Department of Laboratory Medicine, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
Key Laboratory of Clinical in vitro Diagnostic Techniques of Zhejiang Province, Hangzhou, China.
Thorac Cancer. 2017 Jul;8(4):304-311. doi: 10.1111/1759-7714.12442. Epub 2017 May 17.
MicroRNA-128 (miR-128) serves as a regulator by inducing cancer cell apoptosis, differentiation, the epithelial-to-mesenchymal transition process, and tumor growth by mediating different targets. NIMA-related kinase 2 (NEK2) is aberrantly expressed in lung cancer. The miR-128/NEK2 pathway has been reported to predict prognosis in colorectal cancer; however, the determination of a relationship between miR-128 and NEK2 in lung cancer has remained elusive. We explored the association between miR-128 and NEK2 in lung cancer.
MiR-128 and NEK2 expression were examined in 15 lung cancer tissues by real time-PCR. Lung cancer SK-MES-1 cells were transfected with miR-128 mimic, an inhibitor or a negative control. MiR-128 and NEK2 expression levels were detected using quantitative real time-PCR and Western blot. SK-MES-1 cell apoptosis was performed by flow cytometry.
Compared to adjacent non-tumor tissues, miR-128 was downregulated and NEK2 was upregulated in 15 lung cancer tissues. Lung cancer SK-MES-1 cells transfected with miR-128 mimic induced a higher apoptotic rate than those transfected with the negative control. Dual luciferase assay further confirmed that NEK2 was a direct target of miR-128 in lung cancer, and transfection with miR-128 mimic could decrease the NEK2 protein level while the miR-128 inhibitor increased NEK2 expression. Finally, the apoptotic effect of lung cancer cells induced by miR-128 mimic could be reversed by NEK2 overexpression.
NEK2 was regulated by miR-128 in lung cancer and miR-128 induced lung cancer cell apoptosis by mediating NEK2 expression.
微小 RNA-128（miR-128）通过介导不同的靶标，诱导癌细胞凋亡、分化、上皮-间质转化过程和肿瘤生长，从而发挥调节作用。有丝分裂后期促进复合物/激酶样蛋白 2（NEK2）在肺癌中异常表达。miR-128/NEK2 通路已被报道可预测结直肠癌的预后；然而，miR-128 与 NEK2 在肺癌中的关系仍未确定。我们探讨了 miR-128 与肺癌中 NEK2 的关系。
通过实时 PCR 检测 15 例肺癌组织中 miR-128 和 NEK2 的表达。用 miR-128 模拟物、抑制剂或阴性对照转染肺癌 SK-MES-1 细胞。通过实时定量 PCR 和 Western blot 检测 miR-128 和 NEK2 的表达水平。通过流式细胞术检测 SK-MES-1 细胞凋亡。
与相邻非肿瘤组织相比，在 15 例肺癌组织中 miR-128 下调，NEK2 上调。转染 miR-128 模拟物的肺癌 SK-MES-1 细胞诱导的凋亡率高于转染阴性对照的细胞。双荧光素酶报告基因实验进一步证实，NEK2 是肺癌中 miR-128 的直接靶标，转染 miR-128 模拟物可降低 NEK2 蛋白水平，而 miR-128 抑制剂则增加 NEK2 表达。最后，miR-128 模拟物诱导的肺癌细胞凋亡效应可被 NEK2 过表达逆转。
NEK2 受 miR-128 调控，miR-128 通过调控 NEK2 表达诱导肺癌细胞凋亡。
