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S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors.

ABSTRACT: We have previously demonstrated that miR-1236-3p has the robust ability to up-regulate p21 expression by targeting the p21 promoter, thus inhibiting bladder cancer progression. Microarray experiments displayed that miR-1236-3p significantly increased the expression of the oncogenic F-box protein S-phase kinase-associate protein 2 (Skp2) while activating p21 expression in bladder cancer cells. Here, we confirmed that Skp2 was over-expressed following transfection with miR-1236-3p. Further, we demonstrated that miR-1236-3p and its sequence homology dsRNA, dsRNA-245 (which is completely complementary to the p21 promoter), both are able to potently induce p21 expression. We found that dsRNA-245 did not induce changes in Skp2 expression, while miR-1236-3p could increase Skp2 expression; this influence was independent of p21 activation. Moreover, transfection of miR-1236-3p or dsRNA-245 into bladder cancer cells significantly inhibited cell proliferation and clonegenesis and induced cell cycle arrest mainly by regulating p21 expression. However, the growth inhibition caused by dsRNA-245 was more effective than that caused by miR-1236-3p. This difference in effect size is mainly related to the miR-1236-3p-induced expression of Skp2. In summary, our results provided evidence that both endogenous and exogenous small RNAs might function to induce p21 expression by interacting with the same promoter region, therefore impeding bladder cancer cell growth. Additionally, our results indicated that microRNA activation can activate the expression of some tumor suppressor genes as well as some oncogenes. This indicated the need for the further study of clinical applications of RNA activation.

SUBMITTER: Zhang Q

PROVIDER: S-EPMC5883114 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors.

Zhang Qingsong Q Miao Shuo S Li Chuanchang C Cui Kai K Ge Qiangqiang Q Chen Zhong Z

American journal of translational research 20180315 3

We have previously demonstrated that miR-1236-3p has the robust ability to up-regulate p21 expression by targeting the p21 promoter, thus inhibiting bladder cancer progression. Microarray experiments displayed that miR-1236-3p significantly increased the expression of the oncogenic F-box protein S-phase kinase-associate protein 2 (Skp2) while activating p21 expression in bladder cancer cells. Here, we confirmed that Skp2 was over-expressed following transfection with miR-1236-3p. Further, we dem ...[more]

PMID: 29636863

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Project description:Purpose:Lung cancer is one of the most prevailing human cancers worldwide. Emerging evidence implies that long non-coding RNA nicotinamide nucleotide transhydrogenase-antisense RNA1 (NNT-AS1) is implicated in the tumorigenesis of lung cancer. Herein, we aimed to expose the impact of NNT-AS1 on the drug resistance of lung cancer. Methods:Levels of NNT-AS1, microRNA (miR)-1236-3p and autophagy-related gene 7 (ATG7) were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR) assay. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was implemented to detect cell proliferation and the half maximal inhibitory concentration (IC50) of cisplatin (DDP) in vitro. Moreover, flow cytometry was performed to assess cell apoptosis. Cell migration and invasion were examined utilizing transwell assay in lung cancer cells. Besides, levels of ATG7 and cell behavior-related proteins were determined via Western blot. Dual-luciferase reporter assay was administrated to identify the interaction between miR-1236-3p and NNT-AS1 or ATG7. The biological role of NNT-AS1 in DDP ?resistance of lung cancer was examined by xenograft tumor model in vivo. Results:NNT-AS1 and ATG7 were upregulated, whereas miR-1236-3p was curbed in lung cancer tissues and in with or without DDP-resistant cell lines. NNT-AS1 detection significantly constrained cell growth, metastasis, and the IC50 of DDP in A549/DDP and H522/DDP cells. Interestingly, the influence of miR-1236-3p mimic on DDP resistance was overturned via NNT-AS1 upregulation in vitro. Reintroduction of miR-1236-3p inhibitor relieved the effect of ATG7 silencing on DDP sensitivity in A549/DDP and H522/DDP cells. Importantly, NNT-AS1 was a sponge of miR-1236-3p to separate ATG7. Besides, NNT-AS1 silencing enhanced DDP sensitivity of lung cancer in vivo. Conclusion:NNT-AS1/miR-1236-3p/ATG7 axis regulated DDP resistance in lung cancer cells and might supply a probable target and prognostic marker in lung cancer treatment.

Project description:BackgroundAccumulating evidence indicates that circRNAs may serve as essential regulators in the progression of several human cancers, but the function and mechanism of circRNAs in intrahepatic cholangiocarcinoma (ICC) are largely unknown.MethodsRNA-seq was used to assess differentially expressed circRNAs between 4 ICC and peritumor tissues. Quantitative RT-PCR and in situ hybridization were used to determine the circHMGCS1-016 expression in ICC tissues. The function and mechanism of circHMGCS1-016 were further identified via in vivo experiments. The clinical characteristics and prognostic significance of circHMGCS1-016 were analyzed by a retrospective study. The functions of circHMGCS1-016 were assessed via modifying circRNA expression in ICC cells. Moreover, the molecular mechanisms of circHMGCS1-016 in ICC cells were explored by circRNA precipitation, miRNA immunoprecipitation, SILAC and luciferase reporter assays.ResultsWe identified that compared with peritumor tissues, ICC tissues expressed hsa_circ_0008621 (circHMGCS1-016) high by RNA-seq, which was further identified by qRT-PCR and in situ hybridization. Moreover, the expression of circHMGCS1-016 was revealed to be associated with survival and recurrence of ICC patients. By regulating circHMGCS1-016 expression, we found that elevated circHMGCS1-016 promoted ICC development both in vitro and in vivo. By SILAC and circRNA-pull down, we demonstrated that circHMGCS1-016 induced ICC cell invasion and reshaped the tumor immune microenvironment via the miR-1236-3p/CD73 and GAL-8 axis. In ICC tissues, we uncovered that a high level of circHMGCS1-016 was positively associated with CD73 and GAL-8 expression and negatively related to the CD8+ T cells infiltration, which was further validated by establishing a humanized mouse tumor model. Importantly, we displayed that ICC patients with high levels of circHMGCS1-016 in tumor tissues benefited less from anti-PD1 treatment compared to those with low levels of circHMGCS1-016.ConclusionsCircHMGCS1-016 is a forceful contributor in ICC development and immune tolerance via miR-1236-3p/CD73 and GAL-8 axis. CircHMGCS1-016 can be explored as a new potential biomarker and therapeutic target for PD1-resistant ICC.

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S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors.

Publications

S-phase kinase-associated protein 2 impairs the inhibitory effects of miR-1236-3p on bladder tumors.

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