Inhibitory effect of BRAFV600E gene silencing on proliferation of papillary thyroid cancer cells / 肿瘤

ABSTRACT

Objective: To investigate the effect of V600E mutation of v-raf murine sarcoma viral oncogene homolog B1 (BRAFV600E) gene silencing by RNA interference on the proliferation of papillary thyroid cancer cells. Methods: The plasmid vectors containing short hairpin RNA (shRNA) targeting BRAFV600E gene (BRAFV600E-shRNA) were transfected into papillary thyroid cancer CGTHW3 cells. The silencing efficiency of BRAFV600E-shRNA was measured by semi-quantitative reverse transcription-PCR (RT-PCR) and Western blotting, and the related proteins in mitogen extracellular kinase/extracellular signal-regulated kinase (MEK/ERK) signal pathway were detected by Western blotting. Then the proliferation and cell cycle distribution of CGTHW3 cells after BRAFV600EshRNA transfection were detected by MTT method and flow cytometry, respectively. Results: The expression levels of BRAFV600E mRNA and protein were significantly reduced in CGTHW3 cells transfected with specific shRNAs, and the expressions of Ras, phospho-MEK (p-MEK) and p-ERK proteins in MEK/ERK signal pathway were depressed (all P V600E-shRNA was significantly inhibited (P V600E-shRNA transfection group was higher than that in the blank control group, while the percentage of cells at S phase was reduced (both P V600E gene silencing can inhibit the proliferation of papillary thyroid cancer cells through blocking MEK/ERK signal pathway.