Zhang Li-Yun, Liu Xin, Zheng Shuo, Xi Wen-Qun, Wu Xue-Ping, Nie Dan-Yao, Hu Hui-Ling, Wang Jian-Tao, Liu Xin-Hua, Zhang Jing
Shenzhen Eye Hospital; Jinan University; Shenzhen Eye Institute, Shenzhen 518000, Guangdong Province, China.
Int J Ophthalmol. 2024 Aug 18;17(8):1387-1395. doi: 10.18240/ijo.2024.08.01. eCollection 2024.
To investigate the impact of hsa_circ_0007482 on the proliferation and apoptosis of human pterygium fibroblasts (HPFs) and its correlation with the severity grades of pterygium.
Pterygium and normal conjunctival tissues were collected from the superior area of the same patient's eye (=33). The correlation between pterygium severity and hsa_circ_0007482 expression using quantitative reverse-transcription polymerase chain reaction (RT-qPCR) were analyzed. Three distinct siRNA sequences targeting hsa_circ_0007482, along with a negative control sequence, were transfected into HPFs. Cell proliferation was assessed using the cell counting kit-8. Expression levels of Ki67, proliferating cell nuclear antigen (PCNA), Cyclin D1, Bax, B-cell lymphoma-2 (Bcl-2), and Caspase-3 were measured RT-qPCR. Immunofluorescence staining was employed to detect Ki67 and vimentin expressions. Apoptosis was evaluated using flow cytometry.
Hsa_circ_0007482 expression was significantly higher in pterygium tissues compared to normal conjunctival tissues (<0.001). Positive correlations were observed between hsa_circ_0007482 expression and pterygium severity, thickness, and vascular density. Knockdown of hsa_circ_0007482 inhibited cell proliferation, reducing the mRNA expression of Ki67, PCNA, and Cyclin D1 in HPFs. Hsa_circ_0007482 knockdown induced apoptosis, increasing mRNA expression levels of Bax and Caspase-3, while decreasing Bcl-2 expression in HPFs. Additionally, hsa_circ_0007482 knockdown attenuated vimentin expression in HPFs.
The downregulation of hsa_circ_0007482 effectively hampers cell proliferation and triggers apoptosis in HPFs. There are discernible positive correlations detected between the expression of hsa_circ_0007482 and the severity of pterygium.
探讨hsa_circ_0007482对人翼状胬肉成纤维细胞(HPFs)增殖和凋亡的影响及其与翼状胬肉严重程度分级的相关性。
从同一患者眼睛的上方区域收集翼状胬肉组织和正常结膜组织(n = 33)。使用定量逆转录聚合酶链反应(RT-qPCR)分析翼状胬肉严重程度与hsa_circ_0007482表达之间的相关性。将三种靶向hsa_circ_0007482的不同小干扰RNA序列以及一个阴性对照序列转染到HPFs中。使用细胞计数试剂盒-8评估细胞增殖。通过RT-qPCR测量Ki67、增殖细胞核抗原(PCNA)、细胞周期蛋白D1、Bax、B细胞淋巴瘤-2(Bcl-2)和半胱天冬酶-3的表达水平。采用免疫荧光染色检测Ki67和波形蛋白的表达。使用流式细胞术评估细胞凋亡。
与正常结膜组织相比,翼状胬肉组织中hsa_circ_0007482的表达显著更高(P < 0.001)。观察到hsa_circ_0007482表达与翼状胬肉严重程度、厚度和血管密度之间呈正相关。敲低hsa_circ_0007482可抑制细胞增殖,降低HPFs中Ki67、PCNA和细胞周期蛋白D1的mRNA表达。敲低hsa_circ_0007482可诱导细胞凋亡,增加HPFs中Bax和半胱天冬酶-3的mRNA表达水平,同时降低Bcl-2表达。此外,敲低hsa_circ_0007482可减弱HPFs中波形蛋白的表达。
hsa_circ_0007482的下调有效抑制了HPFs的细胞增殖并触发细胞凋亡。hsa_circ_0007482的表达与翼状胬肉的严重程度之间存在明显的正相关。