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Cripto-1 overexpression is involved in the tumorigenesis of nasopharyngeal carcinoma
BMC Cancer, 09/18/09
Wu Z et al. – Cripto-1 overexpression is connected with the tumorigenesis and progression of NPC, lentivector-mediated RNAi might be feasible for the inhibition of the growth and invasion of NPC.
Methods- The expression of Cripto-1 at mRNA level was detected by the reverse transcription-polymerase chain reaction (RT-PCR) and real time RT-PCR, and western blot was used to examine the protein expression
- Cripto-1 expression and its clinical characteristics were investigated by performing immunohistochemical analysis on a total of 37 NPC clinical tissue samples
- Lentiviral vectors were constructed to get an efficient expression of anti-Cripto-1 siRNA in CNE-2 and C666-1 cells, with invalid RNAi sequence as control
- After the inhibition of the endogenous Cripto-1, the growth, cell cycle and invasion of cells were detected by MTT, FACS and Boyden chamber assay respectively.
- In vivo, the proliferation of the tumor cells was evaluated in xenotransplant nude mice model with whole-body visualizing instrument
- The results of real-time RT-PCR and western blot showed that the expression level of Cripto-1 was markedly higher in NPC cell lines than that in the immortalized nasopharyngeal epithelial cell at both mRNA and protein levels
- RT-PCR of 17 NPC tissues showed a high expression rate in 76.5% (13/17) cases
- In an immunohistochemical study, Cripto-1 was found to express in 54.1% (20/37) cases of NPC
- In addition, Cripto-1 overexpression was significantly associated with N classification (p=0.034), distant metastasis (p=0.036), and clinical stage (p=0.007)
- Inhibition of endogenous Cripto-1 by lentivirus-mediated RNAi silencing technique suppressed NPC cell growth and invasion in vitro
- In vivo, the average weight (p=0.026) and volume (p=0.044) of tumor in CNE-2/GFP+/Cripto-1- xenotransplant mice group were significantly lower than those in the control group
- The Ki67 index was obviously lower in Cripto-1 RNAi treated tumors
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