Jin Jinglan, Xu Hongqin, Li Wanyu, Xu Xiaotong, Liu Huan, Wei Feng
Department of Hepatology, The First Hospital of Jilin University, Changchun, China.
Department of Hepatobiliary and Pancreatic Surgery, The First Hospital of Jilin University, Changchun, China.
Front Bioeng Biotechnol. 2020 Feb 18;8:54. doi: 10.3389/fbioe.2020.00054. eCollection 2020.
Hepatocellular carcinoma (HCC) is one of the important types of liver cancer. LncRNA is an important regulatory factor that regulates many biological processes such as tumor cells during tumorigenesis and metastasis. LINC00346 has been associated with various types of liver cancer, but its role and regulatory mechanism in HCC remain unclear. In our study, we found the LINC00356-miR-199a-3p-CDK1/CCNB1 axis through bioinformatics analysis. The expressions of LINC00356, miR-199a-3p, CDK1, and CCNB1 in HCC and normal hepatocytes were determined by qRT-PCR and WB. The results showed that LINC00356, CDK1 and CCNB1 were highly expressed in HCC, while miR-199a-3p was lowly expressed. Dual luciferase reporter gene assay, RIP and RNA-pull down assays demonstrated the targeted binding relationship of LINC00346-miR-199a-3p-CDK1/CCNB1. Overexpressing LINC00460 and silencing miR-199a-3p promoted cell invasion, inhibited apoptosis of HCC, and arrested the cell cycle in S phase while opposite results were obtained when silencing LINC00346, CDK1, and CCNB1. LINC00346 indirectly affects liver cancer by promoting the expression of CDK1/CCNB1 through competitive adsorption of miR-199a-3p. In addition, the study also demonstrated that overexpression of LINC00346 indirectly inhibited the expression of p53 and p21 proteins by promoting CDK1/CCNB1 expressions, thereby blocking the p53 signaling pathway. These results proved that LINC00346 could regulate the expression of CDK1/CCNB1 through the competitive adsorption of miR-199a-3p, thereby affecting the p53 signaling pathway and finally regulating the apoptosis, invasion and cell cycle of HCC cells. In conclusion, LINC00346 can be used as a tumor promoter and potential therapeutic target for HCC metastasis and prognosis.
肝细胞癌（HCC）是肝癌的重要类型之一。长链非编码RNA（LncRNA）是一种重要的调控因子，在肿瘤发生和转移过程中调节许多生物学过程，如肿瘤细胞。LINC00346与多种类型的肝癌有关，但其在HCC中的作用和调控机制仍不清楚。在我们的研究中，通过生物信息学分析发现了LINC00356-miR-199a-3p-CDK1/CCNB1轴。采用qRT-PCR和WB检测HCC和正常肝细胞中LINC00356、miR-199a-3p、CDK1和CCNB1的表达。结果显示，LINC00356、CDK1和CCNB1在HCC中高表达，而miR-199a-3p低表达。双荧光素酶报告基因检测、RNA免疫沉淀（RIP）和RNA下拉实验证实了LINC00346-miR-199a-3p-CDK1/CCNB1的靶向结合关系。过表达LINC00460和沉默miR-199a-3p可促进细胞侵袭，抑制HCC细胞凋亡，并使细胞周期停滞于S期，而沉默LINC00346、CDK1和CCNB1则得到相反的结果。LINC00346通过竞争性吸附miR-199a-3p促进CDK1/CCNB1的表达，从而间接影响肝癌。此外，研究还表明，LINC00346的过表达通过促进CDK1/CCNB1的表达间接抑制p53和p21蛋白的表达，从而阻断p53信号通路。这些结果证明，LINC00346可通过竞争性吸附miR-199a-3p调节CDK1/CCNB1的表达，从而影响p53信号通路，最终调节HCC细胞的凋亡、侵袭和细胞周期。总之，LINC00346可作为HCC转移和预后的肿瘤促进因子和潜在治疗靶点。
