[微小RNA-135b通过靶向腺瘤性息肉病基因对三阴性乳腺癌增殖、侵袭和迁移的影响]
[Effect of miRNA-135b on proliferation, invasion and migration of triple-negative breast cancer by targeting APC].
作者信息
Xin Haina, Jiang Dandan, Lü Zhidong, Sun Suyuan, Kong Jilin, Li Funian
机构信息
Medical College of Qindao Universtty, Breast Disease Diagnosis and Treatment Center, the Affiliated Hospital of Qingdao University, Qingdao 266071, China.
Medical College of Qindao Universtty, Breast Disease Diagnosis and Treatment Center, the Affiliated Hospital of Qingdao University, Qingdao 266071, China; Email:
出版信息
Zhonghua Yi Xue Za Zhi. 2015 Aug 11;95(30):2474-7.
OBJECTIVE
To explore the expression of miRNA-135b in a variety of breast cancer cell lines and its function on the proliferation, invasion and migration in triple-negative breast cancer cell lines by targeting adenomatous polyposis coli (APC).
METHODS
Quantitative real-time (RT)--PCR was used to detect the expression of miRNA-135b in seven breast cancer cell lines and one normal breast cell line. The three-negative breast cancer cell lines MDA-MB-231 and MDA-MB-468 with miRNA-135b high-expression were divided into three groups: the normal growth group, the negative control group (transfected with negative counter-part) and the experimental group (transfected with miRNA-135b inhibitors). RT- PCR and Western blot were used to explore the expression of APC. The proliferation was detected by Cell Counting Kit-8 (CCK-8) method and the invasion and migration were detected by Transwell method.
RESULTS
The levels of miRNA-135b were higher in triple-negative breast cancer cell lines than other types, especially MDA-MB-231 (P=0.001) and MDA-MB-468 (P=0.036). Compared with normal growth group and negative control group, the mRNA and protein of APC were up-regulated in experimental group MDA-MB-231 (P=0.013, P=0.021), MDA-MB-468 (P=0.017, P=0.014). CCK-8 results showed that the proliferation rate of experimental group was decreased compared with negative control and normal growth group MDA-MB-231 (P=0.00 ), MDA-MB-468 (P=0.01). The invasion and migration ability of MDA-MB-231 (P=0.002, P=0.00) and MDA-MB-468 (P=0.01, P=0.00) were obviously decreased after transfected miRNA-135b inhibitors.
CONCLUSION
The expressions of miRNA-135b were higher in most triple-negative breast cancer cell lines than others. miRNA-135b could promote the proliferation, invasion and migration in triple-negative breast cancer cell lines MDA-MB-231 and MDA-MB-468, and APC was one of the target genes of miRNA- 135b by participating in the process of regulation.
目的
探讨微小RNA-135b（miRNA-135b）在多种乳腺癌细胞系中的表达情况，及其通过靶向腺瘤性息肉病基因（APC）对三阴性乳腺癌细胞系增殖、侵袭和迁移的影响。
方法
采用定量实时荧光定量聚合酶链反应（RT-PCR）检测7种乳腺癌细胞系和1种正常乳腺细胞系中miRNA-135b的表达。将miRNA-135b高表达的三阴性乳腺癌细胞系MDA-MB-231和MDA-MB-468分为三组：正常生长组、阴性对照组（转染阴性对照物）和实验组（转染miRNA-135b抑制剂）。采用RT-PCR和蛋白质免疫印迹法检测APC的表达。采用细胞计数试剂盒-8（CCK-8）法检测细胞增殖情况，采用Transwell法检测细胞侵袭和迁移能力。
结果
三阴性乳腺癌细胞系中miRNA-135b的表达水平高于其他类型，尤其是MDA-MB-231（P=0.001）和MDA-MB-468（P=0.036）。与正常生长组和阴性对照组相比，实验组MDA-MB-231（P=0.013，P=0.021）、MDA-MB-468（P=0.017，P=0.014）中APC的mRNA和蛋白表达上调。CCK-8结果显示，实验组的增殖率低于阴性对照组和正常生长组MDA-MB-231（P=0.00）、MDA-MB-468（P=0.01）。转染miRNA-135b抑制剂后，MDA-MB-231（P=0.002，P=0.00）和MDA-MB-468（P=0.01，P=0.00）的侵袭和迁移能力明显降低。
结论
大多数三阴性乳腺癌细胞系中miRNA-135b表达高于其他细胞系。miRNA-135b可促进三阴性乳腺癌细胞系MDA-MB-231和MDA-MB-468的增殖、侵袭和迁移，APC是miRNA-135b参与调控过程的靶基因之一。
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