Long non-coding RNA colon cancer-associated transcript 1-Vimentin axis promoting the migration and invasion of HeLa cells / 中华医学杂志(英文版)

BACKGROUND@#Long non-coding RNA colon cancer-associated transcript 1 (CCAT1) is involved in transforming multiple cancers into malignant cancer types. Previous studies underlining the mechanisms of the functions of CCAT1 primarily focused on its decoy for miRNAs (micro RNAs). However, the regulatory mechanism of CCAT1-protein interaction associated with tumor metastasis is still largely unknown. The present study aimed to identify proteome-wide CCAT1 partners and explored the CCAT1-protein interaction mediated tumor metastasis.@*METHODS@#CCAT1-proteins complexes were purified and identified using RNA antisense purification coupled with the mass spectrometry (RAP-MS) method. The database for annotation, visualization, and integrated discovery and database for eukaryotic RNA binding proteins (EuRBPDB) websites were used to bioinformatic analyzing CCAT1 binding proteins. RNA pull-down and RNA immunoprecipitation were used to validate CCAT1-Vimentin interaction. Transwell assay was used to evaluate the migration and invasion abilities of HeLa cells.@*RESULTS@#RAP-MS method worked well by culturing cells with nucleoside analog 4-thiouridine, and cross-linking was performed using 365 nm wavelength ultraviolet. There were 631 proteins identified, out of which about 60% were RNA binding proteins recorded by the EuRBPDB database. Vimentin was one of the CCAT1 binding proteins and participated in the tumor metastasis pathway. Knocked down vimetin ( VIM ) and rescued the downregulation by overexpressing CCAT1 demonstrated that CCAT1 could enhance tumor migration and invasion abilities by stabilizing Vimentin protein.@*CONCLUSION@#CCAT1 may bind with and stabilize Vimentin protein, thus enhancing cancer cell migration and invasion abilities.

Effect of miR-200c Regulation of Peptidyl-Prolyl Cis/Trans Isomerase on the Biological Behavior of Hep-2 Cells / 中国医科大学学报

Objective To explore the influence of miR-200 c on the biological behavior of laryngeal carcinoma Hep-2 cells and determine whether miR-200 c exerts its biological function through peptidyl-prolyl cis/trans isomerase (PIN1) in laryngeal carcinoma. Methods A qRT-PCR assay for the expression of miR-200 c was performed in laryngeal carcinoma tissues. Hep-2 cells were transfected with miR-200 c related small RNAs. Transwell assay detected the migration ability of the cells. Immunofluorescence assay was used to detect the abnormal amplification of the centrosome. A dual luciferase reporter gene system was used to detect the binding ability between miR-200 c and PIN1. Western blotting detected the protein expression level of PIN1. Results The expression of miR-200 c in laryngeal carcinoma was significantly increased. miR-200 c inhibited the migration of Hep-2 cells and could weaken the abnormal amplification of centrosome.PIN1 was confirmed as one of the target genes of miR-200 c. miR-200 c inhibited the expression of PIN1 at the translation level and could inhibit Hep-2 cell migration and abnormal centrosome amplification by regulating PIN1. Conclusion miR-200 c can inhibit the migration ability of laryngeal carcinoma cells and abnormal centrosome amplification by regulating PIN1.