Abstract:
Objective To investigate the inhibition of Livin expression on colon cancer cell line HT-29 by lentiviral vector-mediated RNA interference (RNAi) and its impact on biological behaviors.
Method Small-interference RNA (siRNA) molecules that target Livin gene were designed, and a pGCSIL-GFP lentiviral vector was created. The inhibitory expression of Livin was detected using Real-time PCR and Western blot. Trypan blue staining was used to detect changes in proliferation. In situ end labeling (TUNEL) and flow cytometry were used to detect changes in apoptosis and cell cycle distribution, respectively. Matrigel invasion assay was used to detect changes in cell invasion ability.
Results Recombinant lentiviral Livin-RNAi-LV1 can efficiently inhibit Livin expression in mRNA and protein level (P < 0.01). Reduced Livin expression can inhibit cell proliferation (P < 0.05), promote cell apoptosis (P < 0.01), and re-distribute the cell cycle, leading to increased proportion of G0/G1 phase cells (P < 0.05) and reduced S phase cells (P < 0.05) and G2/M phase cells < 0.05). In vitro invasive ability was significantly damaged (P < 0.01).
Conclusion The inhibition of Livin expression on HT-29 cell by Lentiviral vector-mediated RNA interference can inhibit proliferation, promote apoptosis, and reduce cell invasiveness; thus, it deserves further study.