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纸质出版:2014
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杜俊泽,范林军,齐晓伟,孙鹏,王明浩,明佳,姜军.MicroRNA-206可能通过调控Cx43表达调节人乳腺癌细胞的转移能力[J].陆军军医大学学报(原第三军医大学学报),2014,36(02):115-120.
Du Junze,Fan Linjun,Qi Xiaowei,Sun Peng,Wang Minghao,Ming Jia,Jiang Jun.Inhibitory effects of microRNA-206 on migration and invasion of breast cancer by targeting Cx43[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(02):115-120.
杜俊泽,范林军,齐晓伟,孙鹏,王明浩,明佳,姜军.MicroRNA-206可能通过调控Cx43表达调节人乳腺癌细胞的转移能力[J].陆军军医大学学报(原第三军医大学学报),2014,36(02):115-120. DOI:
Du Junze,Fan Linjun,Qi Xiaowei,Sun Peng,Wang Minghao,Ming Jia,Jiang Jun.Inhibitory effects of microRNA-206 on migration and invasion of breast cancer by targeting Cx43[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(02):115-120. DOI:
目的 通过干预人乳腺癌细胞株MCF-7和MDA-MB-231中微小RNA206(microRNA-206
miR-206)的表达,观察连接蛋白43(connexin43,Cx43)的表达以及细胞在增殖、迁移和侵袭能力等方面的变化。 方法 使用荧光定量RT-PCR(qRT-PCR)检测正常乳腺细胞MCF-10A和MCF-7、MDA-MB-231细胞中miR-206的表达水平。通过构建慢病毒载体转染MCF-7及MDA-MB-231细胞,分别升高和降低miR-206的表达水平;采用qRT-PCR和Western blot法检测转染后Cx43的mRNA及蛋白表达水平的变化。利用CCK-8法检测细胞增殖能力,Transwell小室法检测细胞迁移和侵袭能力。 结果 MCF-7和MDA-MB-231中miR-206的表达均明显低于正常细胞株;其中MCF-7细胞中miR-206表达水平较MDA-MB-231中低,Cx43表达水平较MDA-MB-231中高。转染慢病毒包装靶向增强miR-206表达的shRNA载体LV-hsa-miR-206(1569-11)后,MCF-7细胞的miR-206 表达明显上升,Cx43的mRNA表达水平及蛋白表达水平均明显下降;细胞的增殖、迁移和侵袭能力明显降低。转染慢病毒包装靶向抑制miR-206表达的shRNA载体LV-hsa-miR-206-inhibition后,MDA-MB-231细胞的miR-206 表达明显下降,Cx43 mRNA表达水平及蛋白表达水平均明显上升;细胞的增殖、侵袭和迁移能力明显增强;以上差异均具有统计学意义(
P
0.05)。 结论 干预miR-206的表达可导致Cx43表达水平的变化,并影响人乳腺癌细胞MCF-7、MDA-MB-231的增殖、迁移和侵袭能力;miR-206可能通过抑制Cx43的表达调节了乳腺癌细胞的转移能力。
Objective To explore the roles of microRNA-206 (miR-206) and connexin 43 (Cx43) in the cell proliferation
migration and invasion of breast cancer. Methods With qRT-PCR
the miR-206 expression levels of breast cancer cell lines MDA-MB
-231 and MCF-7 and normal mammary gland cell line MCF-10A were detected. The levels of miR-206 in MDA-MB-231 and MCF-7 cells were up-regulated or down-regulated by lentiviral vector-mediated transfection. Cx43 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Results It was found that there was obviously lower expression of miR-206 in breast cancer cells than in normal cells. MCF-7 cells showed lower miR-206 level but higher Cx43 level than MDA-MB-231 cells. After shRNA-mediated miR-206 up-regulation
MCF-7 cells displayed increase of miR-206 level
decrease of Cx43 mRNA and protein expression
and suppression of cell proliferation
migration and invasion (
P
0.05). After shRNA-mediated miR-206 down-regulation
MDA-MB-231 cells displayed decrease of miR-206 level
increase of Cx43 mRNA and protein expression
and enhancement of cell proliferation
migration and invasion (
P
0.05). Conclusion MicroRNA-206 inhibits migration and invasion of breast cancer cells through down-regulation of Cx43 expression possibly
which indicates the tumor-suppressive function of microRNA-206 in breast cancer.
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