CD44在肿瘤细胞上皮-间充质转化及其侵袭过程中的作用  

The epithelial -mesenchymal transition induced by CD44 gene prompted ovarian cancer cell metastasis

作　　者：韩丽丽[1] 张玲玲[2] 杨蕊蕊[1] 

机构地区：[1]滨州市人民医院妇科,山东256610 [2]滨州市人民医院肿瘤科

出　　处：《中华实验外科杂志》2015年第12期3135-3138,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨CD44基因对人卵巢上皮性癌（EOC）细胞上皮一间充质转化和转移能力的影响及其机制。方法培养EOC细胞SK—OV3，取对数生长期细胞用于实验。采用基因共转染法，将细胞设为两组，CD44组用PEFCD44质粒和PSV2neo共转染（作用48、72h），阴性对照组用PEF和PSV2neo共转染。采用划痕实验观察细胞迁移能力，倒置显微镜观察细胞形态，实时定量聚合酶链反应（Real-timePCR）检测Snail、SlugmRNA表达水平，Westernblot检测E-钙黏蛋白（E—cadherin）、N-钙黏蛋白（N-cadherin）和波形蛋白（Vimentin）蛋白表达水平。结果划痕24、48h时，CD44组剩余划痕面积分别为（0．37±0．08）％和（0．16±0．03）％，均显著小于对照组的（0．58±0．13）％和（0．49±0．11）％，差异有统计学意义（P〈0．05）。与对照组比较，CIM4组转染SK—OV3细胞72h后，细胞由上皮形态转变为间质形态，即细胞由立方形或椭圆形转变为纺锤形或多角形，且细胞间连接松散。CD44作用于SK—OV3细胞48h后，可显著上调Snail、SlugmRNA表达水平，Snail、SlugmRNA的相对表达量为1．573±0．126和2．218±0．252，均显著高于对照组的1．024±0．079和0．915±0．057，差异有统计学意义（P〈0．05）；CD44转染于SK-OV3细胞72h，E-cadherin蛋白表达水平较对照组明显减少[（0．40±0．02）ng／L]，与对照组比较[（0．80±0．05）ug／L]，差异有统计学意义（P〈0．05）；CD44转染于SK—OV3细胞72h可使间质细胞标志物N—eadherin与Vimentin蛋白表达水平显著增高[（1．08±0．06）ng／L和（1．26±0．08）ng／L]，与对照组比较[（0．51±0．03）ng／L和（0．61±0．03）ns／L]，差异均有统计学意义（P〈0．05）。结论CD44可通过抑制E—cadherin蛋白的表达，诱导EOC细胞发生上皮-间充质转化，进而增强EOC细胞的转移能力。Objective To explore the effect of CD44 gene on human epithehal ovarian cancer （EOC） cell metastasis. Methods EOC cells line SK - OV3 were located in 2 groups. The CIM4 group was transfected with PEF CD44 plasmid and PSV2neo （48 h or 72 h）, and the negative control group was transfected with PEF and PSV2neo. The effect of CD44 on cell metastasis was observed by the scratch as- say, morphological change was observed by phase contrast microscope, the expression of Snail and Slug mRNA was detected by real- time quantitative polymerase chain reaction （Real- time PCR）, the expres- sion of E - cadherin, N - cadherin and Vimentin protein was detected by Western blotting. Results After scratching for 24 h and 48 h, the remaining wound area rates of CD44 group were （0. 37 ± 0. 08） % and （0. 16 ± 0. 03 ） %, were significantly less than control group [ （0. 58 ± 0. 13 ） % and （ 0.49 ± 0. 11 ） %, P 〈 0. 05 ]. Treated with PEF CIM4 for 72 h, the morphology of SK - OV3 cell in CD44 group changed from epithelial cuboidal or cobbly morphology to spindle - shaped or polygonal morphology, and increased intercellular separation. The expression of Snail and Slug mRNA of SK - OV3 cell in CD44 group were sig- nificantly up - regulated after treated with CIM4 for 48 h （ 1. 573 ±0. 126 and 2. 218 ±0. 252 vs. 1. 024 ± 0. 079 and 0. 915 ± 0. 057, P 〈 0.05 ]. Treatment of PEF CIM4 for 72 h could down - regulate the E - cadherin protein, meanwhile up - regulate the N - cadherin and Vimentin protein. Compared with control group, there was significant difference, respectively [ （0.40 ± 0. 02 ） ng/L, （ 1.08 ± 0. 06 ） ng/L and （1.26±0.08） ng/Lvs. （0.80±0.05） ng/L, （0.51 ±0.03） ng/L and （0.61 ±0.03） ng/L, P〈 0. 05 ]. Conclusion CDdd prompted the metastasis of EOC cell through inducing the epithelial - mesen- chymal transition.

关 键 词：卵巢癌 CD44基因 上皮-间充质转化 

分 类 号：R735.340.2[医药卫生—肿瘤]