- In the current study, we aimed to investigate the clinical impact and roles of TRIM27 in the development of RCC.. - Methods: The mRNA levels of TRIM27 and Kaplan – Meier survival of RCC were analyzed from The Cancer Genome Atlas database. - Real-time PCR and Western blotting were used to measure the mRNA and protein levels of TRIM27 both in vivo and in vitro. - Results: We discovered that TRIM27 was elevated in RCC patients, and the expression of TRIM27 was closely correlated with poor prognosis. - TRIM27 directly interacted with I κ b α , an inhibitor of NF- κ B, to promote its ubiquitination, and the inhibitory effects of TRIM27 on I κ b α led to NF- κ B activation.. - TRIM27 serves as a specific prognostic indicator for RCC, and strategies targeting the suppression of TRIM27 function may shed light on future therapeutic approaches.. - To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.. - The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.. - However, the role of TRIM27 in RCC has not yet been investigated.. - The gene expression of TRIM27 and Kaplan–Meier ana- lysis were downloaded from TCGA dataset (http://. - ualcan.path.uab.edu/analysis.html) to analyze the effect of TRIM27 on overall survival of RCC patient. - Knockdown and overexpression of TRIM27. - All animal procedures were performed according to the Institutional Animal Care and Use Committee. - siNC- or siTRIM27-transfected Caki-2 cells (n = 2 × 10 6. - Roche, Switzerland) prin- cipally according to the supplier’s instructions. - Increased expression of TRIM27 in RCC correlates with poor prognosis. - Western blots showed that TRIM27 was in- creased in 786–0 and Caki-1 cells at translational levels (Fig. - Kaplan–Meier analysis of TCGA datasets demon- strated that the survival time of patients with high mRNA levels of TRIM27 was significantly shorter than that of patients with lower levels (Fig. - suggest that up-regulation of TRIM27 expression in RCC links to a shorter survival duration. - To gain a better understanding of the clinical signifi- cance of TRIM27, we studied the biological actions of TRIM27 using RCC cell lines, and by altering TRIM27 expression in vivo.. - Suppression of TRIM27 inhibits cell proliferation and promotes apoptosis. - We designed three small interference RNAs (siRNAs) targeting TRIM27 and examined the effects on Caki-2 and 786–0 cells with a relatively higher level of TRIM27.. - The mRNA level of TRIM27 was upregulated in renal cancer. - Prognosis of RCC (KIRC) patients with high or low expression of TRIM27 derived from the TCGA database. - The relative mRNA level of TRIM27 was upregulated in human renal cancer tissues. - TRIM27 siRNAs deeply suppressed the relative mRNA and protein content of TRIM27 in human Caki-2 and 786 – 0 cells, respectively. - The mRNA levels of TRIM27 was used 2 −ΔΔCt method and normalized to that of GAPDH. - siTRIM27 – 1 and siTRIM27 – 2 significantly suppressed the proliferation of Caki-2 and 786 – 0 cells. - Apoptosis of Caki-2 and 786 – 0 cells was significantly increased following transfecting with siTRIM27 – 1 and siTRIM27 – 2 at 48 h, respectively. - All three siRNAs effectively inhibited the mRNA and protein expression of TRIM27 (Fig. - We used CCK-8 to determine the effects of TRIM27 on cell proliferation and found that cell growth was dramat- ically suppressed in TRIM27-suppressed groups in both Caki-2 (Fig. - 2C) and 786–0 cells (Fig. - We next used Annexin V-FITC/PI staining to evaluate the effects of TRIM27 on apoptosisand found that sup- pression of TRIM27 in RCCs promoted cell apoptosis (Fig. - The role of NF-κB translocation to the nucleus in inhib- ition of apoptosis is well defined [10]. - Our results demonstrated that NF-κB was less active in the presence of TRIM27 siR- NAs (Fig. - Moreover, our results suggested that knockdown of TRIM27 promoted the expression of IkBa in Caki-2 and 786–0 cells respectively.. - OeTRIM27 rescue the function of TRIM27 in siTRIM27- transfected cells. - To further verify our results, oeTRIM27 was used to res- cue the function of TRIM27 in siTRIM27-transfected cells. - Furthermore, the protein level of TRIM27 was significantly recovered by oeTRIM27 in siTRIM27- transfeced