Abstract
Cancer stem cells (CSCs) are essentially responsible for tumor initiation, growth, progression, metastasis and recurrence, and cigarette smoke (CS) is closely involved in the occurrence and development of kidney cancer. However, the effect of CS on renal CSCs has not been elucidated yet. In the present study, tumorsphere formation assay was used to enrich renal CSCs from 786-O and ACHN cells. We illustrated that CS effectively promoted renal CSCs stemness by enhancing tumorsphere formation, increasing the expression of renal CSCs markers (CD133, CD44, ALDHA1, Oct4, and Nanog) and elevating CD133+ cell population. Moreover, our results showed that CS triggered the activation of Sonic Hedgehog (SHH) pathway, while inhibition of SHH pathway dampened the promotive effects of CS on renal CSCs. Finally, higher levels of renal CSCs markers and SHH pathway-related proteins were observed in kidney cancer tissues from smokers than non-smoking cancer tissues. Taken together, these results demonstrated the important role of SHH pathway in regulating CS-induced renal CSCs stemness augment. Findings from this study could provide new insight into the molecular mechanisms of CS-elicited stemness of renal CSCs.
Highlights
Among urologic tumors, renal cell carcinoma (RCC) is characterized as the highest cancer-specific mortality rate, and the 5-year survival rate for patients with metastatic disease is only 12%1
It was revealed that both protein and mRNA expression levels of the above indicated genes were markedly up-regulated in 786-O and ACHN tumorspheres cultured in serum-free medium (SFM) for 5 days (Figs. 1b, c)
cigarette smoke (CS) promotes the stemness of renal Cancer stem cells (CSCs) To determine the effect of CS extract (CSE) on the viability of the cells, 786-O and ACHN tumorspheres were treated with various concentrations of CSE for 5 days and cell viability was examined by cell counting kit 8 (CCK-8) assay
Summary
Renal cell carcinoma (RCC) is characterized as the highest cancer-specific mortality rate, and the 5-year survival rate for patients with metastatic disease is only 12%1. To evaluate the characteristic of renal CSCs, we cultured two human RCC cell lines 786-O and ACHN under the conditions of SFM and serum-supplied medium (SSM), respectively. It was revealed that both protein and mRNA expression levels of the above indicated genes were markedly up-regulated in 786-O and ACHN tumorspheres cultured in SFM for 5 days
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