Wuchang Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, Hubei, China.
Bioengineered. 2021 Dec;12(1):6144-6154. doi: 10.1080/21655979.2021.1972785.
Abnormally high expression of RecQ protein-like 4 (RECQL4) has been observed in many cancers, including hepatocellular carcinoma (HCC). We aimed to explore the effects of RECQL4 on HCC progression and the possible mechanisms. RECQL4 expression in HCC tissues and its correlation with the prognosis of HCC patients were analyzed using GEPIA2 and UALCAN databases. After detecting RECQL4 levels in several human HC cell lines, RECQL4 was silenced by siRNA transfection. Cell viability, migration and invasion were tested with CCK-8, wound healing and transwell assays. The levels of epithelial-mesenchymal transition (EMT) proteins were evaluated by western blotting. The ENCORI database was adopted for the analysis of the correlation between RECQL4 and serine/arginine splicing factor 1 (SRSF1) in HCC tissues. RNA immunoprecipitation and actinomycin D addition assay were employed to evaluate the combination of these two genes. SRSF1 was overexpressed to assess the biological function of HCC cells with RECQL4 silencing. Results suggested that RECQL4 was overexpressed in HCC tissues and cell lines, which was related to poor prognosis of HCC patients. RECQL4 loss-of-function repressed the proliferation, migration, invasion and EMT of HCC cells. RECQL4 was positively correlated with SRSF1 in HCC tissues. Moreover, SRSF1 was confirmed as an RNA binding protein of RECQL4. Further experiments found that SRSF1 knockdown reduced the stability of RECQL4 mRNA. Rescue assays indicated that SRSF1 overexpression crippled the braking effects of RECQL4 knockdown on the progression of HCC cells. Collectively, SRSF1 can bind to RECQL4 mRNA and enhance its stability, thereby promoting the progression of HCC.
RecQ 蛋白样 4（RECQL4）的异常高表达已在许多癌症中观察到，包括肝细胞癌（HCC）。我们旨在探讨 RECQL4 对 HCC 进展的影响及其可能的机制。使用 GEPIA2 和 UALCAN 数据库分析 HCC 组织中 RECQL4 的表达及其与 HCC 患者预后的相关性。在检测几种人 HCC 细胞系中的 RECQL4 水平后，通过 siRNA 转染沉默 RECQL4。使用 CCK-8、划痕愈合和 Transwell 测定法测试细胞活力、迁移和侵袭。通过 Western blot 评估上皮-间充质转化（EMT）蛋白的水平。采用 ENCORI 数据库分析 HCC 组织中 RECQL4 与丝氨酸/精氨酸剪接因子 1（SRSF1）的相关性。采用 RNA 免疫沉淀和放线菌素 D 加药试验评估这两个基因的结合。过表达 SRSF1 以评估沉默 RECQL4 后 HCC 细胞的生物学功能。结果表明，RECQL4 在 HCC 组织和细胞系中高表达，与 HCC 患者的不良预后相关。RECQL4 功能丧失抑制 HCC 细胞的增殖、迁移、侵袭和 EMT。RECQL4 在 HCC 组织中与 SRSF1 呈正相关。此外，SRSF1 被确认为 RECQL4 的 RNA 结合蛋白。进一步的实验发现，SRSF1 敲低减少了 RECQL4 mRNA 的稳定性。挽救实验表明，SRSF1 过表达削弱了 RECQL4 敲低对 HCC 细胞进展的制动作用。总之，SRSF1 可以与 RECQL4 mRNA 结合并增强其稳定性，从而促进 HCC 的进展。
