KLF16 suppresses human glioma cell proliferation and tumourigenicity by targeting TFAM
KLF16 suppresses human glioma cell proliferation and tumourigenicity by targeting TFAM
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DOI：
10.1080/21691401.2018.1431654
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发表时间：
2018-01-01
影响因子：
5.8
通讯作者：
Guo, Xieli
中科院分区：
文献类型：
--
作者：
Chen, Xiangrong;Li, Shun;Guo, Xieli
背景资料：方法：采用慢病毒技术将KLF 16或KLF 16-siRNA转染U87 MG细胞，观察KLF 16对U87 MG细胞增殖、分化、凋亡的影响。集落形成实验检测细胞增殖。MTT法检测细胞活力。TUNEL法检测细胞凋亡。流式细胞仪测定细胞周期。进行实时PCR以测试mRNA表达。Western blot检测蛋白水平。荧光素酶法检测KLF 16与线粒体转录因子A（TFAM）的调控关系。染色质免疫沉淀法检测蛋白结合位点。结果：KLF 16在胶质瘤细胞和胶质瘤组织中表达明显降低，在胶质瘤细胞中表达明显降低。KLF 16在体内外均能明显抑制U87 MG细胞的增殖。KLF 16转染降低了与细胞增殖相关的mRNA和蛋白水平。KLF 16靶向TFAM基因转录起始位点附近的一个结合位点，从而抑制胶质瘤细胞的增殖。KLF 16-siRNA表现出相反的影响。结论：KLF 16是直接靶向TFAM的胶质瘤细胞增殖的重要调控因子。
Background: This study aims to via unveiling the novel mechanisms of KLF16 in regulating expression of genes involved in glioma.Methods: KLF16 or KLF16-siRNA was transfected to U87MG cells by lentivirus. Colony formation assay was applied for detecting cell proliferation. MTT assay was adopted to assess cell viability. TUNEL assay was selected to evaluate cell apoptosis. Flow cytometry was used to determine cell cycle. Real-time PCR was performed to test mRNA expression. Western blot was used to detect protein level. Luciferase assay was applied to confirm the regulatory relationship between KLF16 and Mitochondrial transcription factor A (TFAM). Chromatin immunoprecipitation was adopted to test the protein binding site. The nude mouse transplantation tumour experiment was selected to test cancer cell proliferation in vivo.Results: KLF16 was decreased in glioma cells and tissues. KLF16 obviously restrained U87MG cell proliferation both in vivo and in vitro. KLF16 transfection reduced mRNA and protein levels related to cell proliferation. KLF16 targeted a putative binding site near the transcription start sites (TSSs) of TFAM gene, thus suppressing glioma cell proliferation. KLF16-siRNA exhibited the opposite impact. KLF16 presented significant negative correlation with TFAM level in glioma patients.Conclusions: KLF16 is a key regulator of glioma cell proliferation by directly targeting TFAM.