Expression levels of sonic hedgehog pathway genes and their targets are upregulated in early clear cell renal cell carcinoma

Abstract

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 SHH , smoothened ( SMO ), glioma-associated zinc finger protein ( GLI ) 1-3 , BCL2 apoptosis regulator ( BCL2 ), MYC proto-oncogene ( MYC ), vascular endothelial growth factor A ( VEGFA ) and cyclin D1 ( CCND1 ) 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 VEGFA , as well as low CCND1 mRNA expression, were associated with short overall survival, and increased VEGFA 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 2 /M phase and decreased the expression levels of GLI1 , BCL2 , VEGFA and CCND1 . RU-SKI43 inhibited cell migration and decreased the expression levels of BCL2 , MYC and CCND1 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 CCND1 and VEGFA 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 in vivo studies.

Hedgehog Signaling Pathway Orchestrates Human Lung Branching Morphogenesis

“…In CDH, the histological abnormalities observed in the lung are accompanied by lower SHH expression as compared to normal lung development [72]; therefore, we investigated HH pathway expression in CDH (Figures 6 and S4). Consistent with previous reports, we demonstrated a downregulation of the expression of SHH in CDH [57]. Additionally, we show that PTCH1 and HHIP are also decreased in CDH.…”

“…The third class contains inhibitors that act on different phases of production, activation, and binding of the SHH ligand, such as RU-SKI 43 (SHH palmitoylation) or 5E1 (binding) [35,56]. Therefore, we used the SHH antibody 5E1, a strong candidate to safely interfere with HH signaling in clinical trials [57] with limited off-target effects, to abrogate HH activation in our human culture model [17,35]. We showed a strong decrease in the number of branches and an evident increase in cyst dilatation upon HH pathway inhibition, suggesting an involvement in lung branching (Figure 2).…”

bFGF could modulate the anti-cancer effect of Cyclopamine and induce apoptosis in HT-29 colorectal cancer cells

“…Similar to these ndings, our data show that treatment of HT-29 colorectal cancer cell line with cyclopamine at a concentration of 2.5 µmol/L for 48 hours resulted in increased cells apoptosis, increased expression of the Bax and Caspase 3 genes, as well as decreased colony formation. According to another study, inhibiting the Shh pathway using Cyclopamine, can suppress the growth of kidney cancer cells in the G2/M phase of the cell cycle and decrease the expression of the GLI1, BCL2, VEGFA, and Cyclin D1 genes [40]. In contrast, BCL2 and Cyclin D1 gene expression were not decreased in our study by cyclopamine treatment.…”

Resveratrol Induces Apoptosis, Suppresses Migration, and Invasion of Cervical Cancer Cells by Inhibiting the Hedgehog Signaling Pathway

“…Shh was abnormally elevated in various cancers and could serve as a potential biomarker to predict poor prognosis [25]. Downregulation of Shh could effectively inhibit progression of cancer [26][27][28]. It was verified that abnormal activation of Hedgehog pathway could lead to a variety of malignancies, including cervical cancer [29].…”