miR-124通过靶向调控AURKA抑制胶质瘤细胞的增殖

目的:探索miR-124及AURKA对胶质瘤细胞增殖能力的影响并探索其机制.方法:通过PCR测定胶质瘤组织及非肿瘤组织中miR-124、AURKA的表达水平;将携带miR-124的脂质体miR-124 mimic、空载体miR-124转染入胶质瘤细胞系LN-229和T98G中,吸光度观察miR-124对胶质瘤细胞系LN...     

Issues in interpreting the in vivo activity of Aurora-A

Introduction: Based on its role as a mitotic regulatory kinase, overexpressed and associated with aneuploidy in cancer, small-molecule inhibitors have been developed for Aurora-A (AURKA) kinase. In preclinical and clinical assessments, these agents have shown efficacy in inducing stable disease or therapeutic response. In optimizing the use of Aurora-A inhibitors, it is critical to have robust capacity to measure the kinase activity of Aurora-A in tumors.

Areas covered: We provide an overview of molecular mechanisms of mitotic and non-mitotic activation of Aurora-A kinase, and interaction of Aurora-A with its regulatory partners. Typically, Aurora-A activity is measured by use of phospho-antibodies targeting an autophosphorylated T288 epitope. However, recent studies have identified alternative means of Aurora-A activation control, including allosteric regulation by partners, phosphorylation on alternative activating residues (S51, S98), dephosphorylation on inhibitory sites (S342) and T288 phosphorylation by alternative kinases such as Pak enzymes. Additional work has shown that the relative abundance of Aurora-A partners can affect the activity of Aurora-A inhibitors, and that Aurora-A activation also occurs in interphase cells.

Expert opinion: Taken together, this work suggests the need for comprehensive analysis of Aurora-A activity and expression of Aurora-A partners in order to stratify patients for likely therapeutic response.     

The role of AURKA/miR‐199b‐3p in hepatocellular carcinoma cells

BackgroundPrevious studies proved that AURKA functions as an oncogene in several cancers. This article aimed to probe the miRNA‐induced regulatory mechanism of AURKA in hepatocellular carcinoma (HCC).MethodsDifferentially expressed genes in TCGA‐LIHC dataset were screened by bioinformatics methods. Levels of miR‐199b‐3p and AURKA mRNA were examined by qRT‐PCR. Western blot was utilized to evaluate protein levels of AURKA, p‐AKT, and AKT. Dual‐luciferase assay was introduced to explore their interaction. MTT, colony formation, scratch healing, transwell, and flow cytometry assays were introduced into cell proliferation, migration, invasion, and apoptosis assessment. The impact of miR‐199b‐3p/AURKA axis on HCC tumor growth was determined in a tumor xenograft model.ResultsWe found that AURKA was highly expressed in HCC and was coupled to poor prognosis of HCC. As manifested by cellular assays, compared to the normal cells HL‐7702, AURKA presented notably high expression in HCC cell lines. Overexpressed AURKA evidently impelled the proliferation, colony formation, migration, and invasion of HCC cells while suppressing apoptosis. The regulatory gene upstream of AURKA was predicted to be miR‐199b‐3p by bioinformatics method, and there was a markedly negative correlation between the two. Overexpressed miR‐199b‐3p constrained HCC cell proliferation, migration, and invasion while fostering apoptosis, which could be counteracted by upregulating AURKA. MiR‐199b‐3p repressed the tumor growth in vivo by targeting AURKA and affected PI3K/AKT signaling pathway.ConclusionTo summarize, this study implied the regulatory mechanism of miR‐199b‐3p/AURKA axis in HCC, and supplied optional therapeutic targets for HCC patients.     

Genetic polymorphisms in AURKA and BRCA1 are associated with breast cancer susceptibility in a Chinese Han population

Centrosome defects can result in aneuploidy and genomic instability, and have important implications for breast cancer development. The Aurora-A and BRCA1 proteins interact and both are strongly involved in centrosome regulation. Genetic variants in these two genes may have an effect on breast cancer development. Here, we report a comprehensive single nucleotide polymorphism (SNP) and haplotype-tagging association study on these two genes in 1334 breast cancer cases and 1568 unaffected controls among the Chinese Han population. Apart from a missense SNP, rs2273535 (Phe31Ile), and a probable risk SNP, rs2064863, six htSNPs were analysed in three high-LD blocks of AURKA spanning from 10 kb upstream to 2 kb downstream of AURKA. For BRCA1, six htSNPs were analysed in a large high-LD region covering 98 kb (10 kb was extended to each end of BRCA1). The results showed that four SNPs in AURKA (data in recessive model, rs2273535: OR = 2.19, 95% CI = 1.03-4.66, p = 0.0422; rs2298016: OR = 0.38, 95% CI = 0.18-0.82, p = 0.0141; rs6024836: OR = 1.54, 95% CI = 1.18-2.00, p = 0.0014; rs10485805: OR = 0.68, 95% CI = 0.47-0.98, p = 0.0380) and one SNP in BRCA1 (rs3737559, dominant model OR = 1.35, 95% CI = 1.11-1.64, p = 0.0030) were associated with breast cancer susceptibility. After correction for multiple comparisons (FDR = 0.05), only rs6024836 and rs3737559 remained significant. Two haplotypes (CC of block 2, OR = 20.74, 95% CI = 4.35-98.88, p = 0.0001; GG of block 3, OR = 1.32, 95% CI = 1.12-1.56, p = 0.0010) and one diplotype (AG-GG of block 3, OR = 1.63, 95% CI = 1.18-2.26, p = 0.0031) within AURKA showed strong associations with breast cancer risk. One haplotype of BRCA1 (CTGTTG, OR = 1.30, 95% CI = 1.06-1.59, p = 0.0118) was also associated with breast cancer risk. However, women harbouring both at-risk genotypes of Aurora-A and BRCA1 were at a slightly increased risk compared with those harbouring either at-risk variant alone. Common genetic variants in the AURKA and BRCA1 genes may contribute to breast cancer development.     

