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1.
Protein & Cell ; (12): 114-122, 2017.
Article in English | WPRIM | ID: wpr-757355

ABSTRACT

Increasing attention is focused on the down-regulation of miRNAs in cancer process. Nuclear receptor subfamily 2 (NR2F2, also known as COUP-TFII) is involved in the development of many types of cancers, but its role in gastric cancer remains elusive. In this experiment, oncomine and Kaplan-meier database revealed that NR2F2 was up-regulated in gastric cancer and that the high NR2F2 expression contributed to poor survival. MicroRNA-27b was targeted and down-regulated by NR2F2 in human gastric cancer tissues and cells. The ectopic expression of miR-27b inhibited gastric cancer cell proliferation and tumor growth in vitro and in vivo. Assays suggested that the overexpression of miR-27b could promote MGC-803 cells' migration and invasion and retard their metastasis to the liver. In addition, down-regulation of miR-27b enhanced GES-1 cells' proliferation and metastasis in vitro. These findings reveal that miR-27b is a tumor suppressor in gastric cancer and a biomarker for improving patients' survival.


Subject(s)
Animals , Female , Humans , Male , Biomarkers, Tumor , Genetics , Metabolism , COUP Transcription Factor II , Genetics , Metabolism , Cell Line, Tumor , Genes, Tumor Suppressor , Heterografts , Mice, Nude , MicroRNAs , Genetics , Metabolism , Neoplasm Metastasis , Neoplasm Proteins , Genetics , Metabolism , Neoplasm Transplantation , RNA, Neoplasm , Genetics , Metabolism , Stomach Neoplasms , Genetics , Metabolism , Pathology
2.
Chinese Medical Sciences Journal ; (4): 157-163, 2004.
Article in English | WPRIM | ID: wpr-253999

ABSTRACT

<p><b>OBJECTIVE</b>To clone and identify the proteins involved in regulating the transcription of hTERT and study the role of genes in both hTERT transcription and telomerase activity.</p><p><b>METHODS</b>The full cDNA of COUP-TFII was cloned from HeLa cDNA library by hTERT promoter-based yeast one-hybrid assay and then in-frame inserted into His-tag fusion expression vector pEK318. The His-tag COUP-TFII fusion proteins were purified by Ni-NTA chromatography. The interaction of COUP-TFII with hTERT promoter in vitro was identified by electrophoretic mobility shift assay and Footprint. The role of COUP-TFII in both hTERT transcription and telomerase activity were probed through Luciferase reporter assay, Northern blot, and TRAP-PCR ELISA.</p><p><b>RESULTS</b>COUP-TFII could firmly bind to the downstream E-box and the other two binding sites in hTERT promoter. Luciferase reporter assay indicated COUP-TFII could suppress hTERT promoter activity and stable introduction of COUP-TFII into HeLa cells also decreased both endogenous hTERT transcription and telomerase activity.</p><p><b>CONCLUSION</b>The human COUP-TFII can firmly bind to hTERT promoter, and inhibit telomerase activity through decreasing hTERT transcription. It will greatly facilitate understanding of telomerase regulation in normal and cancer cells.</p>


Subject(s)
Humans , COUP Transcription Factor II , COUP Transcription Factors , Cloning, Molecular , DNA, Complementary , Genetics , DNA-Binding Proteins , Genetics , Pharmacology , E-Box Elements , Genetics , HeLa Cells , Promoter Regions, Genetic , RNA, Messenger , Genetics , Receptors, Steroid , Genetics , Telomerase , Genetics , Metabolism , Transcription Factors , Genetics , Pharmacology , Transcription, Genetic , Yeasts , Genetics
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