Abstract
Purpose To investigate the effects of lncRNA RHPN1-AS1 on retinoblastoma (RB) and further explore its underlying molecular mechanisms. Methods The expression of RHPN1-AS1, miR-3133, (JAK2), and signal transducer and activator of transcription 3 (STAT3) was detected by qRT-PCR. CCK-8, EDU, and flow cytometry assays were conducted to assess the proliferation activity and apoptosis of RB cells. Double fluorescein and RNA immunoprecipitation assays were performed to detect the interaction between RHPN1-AS1 and miR-3133 or miR-3133 and JAK2. Western blotting was performed to detect the expression of apoptosis-related proteins. Results In RB cells, RHPN1-AS1 was upregulated. Silencing RHPN1-AS1 inhibited the activity of RB cells and promoted apoptosis. The expressions of proapoptotic factors (Bax and p53) were increased, while antiapoptotic factors (Bcl-2 and Survivin) were suppressed in siRHPN1-AS1 groups. Furthermore, we predicted and verified that RHPN1-AS1 regulated RB progression by targeting miR-3133/JAK2. In addition, siRHPN1-AS1 also inhibited oncogene STAT3 protein expression. Conclusion lncRNA RHPN1-AS1 served as a sponge for miR-3133 to counteract miR-3133-mediated JAK2/STAT3 suppression, indicating that the lncRNA RHPN1-AS1 may be a potential therapeutic target for the treatment of RB.
Highlights
Retinoblastoma (RB) is a malignant tumor derived from photoreceptor precursor cells that occurs in children, usually under 5 years of age [1]
To further investigate the role of Long noncoding RNAs (lncRNAs) RHPN1-AS1 on RB, Y79 and WERI-Rb1 cells were transfected with siRHPN1-AS1 or siNC and we found that there was a decreased expression of RHPN1-AS1 in the siRHPN1-AS1 group when compared with the siNC group (Figure 1(b))
We performed the Cell Counting Kit-8 (CCK-8) assay, EDU assay, and flow cytometry assay to detect the effects of silencing RHPN1-AS1 on RB cell line proliferation and apoptosis
Summary
Retinoblastoma (RB) is a malignant tumor derived from photoreceptor precursor cells that occurs in children, usually under 5 years of age [1]. There are approximately 8000 newly diagnosed retinoblastoma cases worldwide each year [2]. RB has an excellent prognosis where more than 95% of patients survive their disease and approximately 2 thirds of eyes are salvaged [3]. Most patients with retinoblastoma have a poor prognosis, which is usually due to a lack of multidisciplinary care [4]. RB extends along the optic nerve to the brain and is transferred [5]. Surgery and chemotherapy are still the main antitumor strategies, but chemotherapy has side effects [6]. It is urgent to find useful biomarkers and explore new targets for the treatment of RB
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