The Ministry of Education Key Laboratory of Laboratory Medical Diagnostics, the College of Laboratory Medicine, Chongqing Medical University, Yuzhong District, Chongqing, 400016, China.
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, 650223, China.
In Vitro Cell Dev Biol Anim. 2021 May;57(5):501-509. doi: 10.1007/s11626-021-00589-z. Epub 2021 May 17.
Metastasis is inevitable in about 30% of patients with primary renal cell carcinoma after nephrectomy treatment. APOBEC1 complementation factor (A1CF), an RNA binding protein, participates in tumor progressions such as growth, apoptosis, differentiation, and invasion. Here, we explored biological functions of A1CF and provided a new insight into renal cell carcinoma metastasis. Wound healing assay was conducted to detect migration in A1CF overexpression and knockdown stable cell lines. Quantitative PCR and western blot assays were utilized to test transcriptional and translation levels of A1CF and SMAD3 in A1CF overexpression and knockdown renal carcinoma cells. Nuclear and cytoplasmic protein separation assays were conducted to evaluate the subcellular distribution of A1CF and SMAD3. Immunoprecipitation assay was conducted to detect the interaction between A1CF and SMAD3. Our study demonstrated A1CF overexpression facilitated cell migration in renal carcinoma cells. A1CF deficiency downregulated expression of SMAD3, Snail1, and N-cadherin. In addition, A1CF promoted nucleus translocation of SMAD3 and interacted with SMAD3. SMAD3 knockdown attenuated cell migration induced by A1CF overexpression. Our study suggested A1CF facilitated cell migration by promoting nucleus translocation of SMAD3 in renal cell carcinoma cells.
转移是大约 30%接受肾切除术治疗的原发性肾细胞癌患者不可避免的。APOBEC1 补体因子 (A1CF)，一种 RNA 结合蛋白，参与肿瘤的生长、凋亡、分化和侵袭等进展。在这里，我们研究了 A1CF 的生物学功能，并为肾细胞癌转移提供了新的见解。通过划痕愈合实验检测 A1CF 过表达和敲低稳定细胞系中的迁移。利用定量 PCR 和 Western blot 检测 A1CF 过表达和敲低肾癌细胞中转录和翻译水平的 A1CF 和 SMAD3。通过核质蛋白分离实验评估 A1CF 和 SMAD3 的亚细胞分布。通过免疫沉淀实验检测 A1CF 和 SMAD3 之间的相互作用。我们的研究表明 A1CF 过表达促进肾癌细胞的迁移。A1CF 缺失下调 SMAD3、Snail1 和 N-钙粘蛋白的表达。此外，A1CF 促进 SMAD3 的核转位并与 SMAD3 相互作用。SMAD3 敲低减弱了 A1CF 过表达诱导的细胞迁移。我们的研究表明，A1CF 通过促进肾癌细胞中 SMAD3 的核转位促进细胞迁移。
