General Surgery, Luhe Hospital, Capital Medical University, Beijing 101100, China.
General Surgery, Tongren Hospital, Capital Medical University, Beijing 101100, China.
Biomed Res Int. 2021 Dec 29;2021:1382061. doi: 10.1155/2021/1382061. eCollection 2021.
To analyze the function of miR-10b-5p in suppressing the invasion and proliferation of primary hepatic carcinoma cells by downregulating erythropoietin-producing hepatocellular receptor A2 (EphA2). . Eighty-six hepatic carcinoma (HCC) tissue specimens and 86 corresponding adjacent tissue specimens were collected, and the mRNA expression of miR-10b-5p and Ephrin type-A receptor 2 (EphA2) in the specimens was determined using a reverse transcription-polymerase chain reaction (RT-PCR) assay. Western blot was employed to quantify EphA2, B-cell chronic lymphocytic leukemia/lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-3 in the cells, and CCK8, Transwell assay, and flow cytometry were applied to evaluate the proliferation, invasion, and apoptosis of cells, respectively. Moreover, the dual luciferase reporter assay was utilized for correlation analysis between miR-10b-5p and EphA2.
miR-10b-5p was lowly expressed in HCC, while EphA2 was highly expressed. Cell experiments revealed that miR-10b-5p overexpression or EphA2 knockdown could reduce cell proliferation, accelerate apoptosis, strongly upregulate Bax and Caspase-3, and downregulate Bcl-2. In contrast, miR-10b-5p knockdown or EphA2 overexpression gave rise to reverse biological phenotypes. Furthermore, dual luciferase reporter assay verified that miR-10b-5p was a target of EphA2, and the rescue experiment implied that transfection of pCMV-EphA2 or Si-EphA2 could reverse EphA2 expression and cell biological functions caused by miR-10b-5p overexpression or knockdown.
miR-10b-5p reduced HCC cell proliferation but accelerate apoptosis by regulating EphA2, suggesting it has the potential to be a clinical target for HCC.
通过下调红细胞生成素产生肝细胞受体 A2（EphA2）分析 miR-10b-5p 抑制原发性肝癌细胞侵袭和增殖的功能。收集 86 例肝癌（HCC）组织标本和 86 例相应的癌旁组织标本，采用逆转录-聚合酶链反应（RT-PCR）检测标本中 miR-10b-5p 和 Ephrin 型-A 受体 2（EphA2）的 mRNA 表达。Western blot 定量检测细胞中 EphA2、B 细胞慢性淋巴细胞白血病/淋巴瘤-2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）和 Caspase-3，CCK8、Transwell 试验和流式细胞术分别用于评估细胞增殖、侵袭和凋亡。此外，还利用双荧光素酶报告基因实验进行 miR-10b-5p 与 EphA2 之间的相关性分析。
miR-10b-5p 在 HCC 中低表达，而 EphA2 高表达。细胞实验表明，miR-10b-5p 过表达或 EphA2 敲低可减少细胞增殖，促进细胞凋亡，强烈上调 Bax 和 Caspase-3，下调 Bcl-2。相反，miR-10b-5p 敲低或 EphA2 过表达则产生相反的生物学表型。此外，双荧光素酶报告基因实验验证了 miR-10b-5p 是 EphA2 的靶基因，挽救实验表明，转染 pCMV-EphA2 或 Si-EphA2 可逆转 miR-10b-5p 过表达或敲低引起的 EphA2 表达和细胞生物学功能。
miR-10b-5p 通过调节 EphA2 减少 HCC 细胞增殖但加速细胞凋亡，提示其具有成为 HCC 临床治疗靶点的潜力。
