Kalantzakos Thomas J, Sebel Luke E, Trussler James, Sullivan Travis B, Burks Eric J, Sarita-Reyes Carmen D, Canes David, Moinzadeh Alireza, Rieger-Christ Kimberly M
Department of Translational Research, Lahey Hospital & Medical Center, Burlington, MA 01805, USA.
Department of Urology, Lahey Hospital & Medical Center, Burlington, MA 01805, USA.
Biomedicines. 2022 Sep 28;10(10):2425. doi: 10.3390/biomedicines10102425.
Differential microRNA (miRNA) expression can portend clear cell renal cell carcinoma (ccRCC) progression. In a previous study, we identified a subset of dysregulated miRNA in small renal masses, pT1 ccRCC (≤5 cm) that are associated with an aggressive phenotype. The present study investigated miRNA expression in clinical stage I (cT1) tumors (≤5 cm), comparing pathologic stage I (pT1) tumors to those upstaged to pathologic stage 3 (pT3) after surgery following identification of renal vein invasion or invasion into adjacent fat tissue within Gerota's fascia. Twenty cT1 tumors were examined in an miRNA screening, 10 pT1 and 10 pT3 tumors. The ccRCC cell lines 786-O and Caki-1 were used to assess the impact of let-7c-5p and its protein target insulin-like growth factor 1 receptor (IGF1R). Cells were transfected with pre-let-7c-5p and assessed through cell proliferation, migration, and invasion assays. IGF1R expression was evaluated through Simple Western, and interaction between let-7c-5p and IGF1R was confirmed via luciferase reporter assay. Screening identified 20 miRNA, including let-7c-5p, that were dysregulated between pT1 and pT3 upstaged tumors. This miRNA was also downregulated in our previous study of pT1 tumors that progressed to metastatic disease. Transfection of ccRCC cells with pre-let-7c-5p significantly inhibited proliferation, migration, invasion, and IGF1R expression. These findings suggest that miRNA dysregulation is involved in ccRCC progression, specifically through invasion, and that let-7c-5p downregulation contributes to the aggressiveness of small ccRCC tumors, in part, through its regulation of IGF1R.
差异性微小RNA(miRNA)表达可预示透明细胞肾细胞癌(ccRCC)进展。在先前的一项研究中,我们在小肾肿块、pT1期ccRCC(≤5 cm)中鉴定出一组失调的miRNA,其与侵袭性表型相关。本研究调查了临床I期(cT1)肿瘤(≤5 cm)中的miRNA表达,将病理I期(pT1)肿瘤与在术中发现肾静脉侵犯或肾周筋膜内邻近脂肪组织侵犯后病理分期上调至病理3期(pT3)的肿瘤进行比较。在miRNA筛查中检测了20个cT1肿瘤、10个pT1肿瘤和10个pT3肿瘤。使用ccRCC细胞系786 - O和Caki - 1评估let - 7c - 5p及其蛋白靶点胰岛素样生长因子1受体(IGF1R)的影响。用pre - let - 7c - 5p转染细胞,并通过细胞增殖、迁移和侵袭试验进行评估。通过Simple Western评估IGF1R表达,并通过荧光素酶报告基因试验证实let - 7c - 5p与IGF1R之间的相互作用。筛查鉴定出20种miRNA,包括let - 7c - 5p,在pT1和上调至pT3的肿瘤之间失调。在我们先前对进展为转移性疾病的pT1肿瘤的研究中,这种miRNA也下调。用pre - let - 7c - 5p转染ccRCC细胞可显著抑制增殖、迁移、侵袭和IGF1R表达。这些发现表明,miRNA失调参与ccRCC进展,特别是通过侵袭,并且let - 7c - 5p下调部分通过其对IGF1R的调节促成小ccRCC肿瘤的侵袭性。