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[Effect of lentivirus-mediated NOB1 gene silencing by RNA interference on proliferation and apoptosis of human colon cancer cells].

OBJECTIVE
To investigate the effect of NOB1 gene on proliferation and apoptosis of human colon cancer cell line RKO by RNA interference.


METHODS
Small interference RNA(siRNA) targeting NOB1 gene was cloned into lentivirus vector. Then the lentivirus particles expressing NOB1 short harpin RNA(shRNA) were infected into RKO cells. Real-time PCR and Western blot were performed to examine the expression of NOB1 in lentivirus infected cells. The Thermo Scientific Cellomics ArrayScan VTI HCS Reader was used to test the proliferation and colony-formation of RKO cells, and flow cytometry assay was performed to detect cell cycle and apoptosis. Xenograft tumor was established by injection of RKO cells into nude mice, then NOB1-shRNA was injected into the tumor and tumor volume was detected.


RESULTS
Compared to negative controls, the expression levels of NOB1 mRNA and protein were both significantly down-regulated, the proliferation and colony-forming capacity of RKO cells were significantly inhibited, and cell apoptosis was increased after 3 days of NOB1-shRNA lentivirus infection(all P<0.05). The tumor volume was significantly smaller in NOB1-shRNA group than that in Scr-shRNA group[(405±102) mm(3) vs.(870±165) mm(3), P<0.05].


CONCLUSION
Silencing NOB1 gene by RNA interference may provide an inhibitive effect on human colon cancer development.