Jiang Wei, Ou Zheng-Lin, Zhu Qin, Yao Yuan-Bing, Zai Hong-Yan
Department of General Surgery, Xiangya Hospital, Central South of University, Changsha 410008, Hunan, PR China.
Department of General Surgery, Xiangya Hospital, Central South of University, Changsha 410008, Hunan, PR China.
Pathol Res Pract. 2022 Apr;232:153829. doi: 10.1016/j.prp.2022.153829. Epub 2022 Feb 25.
Hepatoblastoma is a malignancy that occurs in the liver, most of which occur in children younger than 3 years old. It was reported that lncRNA OIP5-AS1 was up-regulated in hepatoblastoma, but the detailed mechanism by which OIP5-AS1 regulates hepatoblastoma development is unclear.
qRT-PCR, Western blotting, and immunofluorescence were used to examine levels of OIP5-AS1, PTBP1, β-catenin or proliferation/stemness-related molecules. Colony formation, sphere formation, wound healing assay and transwell were applied to detect cell proliferation, stemness and invasion, respectively. RIP assay was used to investigate the interaction of OIP5-AS1/PTBP1 and PTBP1/CTNNB1. Finally, in vivo model was constructed to detect the function of OIP5-AS1 in hepatoblastoma.
OIP5-AS1 was significantly up-regulated in hepatoblastoma cells. OIP5-AS1 silencing notably attenuated the stemness and invasion of hepatoblastoma cells. OIP5-AS1 bound with PTBP1, and silencing of OIP5-AS1 inhibited β-catenin. Meanwhile, overexpression of PTBP1 or β-catenin activation significantly reversed OIP5-AS1 silencing-inhibited hepatoblastoma cell proliferation and stemness. Moreover, β-catenin was found to be the downstream target of PTBP1, and OIP5-AS1 activated β-catenin signaling via promoting the binding between PTBP1 and β-catenin to increase the mRNA stability of β-catenin. Finally, OIP5-AS1 knockdown significantly alleviated the tumor growth of hepatoblastoma by repressing β-catenin.
OIP5-AS1 silencing inhibits the growth and stemness of hepatoblastoma through binding with PTBP1 to inhibit β-catenin signaling pathway. OIP5-AS1 may be the potential target against hepatoblastoma.
肝母细胞瘤是一种发生于肝脏的恶性肿瘤，多数发生于3岁以下儿童。据报道，长链非编码RNA OIP5-AS1在肝母细胞瘤中上调，但其调控肝母细胞瘤发生发展的具体机制尚不清楚。
采用qRT-PCR、蛋白质免疫印迹法及免疫荧光法检测OIP5-AS1、PTBP1、β-连环蛋白或增殖/干性相关分子的水平。分别应用集落形成、成球实验、伤口愈合实验及Transwell实验检测细胞增殖、干性及侵袭能力。采用RNA免疫沉淀实验研究OIP5-AS1/PTBP1及PTBP1/CTNNB1的相互作用。最后，构建体内模型检测OIP5-AS1在肝母细胞瘤中的作用。
OIP5-AS1在肝母细胞瘤细胞中显著上调。沉默OIP5-AS1可显著减弱肝母细胞瘤细胞的干性及侵袭能力。OIP5-AS1与PTBP1结合，沉默OIP5-AS1可抑制β-连环蛋白。同时，PTBP1过表达或β-连环蛋白激活可显著逆转OIP5-AS1沉默对肝母细胞瘤细胞增殖及干性的抑制作用。此外，发现β-连环蛋白是PTBP1的下游靶点，OIP5-AS1通过促进PTBP1与β-连环蛋白的结合激活β-连环蛋白信号通路，增加β-连环蛋白的mRNA稳定性。最后，敲低OIP5-AS1可通过抑制β-连环蛋白显著减轻肝母细胞瘤的肿瘤生长。
沉默OIP5-AS1通过与PTBP1结合抑制β-连环蛋白信号通路，从而抑制肝母细胞瘤的生长及干性。OIP5-AS1可能是肝母细胞瘤的潜在治疗靶点。
