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Cripto-1 overexpression is involved in the tumorigenesis of nasopharyngeal carcinoma
BMC Cancer, 09/23/09

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

Results

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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