Effect of silencing Kinesin family member 21B expression in hepatoma cells BEL-7404 on cell biological behavior

Abstract

Objective To detect the differential expression of Kinesin family member 21B (KIF21B) gene in hepatocellular carcinoma (HCC) tissues and normal tissues and its influence on cell. Methods The Cancer Genome Atlas (TCGA) database was used to screen the differential expression of KIF21B gene, the expression level of KIF21B gene in hepatocellular carcinoma cell lines was detected by reverse transcriptase-polymerase chain reaction (RT-PCR), and the short hairpin RNA (shRNA) targeting KIF21B gene was constructed to infect BEL-7404 cells. The expression of KIF21B mRNA in transfected hepatoma cells was detected by real-time quantitative polymerase chain reaction (Real-time PCR), and the expression of KIF21B protein was detected by Western blotting. The effect of KIF21B gene depletion on cell proliferation was detected by Celigo. KIF21B gene subtraction was detected by methyl thiazol tetrazolium (MTT) assay. The effect on cell proliferation was detected by flow cytometric fluorescence sorting (FACS). The effect of KIF21B gene depletion on apoptosis was detected by colony formation assay. The effect of KIF21B gene subtraction on cell clone formation ability was detected by colony formation assay. The measurement data were expressed as mean±standard deviation (Mean±SD), and t-test was used for comparison between groups. Results Compared with normal tissues, the expression of KIF21B gene was up-regulated in 31 HCC tissues, unchanged in 14 and down-regulated in 5. The expression level of KIF21B in 4 HCC cell lines was higher than that in normal Liver tissue cells (P<0.05). ShRNA targeting KIF21B effectively silenced the expression of KIF21B, and MTT assay showed that the growth rate of bel-7404 cells was significantly inhibited after KIF21B reduction (P<0.05). Apoptosis was significantly increased (P<0.05). The clonal formation ability of KIF21B cells was significantly decreased (25.3±3.7 vs. 418.2±26.5, P<0.05). Conclusion After KIF21B gene silencing, the apoptosis of hepatocellular carcinoma cells increased, the proliferation and cloning ability decreased significantly. Key words: Hepatoma cells; Kinesin family member 21B; Apoptosis; Proliferation