NF-κB抑制剂TBB调控miRNAs影响乳腺癌细胞增殖、侵袭和转移

NF-κB is an important transcription factor and could regulate several downstream genes. It has been evidenced that miRNAs play a critical role in the growth, invasion and metastasis of breast cancer. Our previous study has found that NF-κB inhibitor TBB （4,5,6,7-tetrabromobenzotriazole, TBB） suppresses the proliferation and induces apoptosis of breast cancer cells. Moreover, NF-κB is able to alter the expression of miRNAs. Based on these findings, we hypothesize the theory of "NF-κB-miRNA-downstream genes- growth, invasion and metastasis of breast cancer"..In this research, (1) miRNA transfection and interference methods will be used to determine the impact of differentially expressed miRNAs on the growth, invasion and metastasis of breast cancer. （2）downstream target gene prediction will be conducted using proteomics technology in combination with bioinformatic technology; Luciferase reporter experiment will be performed to confirm the target genes of miRNAs. Taken together, this study will clarify the mechanisms how NF-κB exerts its effect on the growth, invasion and metastasis of breast cancer through regulating the expression of miRNAs and provide a starting point for further research on the role of miRNAs and NF-KB in breast cancer.

NF-κB是一种重要的转录调控因子，miRNAs可影响肿瘤的增殖、侵袭转移等。NF-κB能否通过调控miRNAs影响乳腺癌的发生发展目前还不清楚。我们前期工作发现，NF-κB抑制剂四溴苯三唑（TBB）可抑制乳腺癌细胞增殖、促进凋亡，同时可影响miRNAs表达水平。据此我们提出"NF-κB-miRNA-下游靶基因-乳腺癌细胞增殖、侵袭转移等"假设。.本课题将以前期工作为基础，(1)应用miRNA转染及干扰技术，验证基因芯片筛选出的miRNAs对乳腺癌细胞增殖、侵袭转移的影响，明确受NF-κB调控的miRNAs。(2)采用蛋白质组学技术从蛋白质水平筛选miRNA下游靶基因的新方法，并结合生物学信息技术预测靶基因，荧光报告载体实验验证靶基因。以明确NF-κB通过调控哪些相关miRNAs来影响乳腺癌细胞增殖、侵袭转移。为进一步研究NF-κB及miRNAs在乳腺癌发生、发展中的作用打下初步基础。

NF-κB是一种重要的转录调控因子，miRNAs可影响肿瘤的增殖、侵袭转移等。本研究发现，NF-κB抑制剂四溴苯三唑（TBB）可上调乳腺癌miR-15a和miR-181b水平，抑制乳腺癌细胞增殖和转移。miR-15a水平升高，可使乳腺癌细胞停滞在G1期，同时可抑制细胞迁移能力，但对细胞凋亡无明显影响；进一步研究发现，miR-15a通过其靶基因细胞周期蛋白E1（CCNE1）发挥作用的。上调miR-181b 可抑制乳腺癌细胞增殖和迁移，通过双荧光素酶报告基因系统验证，发现miR-181b通过靶向结合细胞周期蛋白依赖型激酶8（CDK8）发挥作用的。研究表明， “NF-κB—miR-15a—CCNE1—乳腺癌细胞增殖、侵袭转移” 和“NF-κB—miR-181b—CDK8—乳腺癌细胞增殖、侵袭转移”的调控机制在乳腺癌增殖、侵袭和转移中起重要作用，这为进一步研究NF-κB及miRNAs在乳腺癌的作用提供可能的新思路。. miR-135b和 miR-506可调控Hippo-YAP信号通路。高表达miR-135b通过其下游靶基因LATS2，促进乳腺癌细胞增殖和转移； 乳腺癌低水平miR-506可失去对YAP蛋白的靶向调控，使乳腺癌细胞生长加速和凋亡减少。其他如miR-497、miR-26b、miR-195-5p、miR-96、miR-301b、miR-107对乳腺癌细胞增殖、侵袭转移等有一定作用。这为研究miRNAs在乳腺癌的作用提供了一定的依据。