摘要
目的观察TGF—β1能否诱导胃癌KATO—III细胞上皮间质转化及促进胃癌细胞干细胞特性的获得。方法5ng／mETGF-β1处理胃癌KATO-III细胞72h后，置于倒置显微镜下观察其形态学变化，CCK-8法检测TGF-β1对KATO—III细胞增生的影响，RT—PCR与Westernblotting法检测上皮间质转化相关凶子Snail、E—cadherin、N—cadherin，胚胎干细胞相关因子Sox2、OCT4、Nanog、EGFR及肿瘤起始细胞相关标志物CD44、CD133表达的变化。单克隆形成实验研究TGF-β对KATO-1I细胞克隆形成能力的影响。结果TGF-β1处理KATO-Ⅲ细胞后，细胞形态南上皮细胞形态向间质细胞样形态转变；TGF-β1处理组KA—TO—III细胞Snail、N—cadherinmRNA表达相对灰度值为（0．5219±0．0147）、（0．6640±0．0124），显著高于对照组（0．2049±0．0214，P=0．004；0．2722±0．0098，P=0．001），TGF—Bl处理组Snail、N—eadherin蛋白表达相对灰度值为（0．4769±0．0234）、（0．5014±0．0216），显著高于对照组（0．2534±0．0345，P：0．02；0．2026±0．0268，P=0．009），TGF—β1处理组中E—cadherinmRNA及蛋白表达相对灰度值分别为（0．4701±0．0215）、（0．1349±0．0258），昆著低于对照组（0．6792±0．0157，P=0．01；0．6055±0．0227，P=0．004）：TGF—β1处理后KATO—III细胞增生能力明显降低（P〈0．05）。与对照组相比（0．143±0．013、0．156±0．025、0．325±0．046），胚胎干细胞相关凶子Sox2、OCT4、NanogmRNA表达相对灰度值显著增加（0．594±0．039，P=0．001；0．438±0．033，P=0．001；0．489±0．037，P=0．03），TGF—β1处理组CIN4与CDl33mRNA表达相对灰度值分别为（0．437±0．037）与（0．543±0．028），显著高于对照组（0．247±0．024，P=0．000：0．139±0．016，P=0．000）；TGF—β1处理组CD44与CD133蛋白表达相对灰度值分别为（0．429±0．034）与（0．316±0．027），显著高于对照组（0．152
Objective To investigate if TGF- β1 induces epithelial- mesenchymal transition （EMT） and promotes the obtaining of sternness characteristics in gastric cancer cell lines. Methods After KATO-III cells were cultured with or without 5 ng/mL TGF-β1, the morphological change was observed and compared under phase-contrast microscopy. At the same time, the effect of TGF- β1 on the proliferation of KATO- III ceils was detected by CCK-8. On the other hand, the mRNA and protein＇ s expressions of EMT-related factors, ESC markers and TICs markers were analyzed by RT-PCR and Western blotting methods too. Results TGF-131 induced morphological alterations from epithelial to mesenchymal cells. The proliferation of KATO- m cells was inhibited after treated with TGF-β1 （P 〈 0. 05）. After treated with TGF-β1, the relative mRNA expression levels of Snail （0.5219 ± 0.0147 ） and N- cadherin （0. 6640 ± 0.0124） were higher than that in control group （0. 2049 ± 0.0214, P = 0. 004, 0. 2722 ± 0. 0098, P = 0. 001 ）, the relative protein expression levels of Snail （0. 4769 ± 0. 0234） and N- cadherin （0. 5014 ± 0. 0216） were higher than that in control group （0. 2534 ± 0. 0345, P = 0. 02, 0. 2026 ± 0. 0268, P = 0.009）, while the relative E-cadherin mRNA and protein levels in TGF-β1 treated group （0. 4701 ±0. 0215, 0. 1349 ± 0. 0258） were lower than that in control group （0. 6792 ± 0. 0157, P = 0. 01 ; 0. 6055 ±0. 0227, P = 0.004）, while the relative mRNA expressions of ESC markers such as Sox2, OCT4, Nanog in TGF-β1 treated group （0. 594±0. 039,0. 438 ± 0. 033,0. 489 ± 0. 037 ） were higher than that in control group （0. 143±0. 013, P = 0. 001, 0. 156 ± 0. 025, P = 0. 001,0. 325 ± 0. 046, P = 0.03 ） , the relative mRNA expression levels of CD44 （0. 437 ± 0. 037 ） and CD133 （0. 543 ± 0. 028 ） were higher than that in control group （0. 247 4-0. 024 ,P = 0. 000, 0. 139 ±-0. 016 ,P = 0. 000 ） , the relative protein expression levels of CD44 （0. 429 ± 0.
出处
《国际外科学杂志》
2012年第12期824-829,F0003,共7页
International Journal of Surgery
基金
国家自然科学基金（No.81101850）
上海市卫生局重点课题（No.2010018）
上海市科学技术委员会基金（No.09411962300）
上海市教育委员会基金（No.jdy10022）
