天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2015年
9期
981-984,1093
,共5页
乳腺肿瘤%钙黏着糖蛋白类%肿瘤侵润%细胞运动%β-catenin%上皮间质转化%MDA-MB-231
乳腺腫瘤%鈣黏著糖蛋白類%腫瘤侵潤%細胞運動%β-catenin%上皮間質轉化%MDA-MB-231
유선종류%개점착당단백류%종류침윤%세포운동%β-catenin%상피간질전화%MDA-MB-231
breast neoplasms%cadherins%neoplasm invasiveness%cell movement%β-catenin%epithelial mesenchymal transition%MDA-MB-231
目的：探讨β-连环蛋白（β-catenin）对乳腺癌细胞系MDA-MB-231细胞上皮间质表型和侵袭迁移能力的影响。方法用β-catenin siRNA质粒转染乳腺癌MDA-MB-231细胞（对照组），免疫荧光染色观察β-catenin的表达情况。Western blot检测上皮标记蛋白E-cadherin、间质标记蛋白vimentin及上皮间质转化相关调控因子Twist1和Snail的表达。Transwell侵袭实验和伤口愈合实验比较对照组、空质粒组和β-catenin下调组细胞体外侵袭迁移能力。结果免疫荧光染色显示对照组和空质粒组细胞可见β-catenin在胞膜、胞浆和胞核表达，β-catenin下调组细胞胞膜β-catenin呈低表达，且未见胞浆、胞核表达。下调β-catenin的细胞E-cadherin表达增高，而vimentin、Twist1和Snail表达减少。侵袭迁移实验证明，下调组细胞穿膜细胞数和迁移距离显著少于MDA-MB-231和对照组(P＜0.05)。结论下调β-catenin阻碍Wnt/β-catenin通路的激活，降低乳腺癌MDA-MB-231细胞间质表型的表达而促进其上皮表型的表达，从而降低了细胞的体外侵袭、迁移能力。
目的：探討β-連環蛋白（β-catenin）對乳腺癌細胞繫MDA-MB-231細胞上皮間質錶型和侵襲遷移能力的影響。方法用β-catenin siRNA質粒轉染乳腺癌MDA-MB-231細胞（對照組），免疫熒光染色觀察β-catenin的錶達情況。Western blot檢測上皮標記蛋白E-cadherin、間質標記蛋白vimentin及上皮間質轉化相關調控因子Twist1和Snail的錶達。Transwell侵襲實驗和傷口愈閤實驗比較對照組、空質粒組和β-catenin下調組細胞體外侵襲遷移能力。結果免疫熒光染色顯示對照組和空質粒組細胞可見β-catenin在胞膜、胞漿和胞覈錶達，β-catenin下調組細胞胞膜β-catenin呈低錶達，且未見胞漿、胞覈錶達。下調β-catenin的細胞E-cadherin錶達增高，而vimentin、Twist1和Snail錶達減少。侵襲遷移實驗證明，下調組細胞穿膜細胞數和遷移距離顯著少于MDA-MB-231和對照組(P＜0.05)。結論下調β-catenin阻礙Wnt/β-catenin通路的激活，降低乳腺癌MDA-MB-231細胞間質錶型的錶達而促進其上皮錶型的錶達，從而降低瞭細胞的體外侵襲、遷移能力。
목적：탐토β-련배단백（β-catenin）대유선암세포계MDA-MB-231세포상피간질표형화침습천이능력적영향。방법용β-catenin siRNA질립전염유선암MDA-MB-231세포（대조조），면역형광염색관찰β-catenin적표체정황。Western blot검측상피표기단백E-cadherin、간질표기단백vimentin급상피간질전화상관조공인자Twist1화Snail적표체。Transwell침습실험화상구유합실험비교대조조、공질립조화β-catenin하조조세포체외침습천이능력。결과면역형광염색현시대조조화공질립조세포가견β-catenin재포막、포장화포핵표체，β-catenin하조조세포포막β-catenin정저표체，차미견포장、포핵표체。하조β-catenin적세포E-cadherin표체증고，이vimentin、Twist1화Snail표체감소。침습천이실험증명，하조조세포천막세포수화천이거리현저소우MDA-MB-231화대조조(P＜0.05)。결론하조β-catenin조애Wnt/β-catenin통로적격활，강저유선암MDA-MB-231세포간질표형적표체이촉진기상피표형적표체，종이강저료세포적체외침습、천이능력。
Objective To investigate the effect ofβ-catenin on epithelial mesenchymal phenotype and invasion migra?tion ability of breast cancer cell line MDA-MB-231. Methods Small interfering RNA (siRNA) targetingβ-catenin plas?mid was transfected into MDA-MB-231 cell line. Immunofluorescence staining was used to observe the expression of β-catenin. The expressions of epithelial cell marker E-cadherin and mesenchymal marker vimentin, epithelial mesenchymal transition correlation factor Twist1 and Snail were detected by Western blot assay. Invasion and migration ability was com?pared by transwell invasion and wound healing assay between control group, the MDA-MB-231 group andβ-catenin down-regulated group. Results Immunofluorescence staining showed thatβ-catenin was expressed in cell membrane, cytoplasm or nucleus in MDA-MB-231 group and control group. There was a decreased expression in β-catenin down-regulated group, and no expression in cytoplasm or nucleus. The expression of E-cadherin was increased, while vimentin, Twist 1 and Snail expression decreased inβ-catenin down-regulated cells. Transwell invasion and wound healing assay results proved that transmembrane cell number and migration distance were significantly lower in β-catenin down-regulated group than those of MDA-MB-231 group and control group (P<0.05). Conclusion The down-regulation ofβ-catenin inhibits Wnt/β-catenin activation that decreases the mesenchymal phenotype but increases epithelial phenotype of breast cancer cells MDA-MB-231, and which reduces the cell invasion and migration ability in vitro.