Gossypin inhibits gastric cancer growth by direct targeting of AURKA and RSK2

Gossypin is a flavone extracted from Hibiscus vitifolius, which has been reported to exhibit anti‐inflammatory, antioxidant, and anticancer activities. However, the anticancer properties of gossypin and its molecular mechanism of action against gastric cancer have not been fully investigated. In the present study, we report that gossypin is an Aurora kinase A (AURKA) and RSK2 inhibitor that suppresses gastric cancer growth. Gossypin attenuated anchorage‐dependent and anchorage‐independent gastric cancer cell growth as well as cell migration. Based on the results of in vitro screening and cell‐based assays, gossypin directly binds to and inhibits AURKA and RSK2 activities and their downstream signaling proteins. Gossypin decreased S phase and increased G2/M phase cell cycle arrest by reducing the expression of cyclin A2 and cyclin B1 and the phosphorylation of the CDC protein. Additionally, gossypin also induced intrinsic apoptosis by activating caspases and PARP and increasing the expression of cytochrome c. Our results demonstrate that gossypin is an AURKA and RSK2 inhibitor that could be useful for treating gastric cancer.     

AURKA基因在口腔鳞癌中的表达及意义

目的 检测极光激酶A(AURKA)基因在口腔鳞癌中的表达及意义.方法 收集20例口腔正常组织及42例口腔鳞状细胞癌(OSCC)组织,采用Real-Time PCR检测AURKA基因在不同口腔组织中的表达.结果 AURKA mRNA在OSCC中高表达,且不同分化程度,不同临床分期,组间差异有统计学意义.不同年龄,不同性别,组间差异无统计学意义.结论 AURKA在OSCC中高表达,AURKA作为激酶可能直接或间接地激活多种致癌蛋白或使多种抑癌蛋白失活,从而推动肿瘤的发生发展.     

Polymorphisms of the AURKA (STK15/Aurora Kinase) Gene and Breast Cancer Risk (United States)

AURKA is an important protein in the regulation of G₂ to M transition during mitosis. Due to this regulatory function, it has been hypothesized to be a potential cancer susceptibility gene. Two non-synonymous polymorphisms (F31I and V57I) have been associated with breast cancer risk in prior studies. We sought to confirm these findings in a large case control study nested within a prospective cohort, the Nurses' Health Study. Post-menopausal women who were homozygous for the 31I and 57V alleles had an increased risk of invasive breast cancer (OR 1.63, 95% CI 1.08–2.45). We also performed a meta-analysis to summarize the findings of this and prior studies of association between the F31I polymorphism and breast cancer risk (Summary OR 1.29, 95% CI 1.08–1.53, p-heterogeneity = 0.29). These results confirm prior findings that AURKA represents a low penetrance breast cancer susceptibility gene.     

人AURKA基因RNA干扰慢病毒载体的构建及其对A549细胞增殖的影响

目的:针对AURKA基因构建RNA干扰重组慢病毒质粒并进行慢病毒包装,初步探讨AURKA基因的功能。方法:应用pGCLV-GFP慢病毒载体构建针对AURKA的shRNA载体,转染包装293T细胞,收集病毒上清液,转染肺腺癌细胞株A549。荧光显微镜下观察绿色荧光蛋白表达情况确定转染效率,应用Western blot技术检测AURKA基因在蛋白水平表达的变化,以明确RNAi的抑制率。应用BrdU法分析干扰AURKA前后A549细胞增殖情况的变化,运用克隆形成实验检测干扰AURKA后A549细胞株对长春新碱敏感性的变化。结果:针对AURKA基因的RNAi慢病毒表达载体构建成功,PCR和DNA测序鉴定实验表明插入位点与干扰片断的碱基序列完全正确。在293T细胞中进行慢病毒包装,获得高滴度病毒上清液。通过重组慢病毒载体将AURKA shRNA转导入A549细胞中,转染效率为100%。Western blot检测证实LV-AURKA慢病毒显著抑制AURKA的表达,BrdU检测显示AURKA基因表达下调抑制了A549细胞的增殖。克隆形成实验表明AURKA基因表达增强A549细胞对长春新碱的敏感性。结论:慢病毒载体介导的靶向AURKA的RNA干扰可有效抑制AURKA表达,降低肺腺癌A549细胞的增殖能力,并且增强A549细胞对长春新碱的敏感性。