cells. - Taken together, these results suggest that the function of TRIM27 is rescued by oeTRIM27 in siTRIM27- transfected cells.. - Suppression of TRIM27 reduces tumor growth in vivo Next, we examined whether TRIM27 siRNAs reduce tumor development in vivo. - Our results illustrated that apoptosis was increased in the siTRIM27 group compared to the control group. - These findings suggest that suppression of TRIM27 prevents tumor growth in vivo, partially due to increased tumor cell apoptosis (Fig. - To further determine the function of TRIM27 in vivo, the oeNC or oeTRIM27 transfected Caki-1 cells were used to construct a xenograft model. - TUNEL staining was performed to examine apoptosis in oeNC and oeTRIM27 tumors, and we found that overexpression of TRIM27 significantly suppressed the apoptosis of Caki-1 cells in vivo (Fig. - 2F), we sought to determine whether NF- κB mediates the actions of TRIM27. - Importantly, the in- hibitor PDTC also suppressed the protein level of TRIM27 in oeTRIM27 transfected cells, while pro- moted the expression of IkBa.. - We found there was a linear correlation between levels of TRIM27 and NF-κB (Fig. - These data suggest that NF-κB is a potential upstream regulator of TRIM27 in RCC. - Iκbα has been identified as the major regulator of the NF-κB pathway [19], and dissociation of NF-κB from Iκbα is a critical step in translocation to the nucleus. - 3 oeTRIM27 rescues the function of TRIM27 in siTRIM27-transfected cells. - The proliferations of Caki-2 and 786 – 0 cells transfected with siTRIM27, oeTRIM27, or siTRIM27 + oeTRIM27 at and 48 h respectively. - Flow cytometric evaluation of the apoptosis of Caki-2 and 786 – 0 cells transfected with siTRIM27, oeTRIM27, or siTRIM27 + oeTRIM27 at 48 h respectively. - Western blot was used to examine the relative proteins of TRIM27, cleaved caspase-3, NF- κ B (cytoplasm), and NF- κ B (nuclear) in Caki-2 and 786 – 0 cells transfected with siTRIM27, oeTRIM27, or siTRIM27 + oeTRIM27 at 48 h respectively.. - potential uses of TRIM27 as a candidate target for effective molecular-targeted therapeutics for RCC.. - Knockdown of TRIM27 in Caki-2 cells deeply suppressed the tumor volume and weight in vivo. - Overexpression of TRIM27 in Caki-2 cells significantly increased the tumor volume and weight in vivo. - The distinctly different actions of TRIM27 as com- pared to other TRIM superfamily members are likely due to the diversity of protein structures. - 6 The NF- κ B inhibitor PDTC suppressed the function of oeTRIM27 in Caki-1 cells. - oeTRIM27 significantly upregulated the relative mRNA and protein levels of TRIM27 in Caki-1 cells. - However, similar to the molecular biological features of TRIM27 in esophageal and colon cancer [5, 7], we identified similar biological functions of TRIM27 in that it is significantly elevated in. - Many downstream signaling pathways of TRIM27 have been identified in cancer, including PTEN/AKT, p38, and STAT . - Western blot was used to examine the protein content of TRIM27 and I κ b α in oeNC or oeTRIM27 with or without the MG132 treatment. - TRIM27 interacted with I κ b α in human Caki-2 cells. - Further investigation is required to clarify the actions of TRIM27 through other pathways in RCC.. - The online version contains supplementary material available at https://doi.. - org/10.1186/s . - The relative mRNA levels of TRIM27 were much higher in human renal 786 – 0 and Caki-2 cells than that of HK-2 cells. - Western blot was performed to examine the protein contents of TRIM27 in HK A498, ACHN, CaKi-1 and CaKi-2 cells respectively. - S2: siNF- κ B inhibited the expression of TRIM27 in Caki-1 cells. - The relative mRNA and protein levels of NF- κ B and TRIM27 in Caki-1 cells that transfecting with siNC, siNF- κ B-1, siNF- κ B-2 and siNF- κ B-3 respectively. - agreed to submit to the current journal. - All animal procedures were performed according to the Institutional Animal Care and Use Committee.. - https://doi.org/10.1200/EDBK_279905.. - https://doi.org/10.1016/j.. - https://doi.org/10.18388/pb.2019_250.. - https://doi.org . - https://doi.org/10.3892/ijo . - https://doi.org/10.1146/a nnurev-virology . - 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