Liu Yan, Wei Jun, Wang Chao, Meng Zihui, Luo Duqiang, Zhao Xiaoling, Hou Ruizhi
Department of Gastrointestinal and Colonretal Surgery, China-Japan Union Hospital of Jilin University, Changchun, China.
Department of Clinical Research, China-Japan Union Hospital of Jilin University, Changchun, China.
J Gastrointest Oncol. 2022 Dec;13(6):3263-3277. doi: 10.21037/jgo-22-1017.
Pancreatic cancer (PC) is one of the most malignant cancers of the gastrointestinal tract. However, the study of targeted therapy research in PC is not very thorough. Therefore, targeted molecular markers are needed to aid in the diagnosis and treatment of PC.
In our research, we investigated the biological functions and molecular mechanism of microRNA-543 in PC. Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to analyze the transcription and protein expression of microRNA-543, Serine/threonine kinase 31 (STK31), and LINC00847 in BxPC-3 and PANC-1 cells. Subsequently, Cell Counting Kit-8 (CCK-8), Transwell, colony formation, and flow cytometry (FCM) assays were utilized to evaluate cell growth, migration, invasion, and apoptosis. WB and fluorescence in-situ hybridization (FISH) were used to evaluate the epithelial-mesenchymal transition (EMT) process and subcellular localization. RNA immunoprecipitation (RIP), double luciferase reporter, and RNA-pull down assays were performed to determine the targeting relationship between microRNA-543 and STK31 or microRNA-543 and LINC00847.
While microRNA-543 expression was discovered to be low in PC, LINC00847 and STK31 were overexpressed at significant levels. MicroRNA-543 knockdown dramatically increased PC cell growth, invasion, metastasis, and EMT, as well as decreased apoptosis in functional studies. Furthermore, microRNA-543 and STK31 were found to be mutual targets. LINC00847 acted as a molecular sponge for microRNA-543 and a competitive endogenous RNA (ceRNA) for STK31, thereby increasing STK-31 transcription.
Our results suggest that microRNA-543, through the LINC00847/microRNA-543/STK31 axis, plays a role in the development of PC as a tumor suppressor. As a result, microRNA-543 may prove to be an effective diagnostic and therapeutic target for PC.
胰腺癌（PC）是胃肠道最恶性的癌症之一。然而，胰腺癌靶向治疗的研究并不十分深入。因此，需要靶向分子标志物来辅助胰腺癌的诊断和治疗。
在我们的研究中，我们研究了微小RNA-543在胰腺癌中的生物学功能和分子机制。采用蛋白质免疫印迹法（WB）和定量实时聚合酶链反应（qRT-PCR）分析微小RNA-543、丝氨酸/苏氨酸激酶31（STK31）和LINC00847在BxPC-3和PANC-1细胞中的转录和蛋白表达。随后，利用细胞计数试剂盒-8（CCK-8）、Transwell小室、集落形成和流式细胞术（FCM）检测评估细胞生长、迁移、侵袭和凋亡。采用WB和荧光原位杂交（FISH）评估上皮-间质转化（EMT）过程和亚细胞定位。进行RNA免疫沉淀（RIP）、双荧光素酶报告基因和RNA下拉实验，以确定微小RNA-543与STK31或微小RNA-543与LINC00847之间的靶向关系。
虽然发现微小RNA-543在胰腺癌中表达较低，但LINC00847和STK31显著过表达。在功能研究中，微小RNA-543敲低显著增加了胰腺癌细胞的生长、侵袭、转移和EMT，同时减少了细胞凋亡。此外，发现微小RNA-543和STK31是相互作用的靶点。LINC00847作为微小RNA-543的分子海绵和STK31的竞争性内源性RNA（ceRNA），从而增加STK-31的转录。
我们的结果表明，微小RNA-543通过LINC00847/微小RNA-543/STK31轴，作为一种肿瘤抑制因子在胰腺癌的发生发展中发挥作用。因此，微小RNA-543可能被证明是胰腺癌有效的诊断和治疗靶点。
