Abstract

This study aimed to explore the effects of RNAi-mediated cathepsin L gene silencing on bionomics of hepatoma carcinoma cells. In this study, cathepsin L siRNA silencing group (silencing group), blank hepatoma carcinoma cells group (blank group) and the siRNA fluorescin group (fluorescence control group) were set. The observing time include 1, 3 and 6 days after RNA interference for cathepsin L. The transfection efficiency of each group was observed. The expression of cathepsin L in hepatoma carcinoma cells was detected by immunofluorescence, reverse transcription polymerase chain reaction (RT-PCR) and western blot. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay. The changes of cell cycle and apoptosis were observed by flow cytometry. The changes of invasiveness of hepatoma carcinoma cells were detected by Boyden chamber. Compared with the blank group and fluorescence control group, mRNA and protein level of cathepsin L decreased significantly, and cell growth was inhibited. Meanwhile, the proliferation index decreased significantly, while the apoptosis index increased significantly in the experimental group. The invasiveness of hepatoma carcinoma cells was also found to decrease. The study indicated that RNA interference could inhibit cathepsin L expression, and decrease cell proliferation and cell invasiveness of hepatoma carcinoma cells efficiently.    

Key words: RNA interference, liver cancer; Cathepsin L. 

Abbreviation

RT-PCR, Reverse transcription polymerase chain reaction; MTT,methyl thiazolyl tetrazolium; RNAi, RNA interference; DMEM, Dulbecco’s modified Eagle’s medium; GFP, green fluorescent protein; DMSO, dimethyl sulfoxide; PBS,phosphate buffer; PI, propidium Iodide; FCS, fecal calf serum; PCI, protein C inhibitor; FCM, flow cytometric.