TGF-β1对SKOV-3细胞EMT的影响及相关分子机制研究  

Effects of TGF-β1 on EMT of SKOV-3 cells and its molecular mechanism

作　　者：闫晓楠[1] 杜辉[2] 左宏玲[2] 张娜[2] 王惠兰[2] 徐春琳[2] YAN Xiaonan;DU Hui;ZUO Hongling(Department of Ultrasonic Diagnosis and Treatment in Obsterics and Gynecology,The Second Hospital of Hebei Medical University,Hebei,Shijiazhuang 050000,China;不详)

机构地区：[1]河北医科大学第二医院妇产超声诊疗科,石家庄市050000 [2]河北医科大学第二医院妇科,石家庄市050000

出　　处：《河北医药》2021年第6期833-836,841,共5页Hebei Medical Journal

摘　　要：目的研究TGF-β1(TGF-β1)对上皮性卵巢癌SKOV-3细胞上皮间质转化(EMT)的影响及相关分子机制。方法MTT法检测各浓度TGF-β1(1、2、5以及10 ng/ml)干预24、48 h后,SKOV-3细胞增殖所受影响(对照组TGF-β10 ng/ml);Transwell检测培养48 h后SKOV-3细胞的侵袭能力,实验分组:TGF-β1组(浓度1、2、5、10 ng/ml)、LY294002组(PI3K抑制剂,浓度10μmol/ml)、LY294002+TGF-β1组(浓度10μmol/ml+10 ng/ml)、对照组(PBS);蛋白印迹(Western blot)法检测培养48 h后各组丝氨酸-苏氨酸蛋白激酶B(Serine-threonine protein kinase B,p-Akt)、转录因子Twist(Transcriptional factor Twist,Twist)、神经钙黏(Nerual-cadherin,N-cadherin)蛋白及上皮钙黏(Epithelial-cadherin,E-cadherin)蛋白等的表达情况,分组如下:TGF-β1组(浓度10 ng/ml)、LY294002组(浓度10μmol/ml)、LY294002+TGF-β1组(浓度10μmol/ml+10 ng/ml)、对照组(RPMI-1640)。结果MTT:TGF-β1对SKOV-3细胞无明显促增殖作用(P>0.05);Transwell:不同浓度TGF-β1干预后SKOV-3穿过小室细胞数目多于对照组,呈浓度依赖性(P<0.05);LY294002单独或联合TGF-β1组,SKOV-3细胞运动侵袭能力均显著降低(P<0.05);Western blot:经TGF-β1干预后p-Akt、Twist、N-cadherin蛋白表达水平显著增加,E-cadherin蛋白表达水平显著降低;LY294002单独或联合TGF-β1可逆转上述蛋白的表达情况(P<0.05)。结论TGF-β1可对PI3K/Akt信号通路起激活作用,从而诱导SKOV-3细胞出现EMT,促使其侵袭转移能力增强。Objective To investigate the effect of TGF-β1 on epithelial mesenchymal transition(EMT)of epithelial ovarian cancer cell line SKOV-3,and to explore related molecular mechanism.Methods After SKOV-3 cells were treated by different concentration of TGF-β1 for 24-hour and 48-hour,who were divided into TGF-β1 group(1,2,5,10ng/ml),LY294002 group(PI3K inhibitor,10μmol/ml),LY294002+TGFβ1 group(10μmol/ml+10ng/ml),and control group(PBS).Western Blot was used to detect expression levels of serine threonine protein kinase B(p-Akt),transcription factor twist(Twist),epithelial cadherin(E-cadherin)protein and nervous cadherin(N-cadherin)after 48-hour treatment.Results The MTT experiment results showed that TGF-β1 had no significant promotive effects on the proliferation of SKOV 3 cells(P>0.05).The Transwell results showed that the numbers of SKOV-3 cells that passed through the chamber were significantly more than those in control group after TGF-β1 intervention,with a concentration dependent manner(P<0.05).The motility and invasion ability of cells in simple LY294002 or combination with TGF-β1 group were significantly decreased(P<0.05).Western Blot results showed that after TGF-β1 intervention the expression levels of p-Akt,twist and N cadherin protein were significantly increased,however the expression levels of E cadherin were significantly decreased.Moreover simple LY294002 or combination with TGF-β1 could reverse the expression changes of the above proteins(P<0.05).Conclusion TGF-β1 can promote EMT of SKOV-3 cells by activating PI3K/Akt signaling pathway,so as to enhance the invasion and metastasis abilities of tumor cells.

关 键 词：上皮性卵巢癌 上皮间质转化 TGF-Β1 细胞增殖 侵袭转移 

分 类 号：R737.31[医药卫生—肿瘤]