Abstract:

Objective To investigate the expression of histone demethyltransferases JARID1B in bladder cancer tissues and human bladder cancer cell line T24, construct JARID1B lentiviral expression vector and perform identification, and investigate the expression of JARID1B infected by packed lentivirus. Methods The expression of JARID1B protein in 6 cases of bladder cancer tissues, tissues adjacent to cancer, and human bladder cancer cell line T24 was detected by Western blotting. With eukaryotic expression plasmid pcDNA3.1（-）-JARID1B, JARID1B gene sequence was incorporated into lentiviral vectors pLv-enhanced green fluorescent protein (EGFP) which contained EGFP, and recombinant lentivirus expression vector pLv-EGFP-JARID1B was constructed. After DNA sequence analysis, pLv-JARID1B plasmid was transfected into T24 by Lipofectamine, the expression of GFP was observed by fluorescence microscopy, and the expression of JARID1B mRNA was detected by RT-PCR. Lentivirus was packed and T24 was infected, and the expression of JARID1B protein wes determined by Western blotting. Results The positive expression rates of JARID1B protein in bladder cancer tissues and tissues adjacent to cancer were 16.7% and 66.7%, respectively, and the expression was weaker for the former. The expression of JARID1B protein in T24 was negative. JARID1B sequence was verified to be inserted into lentiviral vectors pLv-EGFP. After transfection of T24 with pLv-EGFP-JARID1B, the expression of GFP was observed by fluorescence microscope, and the expression of JARID1B mRNA in T24 was detected by RT-PCR. After infection of T24 by packed lentivirus, Western blotting analysis revealed that the expression of JARID1B protein was detected.ConclusionThe expression of JARID1B is down-regulated in bladder cancer tissues. JARID1B lentiviral expression plasmid can be successfully constructed, and there is overexpression of JARID1B in T24 infected with packed lentivirus.

Key words: histone demethylation, JARID1B, lentivirus, gene expression, bladder cancer