 SU11274(10 μmol/L)处理MDA-MB-231细胞48 h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.SU11274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/PI3 K/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.
目的 研究c-Met抑製劑SU11274對c-Met暘性基底樣乳腺癌細胞繫MDA-MB-231凋亡和運動的影響.方法 熒光染料Hoechst33342、MitroTrackerRed和Yo-pro-1染色,觀察SU 11274誘導細胞凋亡的形態學變化;流式細胞術檢測不同處理組的細胞早期凋亡率;Western blot檢測凋亡相關蛋白的錶達變化和c-Met及Akt燐痠化水平的改變;劃痕試驗及趨化試驗觀察SU 11274對細胞遷移、趨化能力的影響.結果 SU11274(10 μmol/L)處理MDA-MB-231細胞48 h後,與對照組比較,熒光染色可見處理組細胞覈綠染,胞覈碎裂;各加藥組MDA-MB-231的早期凋亡率分彆為(7.3±0.9)％、(14.1±0.6)％、(35.5±4.4)％、(48.2±5.3)％,與對照組相比明顯升高(P＜0.05).同時,抗凋亡蛋白Bcl-XL錶達減少,凋亡相關蛋白Caspase-3和PARP蛋白剪切增加,量效關繫明顯.SU11274可顯著延長劃痕愈閤時間以及減少趨化穿膜細胞箇數(P＜0.05),併有效抑製c-Met及Akt的燐痠化水平,呈劑量依賴性關繫. 結論 SU11274可通過抑製c-Met/PI3 K/Akt的燐痠化水平而誘導c -Met暘性基底樣乳腺癌細胞繫MDA-MB-231凋亡,併抑製其運動.
목적 연구c-Met억제제SU11274대c-Met양성기저양유선암세포계MDA-MB-231조망화운동적영향.방법 형광염료Hoechst33342、MitroTrackerRed화Yo-pro-1염색,관찰SU 11274유도세포조망적형태학변화;류식세포술검측불동처리조적세포조기조망솔;Western blot검측조망상관단백적표체변화화c-Met급Akt린산화수평적개변;화흔시험급추화시험관찰SU 11274대세포천이、추화능력적영향.결과 SU11274(10 μmol/L)처리MDA-MB-231세포48 h후,여대조조비교,형광염색가견처리조세포핵록염,포핵쇄렬;각가약조MDA-MB-231적조기조망솔분별위(7.3±0.9)％、(14.1±0.6)％、(35.5±4.4)％、(48.2±5.3)％,여대조조상비명현승고(P＜0.05).동시,항조망단백Bcl-XL표체감소,조망상관단백Caspase-3화PARP단백전절증가,량효관계명현.SU11274가현저연장화흔유합시간이급감소추화천막세포개수(P＜0.05),병유효억제c-Met급Akt적린산화수평,정제량의뢰성관계. 결론 SU11274가통과억제c-Met/PI3 K/Akt적린산화수평이유도c -Met양성기저양유선암세포계MDA-MB-231조망,병억제기운동.
Objective To investigate the effects of a new c-Met inhibitor SU11274 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 cells 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 SU11274( 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 SU11274induced 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 basallike breast cancer cell line MDA-MB-231,probably through inhibiting phosphorylation of c-Met/PI3K/Akt.
