[Effects of RNA interference targeting four different genes on the growth and proliferation of nasopharyngeal carcinoma CNE-2Z cells].
Ying Song, Ming-min Dong, Haifeng Yang
Abstract
Ying Song, Ming-min Dong, Haifeng Yang
Abstract
OBJECTIVE: To explore the effects of RNA interference (RNAi) targeting four different genes (VEGF, c-myc, survivin, hTERT) on the growth and proliferation of nasopharyngeal carcinoma (NPC) CNE-2Z cells. METHODS: Plasmid-1 targeted all four genes, plasmid-2, 3, 4 and 5 targeted VEGF, c-myc, survivin and hTERT respectively. These plasmids were transfected separately into human NPC CNE-2Z cells and xenograft tumors in nude mice. The expressions of plasmids in NPC CNE-2Z cells and xenograft tumors were observed. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western blot respectively. The effects of the plasmids on the biological behavior of CNE-2Z cells were observed with Transwell invasion chamber model. Apoptosis was determined with flow cytometer. The inhibitory effect of the plasmids on xenograft tumors were observed in nude mice. RESULTS: CNE-2Z cell proliferation was significantly inhibited and in vitro invasion ability was significantly decreased in the plasmid-1 group compared with those in the plasmid 2 - 5 groups (all P < 0.05). mRNA and protein expressions of all four genes decreased in the plasmid-1 group. The apoptosis rate in the plasmid-1 group was higher than that in the plasmid 2 - 5 groups (all P < 0.05). Growth of xenograft tumors in nude mice were significantly inhibited in the plasmid 1 - 5 groups, particularly in the plasmid-1 group. CONCLUSION: RNA interference targeting multiple genes can effectively inhibit NPC proliferation and induce apoptosis, which provides an experiment basis for NPC gene therapy.
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OBJECTIVE: To explore the effects of RNA interference (RNAi) targeting four different genes (VEGF, c-myc, survivin, hTERT) on the growth and proliferation of nasopharyngeal carcinoma (NPC) CNE-2Z cells. METHODS: Plasmid-1 targeted all four genes, plasmid-2, 3, 4 and 5 targeted VEGF, c-myc, survivin and hTERT respectively. These plasmids were transfected separately into human NPC CNE-2Z cells and xenograft tumors in nude mice. The expressions of plasmids in NPC CNE-2Z cells and xenograft tumors were observed. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western blot respectively. The effects of the plasmids on the biological behavior of CNE-2Z cells were observed with Transwell invasion chamber model. Apoptosis was determined with flow cytometer. The inhibitory effect of the plasmids on xenograft tumors were observed in nude mice. RESULTS: CNE-2Z cell proliferation was significantly inhibited and in vitro invasion ability was significantly decreased in the plasmid-1 group compared with those in the plasmid 2 - 5 groups (all P < 0.05). mRNA and protein expressions of all four genes decreased in the plasmid-1 group. The apoptosis rate in the plasmid-1 group was higher than that in the plasmid 2 - 5 groups (all P < 0.05). Growth of xenograft tumors in nude mice were significantly inhibited in the plasmid 1 - 5 groups, particularly in the plasmid-1 group. CONCLUSION: RNA interference targeting multiple genes can effectively inhibit NPC proliferation and induce apoptosis, which provides an experiment basis for NPC gene therapy.
Key concepts: Survivin, Nasopharyngeal carcinoma, Plasmid, RNA interference, Cell growth, Transfection, Molecular biology, Telomerase reverse transcriptase