Objective   To construct a shRNA expression plasmid targeting phospholipase C epsilon (PLCε) and identify its function in bladder cancer cell line T24.    Methods   Potential RNAi oligonucleotides of PLCε were selected with the aid of online program. Then these oligonucleotides were synthesized and pGenesil-PLCε plasmids were constructed using pGenesil vector containing green fluorescent protein (GFP). The plasmids were digested by

Sal

Ⅰ and sequenced to confirm the inserted sequence. RT-PCR and Western blot analysis were taken to detect the expression of PLCε at RNA and protein levels. The func

tion of this PLCε RNAi in T24 cells was analyzed by MTT assay and flow cytometry.    Results   It was confirmed by digesting and sequencing that the 2 recombinant plasmids had been constructed successfully. The result of RT-PCR and Western blotting showed that pGenesil-PLCε plasmids inhibited PLCε expression in T24 cells obviously

and flow cytometry showed that the cell cycle was arrested at G

0

/G

1

phase. MTT assay showed that proliferation of T24 cells was obviously inhibited.    Conclusion   pGenesil-PLCε effectively inhibits PLCε expression and cell proliferation in the bladder cancer cells.