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Sheng Gao,1 Qingxia Chu,1 Xia Liu,1 Xia Zhao,2 Libao Qin,1 Guoliang Li,1 Qinghuai Liu3 1Department of Ophthalmology, Nanjing Pukou Central Hospital, Nanjing 211800, People’s Republic of China; 2Department of Ophthalmology, Tangshan Eye Hospital, Tangshan 063000, People’s Republic of China; 3Department of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, People’s Republic of ChinaCorrespondence: Qinghuai LiuDepartment of Ophthalmology, The First Affiliated Hospital of Nanjing Medical University, 300# Guangzhou Road, Gulou District, Nanjing 210029, People’s Republic of ChinaEmail liuqinghuai6304@hotmail.comBackground: Abnormally expressed long noncoding RNA (lncRNA) high expression in hepatocellular carcinoma (HEIH) has been implicated in many types of human cancer, and plays crucial roles in tumor development and progression. However, little is known about its function in retinoblastoma.Methods: qRT-PCR was used to determine the expression levels of HEIH, miR-194-5p and WEE1 in retinoblastoma tissues and cell lines. The trypan blue exclusion method, colony formation assay, wound-healing assay and transwell invasion assay were performed to evaluate the effects of HEIH, miR-194-5p and WEE1 on cell proliferation, migration and invasion. Bioinformatics analysis, dual-luciferase reporter assay and Western blot were employed to investigate the regulatory relationship among HEIH, miR-194-5p and WEE1.Results: We found that HEIH was up-regulated in retinoblastoma tissues and cell lines. Furthermore, high level of HEIH was associated with TNM stage, optic nerve invasion and choroidal invasion of patients with retinoblastoma. Functional studies showed that HEIH knockdown significantly suppressed retinoblastoma cell proliferation, migration and invasion. Mechanistically, HEIH promoted retinoblastoma progression by serving as a sponge of miR-194-5p to regulate WEE1 expression.Conclusion: Our work suggests that HEIH acts as an oncogenic lncRNA to promote retinoblastoma proliferation and metastasis, providing a new insight into the retinoblastoma treatment.Keywords: retinoblastoma, HEIH, miR-194-5p, WEE1, metastasis