C-藻蓝蛋白抑制TGF-β1诱导的宫颈癌Caski细胞上皮-间充质转化  

C-phycocyanin inhibits epithelial-mesenchymal transformation of TGF-β1-induced cervical cancer Caski cells

作　　者：姬欢欢 董晓雷 朱峰 刘国祥 韩晶晶 杨方浩 杨一凡 李冰 JI Huanhuan;DONG Xiaolei;ZHU Feng;LIU Guoxiang;HAN Jingjing;YANG Fanghao;YANG Yifan;LI Bing(Department of Biology,Faculty of Medicine,Qingdao University,Qingdao 266021,Shandong,China)

机构地区：[1]青岛大学医学部基础医学院遗传学与细胞生物学系,山东青岛266021

出　　处：《中国肿瘤生物治疗杂志》2020年第2期129-134,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81871231,81471546,81001346);山东省高等学校青年创新科技计划资助(No.2019KJK016);青岛市民生科技计划项目资助(No.18-6-1-91-nsh)。

摘　　要：目的:探究C-藻蓝蛋白(C-phycocyanin,C-PC)对转化生长因子β1(transforming growth factor beta 1,TGF-β1)诱导的宫颈癌Caski细胞上皮-间充质转化(epithelial-mesenchymal transition, EMT)的影响。方法:根据处理方法不同将Caski细胞分为3组即10 ng/ml TGF-β1处理组、10 ng/ml TGF-β1+300μg/ml C-PC共处理组和对照组(未加药处理),处理24 h后观察细胞的形态变化;采用划痕实验及Transwell实验分别检测TGF-β1和C-PC对Caski细胞迁移和侵袭的影响;Western blotting方法检测C-PC对TGF-β1诱导的Caski细胞上皮表型标志蛋白E-cadherin、间质表型标志蛋白N-cadherin表达水平的影响;qPCR法检测间质表型相关锌指转录因子Snail、Zeb1、Twist m RNA的表达。结果:TGF-β1处理组Caski细胞失去原有的上皮表型的特征,而TGF-β1+C-PC共处理组细胞保持正常的上皮表型的特征;TGF-β1处理组Caski细胞的迁移率[(60.0±1.4)%vs(33.5±2.2)%、(40.0±2.8)%,均P<0.05]和侵袭穿膜细胞数[(108.2±6.2)vs(25.2±3.1)、(39.8±5.4)个,均P<0.01]较共处理组和对照组显著升高;与对照组相比,TGF-β1处理组Caski细胞中E-cadherin表达显著降低(P<0.05),Twist、Snail、Zeb1的mRNA表达水平显著升高(P<0.01),而加入C-PC共处理可逆转上述改变(P<0.05或P<0.01),同时显著降低N-cadherin的蛋白表达水平(均P<0.05)。结论:C-PC处理能够抑制TGF-β1诱导的Caski细胞侵袭转移能力进而影响EMT过程,其机制可能与C-PC处理降低Twist、Snail、Zeb1的mRNA表达有关。Objective: To investigate the effect of C-phycocyanin(C-PC) on the epithelial-mesenchymal transition(EMT) of cervical cancer Caski cells induced by transforming growth factor beta1(TGF-β1). Methods: According to different treatment methods, Caski cells were divided into three groups: 10 ng/ml TGF-β1 treatment group, 10 ng/ml TGF-β1+300 μg/ml C-PC co-treatment group and control group(untreated). After 24 h of treatment, the morphological changes of Caski cells were observed, and the effects of TGF-β1 and C-PC on the migration and invasion of Caski cells were detected by Scratch test and Transwell test, respectively. Western blotting was used to detect the effect of C-PC on the expression of epithelial phenotypic marker protein E-cadherin and stromal phenotypic marker protein N-cadherin in TGF-β1-induced Caski cells, and qPCR was used to detect the mRNA expressions of EMT related factors Snail,Zeb1 and Twist. Results: Caski cells in the TGF-β1 treatment group lost the characteristics of the original epithelial phenotype, while the cells in the TGF-β1+C-PC co-treatment group maintained the characteristics of normal epithelial phenotype;the migration rate([60.0±1.4]% vs [33.5±2.2]%, [40.0±2.8]%, both P<0.05) and the number of invasive transmembrane cells([108.2±6.2] vs [25.2±3.1],[39.8±5.4], both P<0.01]) of Caski cells in the TGF-β1 treatment group were significantly higher than those in the co-treatment group and the control group. Compared with the control group, the expression of E-cadherin in Caski cells treated with TGF-β1 decreased significantly(P<0.05), while the mRNA expressions of Twist, Snail and Zeb1 increased significantly(all P<0.05);However, co-treatment with C-PC reversed above changes(P<0.05 or P<0.01), and significantly decreased the protein expression level of N-cadherin(P<0.05). Conclusion: C-PC treatment can inhibit the invasion and metastasis ability of Caski cells induced by TGF-β1 and further affects the EMT process. The mechanism may be related to the decrease of m RNA expres

关 键 词：C-藻蓝蛋白 TGF-Β1 宫颈癌 迁移 侵袭 CASKI细胞 

分 类 号：R737.33[医药卫生—肿瘤] R730.54[医药卫生—临床医学]