miR-149上调E-cadherin抑制乳腺癌细胞侵袭、迁移

Up-regulation of miR-149 by E-cadherin inhibits breast cancer cell invasion and migration

目的:探讨微小RNA(miR)-149在乳腺癌中的表达及过表达对乳腺癌细胞侵袭、迁移的影响和可能的作用机制。方法:Realtime-PCR检测60例乳腺癌组织、癌旁组织和50例正常乳腺组织miR-149相对表达量。miR-149mimics瞬时转染乳腺癌MDA-MB-231细胞(miR-149转染组),并设置空白载体转染组和培养液添加组,Realtime-PCR检测各组细胞中miR-149相对表达量,transwell小室法检测各组细胞侵袭和迁移能力,western-blot检测E-cadherin蛋白表达量。结果:乳腺癌组织miR-149相对表达量显著低于癌旁组织和正常乳腺组织,差异有统计学意义(P<0.05);而癌旁组织与正常乳腺组织miR-149相对表达量比较差异无统计学意义(P>0.05)。miR-149转染组转染后不同时间点miR-149表达量显著高于空白载体转染组和培养液添加组,差异有统计学意义(P<0.05);空白载体转染组和培养液添加组比较差异均无统计学意义(P>0.05)。miR-149转染组侵袭细胞数和迁移细胞数均显著低于空白载体转染组和培养液添加组,差异有统计学意义(P<0.05);空白载体转染组和培养液添加组比较差异均无统计学意义(P>0.05)。miR-149转染组E-cadherin蛋白表达显著高于空白质粒转染组和培养液添加组,差异有统计学意义(P<0.05);空白载体转染组和培养液添加组比较差异均无统计学意义(P>0.05)。结论:乳腺癌组织中miR-149低表达,过表达miR-149可能通过上调E-cadherin抑制乳腺癌细胞侵袭、迁移。

Objective： To investigate the effects and possible mechanism of MicroRNA（ miR）-149 expression and over-expression in breast cancer cells on their invasion and migration. Methods： Relative transcript levels of miR-149 in 60 cases of breast cancer tissues and para-carcinoma tissues,50 cases of normal breast tissues were detected by real-time PCR. Transient transfection of breast cancer MDA-MB-231 was conducted by miR-149 mimics（ MiR-149 transfection group）,transient transfection by empty vector was set as control group and culture solution group was set,relative transcript levels of miR-149 in each group were detected by realtime-PCR,cell invasion and migration abilities in each group were detected by transwell chambers method,protein expression of Ecadherin was detected by western-blot. Results： Relative transcript levels of miR-149 in breast cancer tissues were significantly lowerthan that in para-carcinoma tissues and normal breast tissue（ P〈0. 05）. Relative transcript levels of miR-149 in para-carcinoma tissues and normal breast tissue were not statistically significant（ P〈0. 05）. Relative transcript levels of miR-149 in different time points after transfection（ MiR-149 transfection group） were significantly higher than that in transient transfection by empty vector and culture solution group（ P〈0. 05）. Difference of control group and culture solution group was not statistically significant（ P〈0. 05）. Invasive cell number and migration cell number in miR-149 transfection group were both significantly lower than that in control group and culture solution group（ P〈0. 05）. Difference of control group and culture solution group was not statistically significant（ P〈0. 05）. Protein expression of E-cadherin in miR-149 transfection group was significantly higher than that in control group and culture solution group（ P〈0. 05）. Difference of control group and culture solution group was not statistically significant（ P〈0. 05）. Conclusions： miR-149 is low expression in breast cancer tissues,over-expression of miR-149 may inhibit breast cancer cell invasion and migration by up-regulation of E-cadherin.