Abstract
ObjectiveThe tumor susceptibility gene 101 (TSG101) is closely associated with various tumor types, but its role in the pathogenesis of renal cell carcinoma (RCC) is still unknown. This study used RNA interference to silence the expression of TSG101 in RCC cell lines and explore the role of TSG101 in RCC.MethodsImmunohistochemistry and western blot were performed to detect the expression of TSG101 in 15 paired renal tumor samples. A small interfering RNA (siRNA) targeting TSG101 was transfected into A498 and 786-O cell lines. The Cell Counting Kit-8 (CCK-8) assay and colony formation assay were used to observe the changes in cell proliferation after transfection. Flow cytometry was used to detect the effect on the cell cycle. Western blot was conducted to study the changes of related functional proteins.ResultsThe expression of TSG101 was higher in RCC tissues than in adjacent normal tissues. The CCK-8 assay showed that the proliferation and colony formation of the A498 and 786-O cell lines were attenuated after suppression of TSG101. Flow cytometry showed that silencing of TSG101 induced G0/G1 arrest. The western blot results revealed that the levels of cell cycle-related proteins (c-myc, cyclin E1 and cyclin-dependent kinase 2 (CDK2)) were markedly decreased in the siRNA groups.ConclusionsTSG101 promotes proliferation of RCC cells. This positive effect on tumor growth involves activation of c-myc and cyclin E1/CDK2 and their effect on cell cycle distribution.
Highlights
Renal cell carcinoma (RCC) is one of the most refractory cancers in the world and accounts for about 2 to 3% of adult malignancies [1]
Expression of tumor susceptibility gene 101 (TSG101) was higher in RCC tumor tissues compared with adjacent normal tissues Previous studies have identified TSG101 as an oncogene in various tumor types [19,20,21]
As expected, compared with adjacent normal tissues, TSG101 mRNA expression was higher in RCC tumor tissues (p < 0.05) (Fig. 1b)
Summary
Renal cell carcinoma (RCC) is one of the most refractory cancers in the world and accounts for about 2 to 3% of adult malignancies [1]. TSG101 is a multi-functional protein whose functions include the sorting and transport of endosomes [8,9,10], modulation of protein ubiquitination [11], and participation in p53/MDM2 feedback control loops [12, 13], thereby affecting epithelial cell growth and differentiation [14] and regulation of the cell cycle [15], with significant roles in the maintenance of cell homeostasis. We down-regulated TSG101 using a small interfering RNA (siRNA) in order to observe its impact on the proliferation and cell cycle of RCC cells
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