Effect of shRNA interfering PLCε1 gene on proliferation and cell cycle of human esophageal carcinoma Eca 109 cells / 基础医学与临床

ABSTRACT

Objective To explore the impact of silencing PLCε1 gene on proliferation and cell cycle of esophageal carci-noma Eca109 cells.Methods Three plasmid expression vectors (PLCε11, PLCε12 and PLCε13) were constructed to si-lence PLCε1 gene.A negative control plasmid expression vector (HK) was constructed at the same time to serve as a control .The plasmid expression vectors were transfected into esophageal carcinoma Eca 109 cells by cations liposome . The plasmid expression vector with the best interference effect ( PLCε12 ) was chosen .The study included Eca 109 group , HK group and PLCε12 group .Cell viability of Eca 109 cells was evaluated by MTT assay .The cell cycles were detected by FCM .The mRNA expression of P16 and CyclinD1 gene was measured by RT-PCR.Results The cell vi-abilitys of Eca109 cells in PLCε12 group were 80.73%and 75.88%at 48 and 72 h after transfection , which were significantly lower than that of Eca 109 cells in HK group (P<0.001).The percentage of S phase Eca109 cells in PLCε12 group was lower than that of Eca 109 cells in HK group ( P <0.01 ) , the cell cycle of PLCε12 group Eca109 cells was arrested in G0/G1 phase.The P16 gene mRNA expression of PLCε12 group Eca109 cells was higher than that of HK group Eca 109 cells ( P<0.01 ) .Conclusions Silencing PLCε1 gene may up-regulate P16 gene mRNA expression and then arrest the cell cycle at G 0/G1 phase and so inhibit proliferation of Eca 109 cells.