中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
2期
297-299
,共3页
乳腺癌%转化生长因子-β1%脂多糖%肿瘤转移%上皮-间充质转化
乳腺癌%轉化生長因子-β1%脂多糖%腫瘤轉移%上皮-間充質轉化
유선암%전화생장인자-β1%지다당%종류전이%상피-간충질전화
Breast cancer%Transforming growth factor-β1%Lipopolysaccharide%Tumor metastasis%Epithelial-mesenchymal transition
目的 观察脂多糖(LPS)协同转化生长因子-β1 (TGF-β1)作用于乳腺癌MCF-7细胞的生物学效应,并探讨其分子生物学机制.方法 倒置显微镜下观察细胞形态;罗丹明-鬼笔环肽标记细胞骨架;实时定量聚合酶链反应(Real-time PCR)检测E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、转录因子Snail、Twist以及基质金属蛋白酶-9(MMP-9)的基因表达水平;Transwell小室检测迁移、侵袭能力;明胶酶谱检测活性蛋白MMP-9的表达水平;Western blot检测核因子(NF)-κB、细胞外信号调节激酶(ERK)和Smad信号通路.结果 LPS和TGF-β1联合组(联合刺激组)的细胞呈纺锤状,细胞骨架明显改变.联合刺激组E-cadherin显著下调,Vimentin、Snail-2、Twist、MMP-9 mRNA表达水平显著上调;对照组、LPS组、TGF-β1组、联合刺激组每个视野迁移细胞数分别为(6.8±4.1)、(10.2±4.3)、(39.5士3.8)、(69.7±4.4)个,每个视野侵袭细胞数分别为(4.4±1.1)、(6.8±2.4)、(32.1±2.3)、(54.9±4.5)个.联合刺激组活性MMP-9表达水平最高;联合刺激组磷酸化ERK和磷酸化Smad-2的水平要高于TGF-β1组.结论 LPS能够增强TCF-β1诱导乳腺癌MCF-7细胞发生上皮-间充质转化,并且两者联合作用能够促进MCF-7细胞发生侵袭迁移.
目的 觀察脂多糖(LPS)協同轉化生長因子-β1 (TGF-β1)作用于乳腺癌MCF-7細胞的生物學效應,併探討其分子生物學機製.方法 倒置顯微鏡下觀察細胞形態;囉丹明-鬼筆環肽標記細胞骨架;實時定量聚閤酶鏈反應(Real-time PCR)檢測E-鈣黏蛋白(E-cadherin)、波形蛋白(Vimentin)、轉錄因子Snail、Twist以及基質金屬蛋白酶-9(MMP-9)的基因錶達水平;Transwell小室檢測遷移、侵襲能力;明膠酶譜檢測活性蛋白MMP-9的錶達水平;Western blot檢測覈因子(NF)-κB、細胞外信號調節激酶(ERK)和Smad信號通路.結果 LPS和TGF-β1聯閤組(聯閤刺激組)的細胞呈紡錘狀,細胞骨架明顯改變.聯閤刺激組E-cadherin顯著下調,Vimentin、Snail-2、Twist、MMP-9 mRNA錶達水平顯著上調;對照組、LPS組、TGF-β1組、聯閤刺激組每箇視野遷移細胞數分彆為(6.8±4.1)、(10.2±4.3)、(39.5士3.8)、(69.7±4.4)箇,每箇視野侵襲細胞數分彆為(4.4±1.1)、(6.8±2.4)、(32.1±2.3)、(54.9±4.5)箇.聯閤刺激組活性MMP-9錶達水平最高;聯閤刺激組燐痠化ERK和燐痠化Smad-2的水平要高于TGF-β1組.結論 LPS能夠增彊TCF-β1誘導乳腺癌MCF-7細胞髮生上皮-間充質轉化,併且兩者聯閤作用能夠促進MCF-7細胞髮生侵襲遷移.
목적 관찰지다당(LPS)협동전화생장인자-β1 (TGF-β1)작용우유선암MCF-7세포적생물학효응,병탐토기분자생물학궤제.방법 도치현미경하관찰세포형태;라단명-귀필배태표기세포골가;실시정량취합매련반응(Real-time PCR)검측E-개점단백(E-cadherin)、파형단백(Vimentin)、전록인자Snail、Twist이급기질금속단백매-9(MMP-9)적기인표체수평;Transwell소실검측천이、침습능력;명효매보검측활성단백MMP-9적표체수평;Western blot검측핵인자(NF)-κB、세포외신호조절격매(ERK)화Smad신호통로.결과 LPS화TGF-β1연합조(연합자격조)적세포정방추상,세포골가명현개변.연합자격조E-cadherin현저하조,Vimentin、Snail-2、Twist、MMP-9 mRNA표체수평현저상조;대조조、LPS조、TGF-β1조、연합자격조매개시야천이세포수분별위(6.8±4.1)、(10.2±4.3)、(39.5사3.8)、(69.7±4.4)개,매개시야침습세포수분별위(4.4±1.1)、(6.8±2.4)、(32.1±2.3)、(54.9±4.5)개.연합자격조활성MMP-9표체수평최고;연합자격조린산화ERK화린산화Smad-2적수평요고우TGF-β1조.결론 LPS능구증강TCF-β1유도유선암MCF-7세포발생상피-간충질전화,병차량자연합작용능구촉진MCF-7세포발생침습천이.
Objective To investigate into the biological effect of lipopolysaccharide (LPS) combined with transforming growth factor-β1 (TGF-β1) on MCF-7 breast cancer cells and discuss the molecular mechanism underneath the phenomena.Methods The morphology of MCF-7 cells was observed under the inverted microscope.Rhodamine-phalloidine was used for staining the actin cytoskeleton.Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect genetic expression of E-cadherin,Vimentin,Snail-2,Twist and matrix metalloproteinase-9 (MMP-9).Transwell was used to examine cell migration and invasion.Zymography experiment was used to test active protein MMP-9.Western blotting was used to detect signal pathways of nuclear factor-κB (NF-κB),extracellular signal-regulated kinase (ERK) and Smad.Results Almost all the cells in the group of LPS combined with TGF-β1 (L + T group) acquired fibroblastoid properties along with obvious cytoskeleton changes.L + T group had E-cadherin down-regulated obviously while Snail-2,Twist and MMP-9 up-regulated sharply.The number of migrated cells in control group,LPS group,TGF-β1 group,L + T group were 6.8 ±4.1,10.2 ±4.3,39.5 ±3.8,69.7 ±4.4 respectively.The number of invaded cells in control group,LPS group,TGF-β1 group,L + T group were 4.4 ± 1.1,6.8 ± 2.4,32.1 ± 2.3,57.9 ± 4.5 respectively.The active protein of MMP-9 was the most in L + T group.L + T group has much higher phosphorylated ERK and phosphorylated Smad-2 than TGF-β1 group's.Conclusion LPS cooperates with TGF-β1 to promote the epithelial to mesenchymal transition of MCF-7 breast cancer cells,and stimulation of LPS and TGF-β1 on MCF-7 make those cells have stronger capabilities of migration and invasion.
