Department of Histology, Faculty of Medicine, Medical University of Gdansk, 80211 Gdansk, Poland.
Department of Urology, Faculty of Medicine, Medical University of Gdansk, 80402 Gdansk, Poland.
Int J Mol Med. 2022 May;49(5). doi: 10.3892/ijmm.2022.5114. Epub 2022 Mar 10.
Clear cell renal cell carcinoma (ccRCC) is the most common and aggressive subtype of kidney cancer, with high mortality rates worldwide. The sonic hedgehog (SHH) molecular cascade is altered in various malignancies in tumorigenesis, and several SHH pathway inhibitors have been considered as potential anticancer drugs. The aim of the present study was to determine the expression profile of SHH signaling components and their target genes in ccRCC. Additionally, the present study examined the effects of SHH pathway inhibitory drugs (RU‑SKI43, cyclopamine and GLI‑antagonist 61) on cell viability, cell cycle progression, expression levels of SHH target genes and migration ability in 786‑O, ACHN and HK2 cells. The study also included paired tumor and normal samples from 62 patients with ccRCC. The mRNA levels in clinical samples and cell lines were measured via reverse transcription‑quantitative PCR. Cell viability was examined using a sulforhodamine B assay. Flow cytometry was used to investigate cell cycle progression and the migratory rate of cells was assessed using a wound healing assay. High mRNA levels of , smoothened (), glioma‑associated zinc finger protein (), BCL2 apoptosis regulator (), MYC proto‑oncogene (), vascular endothelial growth factor A () and cyclin D1 () were observed in the tumor tissues, especially in early ccRCC, according to the TNM stage or World Health Organization/International Society of Urological Pathology (ISUP) grade. High expression levels of , as well as low mRNA expression, were associated with short overall survival, and increased expression was an independent prognostic factor of a poor outcome in patients with advanced ISUP grade (Cox hazard ratio test). Cyclopamine treatment was found to arrest 786‑O cells in the G/M phase and decreased the expression levels of , , and . RU‑SKI43 inhibited cell migration and decreased the expression levels of , and in ACHN cells. Overall, the results of the present study suggested that SHH signaling may be involved in the early development of ccRCC, and the expression levels of and may serve as prognostic factors of this disease. Cyclopamine and RU‑SKI43 appear to be potential anti‑renal cell carcinoma drugs; however, this hypothesis requires verification by further studies.
肾透明细胞癌 (ccRCC) 是最常见且侵袭性最强的肾癌亚型，全球死亡率较高。 sonic hedgehog (SHH) 分子级联在肿瘤发生的各种恶性肿瘤中发生改变，几种 SHH 途径抑制剂已被认为是潜在的抗癌药物。本研究旨在确定 SHH 信号传导成分及其靶基因在 ccRCC 中的表达谱。此外，本研究还研究了 SHH 途径抑制药物 (RU-SKI43、环巴胺和 GLI-拮抗剂 61) 对 786-O、ACHN 和 HK2 细胞活力、细胞周期进程、SHH 靶基因表达水平和迁移能力的影响。该研究还包括来自 62 名 ccRCC 患者的配对肿瘤和正常样本。通过逆转录-定量 PCR 测量临床样本和细胞系中的 mRNA 水平。使用磺酰罗丹明 B 测定法检测细胞活力。通过流式细胞术研究细胞周期进程，通过划痕愈合试验评估细胞迁移率。根据 TNM 分期或世界卫生组织/国际泌尿病理学会 (ISUP) 分级，在肿瘤组织中观察到高 mRNA 水平的 、 smoothened ()、glioma-associated zinc finger protein ()、BCL2 apoptosis regulator ()、MYC proto-oncogene ()、vascular endothelial growth factor A () 和 cyclin D1 ()，尤其是在早期 ccRCC 中。高水平的 表达与总生存期短有关，而低 mRNA 表达与晚期 ISUP 分级患者的不良预后相关（Cox 风险比检验）。环巴胺处理导致 786-O 细胞停滞在 G/M 期，并降低 、 、 和 的表达水平。RU-SKI43 抑制 ACHN 细胞迁移并降低 、 和 的表达水平。总体而言，本研究结果表明 SHH 信号可能参与 ccRCC 的早期发展，而 表达水平可能作为该疾病的预后因素。环巴胺和 RU-SKI43 似乎是潜在的抗肾细胞癌药物；然而，这一假设需要进一步研究来验证。
