CAJ | 학술논문

目的观察微小RNA (miR)-30a对乳腺癌细胞增殖、侵袭能力的影响,并探讨其与锌指结构E-box同源结合框2(ZEB2)的关系.方法采用实时定量聚合酶链反应(Real-time PCR)技术检测乳腺癌组织及癌旁组织中miR-30a的表达.用miR-30a的模拟物片段(has-miR-30a mimics)瞬时转染乳腺癌细胞MCF-7及MDA-MB-231.通过细胞增殖检测试剂盒(CCK-8)检测miR-30a对细胞增殖能力的影响.通过小室(Transwell)检测miR-30a对乳腺癌细胞侵袭能力的影响.通过TargetScan软件预测miR-30a的潜在靶基因为ZEB2,并利用荧光素酶报告基因实验和Western blot法证实miR-30a直接靶向靶基因ZEB2.结果乳腺癌组织与癌旁组织的miR-30a相对表达水平的比值为0.47 ±0.01(P＜0.05).在MCF-7细胞和MDA-MB-231细胞中过表达miR-30a,培养48 h后的吸光度值分别为2.03±0.19、2.56±0.21,低于对照组3.22±0.07、3.49±0.19,细胞侵袭抑制率分别为(36.04±0.03)％、(53.61±0.03)％,ZEB2的蛋白表达水平分别下调了40％和60％(P＜0.05).荧光素酶报告基因实验证实ZEB2是miR-30a的直接靶基因.结论 miR-30a在乳腺癌组织和细胞中的表达明显下调,miR-30a可以通过靶向ZEB2抑制乳腺癌细胞的增殖和侵袭.
목적 관찰미소RNA (miR)-30a대유선암세포증식、침습능력적영향,병탐토기여자지결구E-box동원결합광2(ZEB2)적관계.방법 채용실시정량취합매련반응(Real-time PCR)기술검측유선암조직급암방조직중miR-30a적표체.용miR-30a적모의물편단(has-miR-30a mimics)순시전염유선암세포MCF-7급MDA-MB-231.통과세포증식검측시제합(CCK-8)검측miR-30a대세포증식능력적영향.통과소실(Transwell)검측miR-30a대유선암세포침습능력적영향.통과TargetScan연건예측miR-30a적잠재파기인위ZEB2,병이용형광소매보고기인실험화Western blot법증실miR-30a직접파향파기인ZEB2.결과 유선암조직여암방조직적miR-30a상대표체수평적비치위0.47 ±0.01(P＜0.05).재MCF-7세포화MDA-MB-231세포중과표체miR-30a,배양48 h후적흡광도치분별위2.03±0.19、2.56±0.21,저우대조조3.22±0.07、3.49±0.19,세포침습억제솔분별위(36.04±0.03)％、(53.61±0.03)％,ZEB2적단백표체수평분별하조료40％화60％(P＜0.05).형광소매보고기인실험증실ZEB2시miR-30a적직접파기인.결론 miR-30a재유선암조직화세포중적표체명현하조,miR-30a가이통과파향ZEB2억제유선암세포적증식화침습.
Objective To investigate the expression of microRNA (miRNA,miR)-30a in breast cancer and its effect on proliferation and invasion of breast cancer cells,as well to evaluate the relationship of zinc-finger E-box binding homeobox 2 (ZEB2) and miR-30a.Methods The expression of miR-30a in 30 cases of breast tumor tissues and corresponding paracancerous tissues was detected by real-time quantitative polymerase chain reaction (Real-time PCR).Has-miR-30a mimics was transfected into MCF-7 cells and MDA-MB-231 cells.The effect of miR-30a on proliferation of cells was detected by cell counting kit-8 (CCK-8) assay.The invasion of cells was detected by Transwell assay.The potential targets of miR-30a were predicted by TargetScan and confirmed by luciferase reporter assay and Western blotting.Results The relative expression rate of miR-30a in breast cancer tissues and corresponding paracancerous tissues was 0.47 ±0.01 (P ＜0.05).The cell absorbance values were (2.03 ± 0.19) and (2.56 ±0.21) after 48 h cell culture,which were lower than the control [(3.22 ±0.07) and (3.49 ± 0.19)].The invasion inhibition rate was (36.04 ± 0.03) ％ and (53.61 ± 0.03) ％.The protein expression of ZEB2 was downregulated by 40％ and 60％ respectively.The luciferase reporter assay showed that ZEB2 was a direct target of miR-30a.Conclusion MiR-30a is significantly downregulated in breast cancer tissues and cells,and can inhibit cell proliferation and invasion through targeting ZEB2.