摘要
目的探讨TGF-β1诱导大肠癌HT29细胞发生上皮细胞间质转化(EMT)后对肿瘤细胞侵袭转移能力的影响。方法 TGF-β1长期诱导HT29细胞,RT-PCR检测细胞E-钙黏素和波形蛋白mRNA的表达;HE和考马斯亮蓝染色后分别观察细胞形态和骨架;对HT29、经TGF-β1诱导发生EMT的HT29/TGF-β1和两者混合培养的细胞分别进行划痕实验;绿色荧光蛋白标记质粒转染HT29/TGF-β1细胞,红色的转染HT29细胞,两者混合培养进行划痕实验。结果 TGF-β1连续刺激10 d后HT29的Ecadherin表达显著下降,Vimentin表达显著上调(P<0.05);连续刺激40 d后,细胞变梭,细胞间连接不紧密。细胞蓝色丝网状结构的骨架蛋白明显增多,相邻细胞间骨架连接不如刺激前紧密;细胞划痕实验显示刺激后的HT29细胞迁移速度最快,HT29细胞最慢;转染后见红色和绿色的细胞在迁移生长的过程中分布均匀。结论 TGF-β1可诱导大肠癌细胞发生EMT;大肠癌中发生EMT的细胞可以加速非EMT细胞的侵袭转移速度。
Objective To explore the biological behaviors of EMT cells and non-EMT cells in colorectal cancer metastasis. Methods TGF-β1was used to stimulate colon cancer cells HT29. The expression of Ecadherin and Vimentin mRNA of cells was detected by fluorescence quantitative PCR. The morphology and Cytoskeleton of HT29 and HT29/TGF-β1 cells were observed by HE and Coomassie blue staining respectively, the green and red fluorescent protein plasmid were transfected into EMT and non-EMT cells respectively, then mixed culturing two kind cells, doing scratch test, observing the movement the of cells. Results After continuous stimulated by TGF-β1 for 10 d, the expression of Ecadherin mRNA decreased with significant statistical difference, the expression of Vimentin mRNA increased significanty statistical（P 〈0. 05）. After continuous stimulation by TGF-β1 for 40 d, HT29 cells showed the morphology of with spindle declined cell-cell adhesion. HT29 cells have more surface projections, and less cytoskeletal structures and spot-like actin bodies than HT29/TGF-β1 cells. Scratch test shows that the migration rate of the HT29 cells after stimulation is the fastest, HT29 cells is the slowest. Co-culturing two kinds of red and green fluorescent plasmid DNA transfected cells, scratches, we can see the red and green cells evenly distributed. Conlusions TGF-β1 can induce colorectal cancer cells EMT;In colorectal cancer, EMT cells can enhance invasion and metastasis of the non-EMT cells.
出处
《基础医学与临床》
CSCD
北大核心
2014年第9期1165-1170,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(81272758)
