A Molecular Mechanism Of N,N'-dinitrosopiperazine-induced AGR2 Gene Expression To Participate In The Invasion And Metastasis Of Nasopharyngeal Carcinoma

Objective: In the present, the invasion and metastasis of nasopharyngeal carcinoma(NPC) cell with N,N'-dinitrosopiperazine(DNP) treatment and its anterior gradient 2(AGR2),Cathepsin B(CTSB) and Cathepsin D(CTSD) gene expressions were detected to investigate the mechanisms of DNP-regulated AGR2,CTSB and CTSD expression participated in NPC metastasis.Methods: 1. The NPC cell line with low metastasis 6-10 B and NPC cell line with high metastasis 5-8F were served as cells models. 6-10 B cells were treated with DNP(100?M), the cells with treated with 0.1% dimethyl sulfoxide(DMSO) served as the control, and their invasion and metastatic abilities were detected. 2. Wound healing assay were used to detect the motility of 6-10 B cells with DNP treatment. 3. After being treated with DNP, the motility and invasion of 6-10 B cells were detected using Transwell assay. 4. Real-time reverse transcription polymerase chain reaction(RT-PCR) was used to detect AGR2, CTSB and CTSD gene expressions in 6-10 B cells with DNP treatment and 5-8F. 5. The data were analyzed by software SPSS13.0.Results: 1. Wound healing assay on the motility of 6-10 B cells with DNP treatment showed that the invaded cells dramatically increased in 6-10 B cell with DNP treatment when compared with the blank control. 2. Transwell assay on the invasion and metastasis of 6-10 B cells with DNP treatment showed that the invaded cells dramatically increased in 6-10 B cell with DNP treatment when compared with the blank control. DNP could increase the motility and invasion of 6-10 B cells. 3. Real-time RT-PCR data showed that AGR2, CTSB and CTSD m RNA expression rate were 1.54 ± 0.06(P<0.05), 0.80 ± 0.07(P>0.05) and 0.84 ± 0.10(P>0.05) in the DNP treated 6-10 B cells. AGR2, CTSB and CTSD m RNA expression rate were 1.93±0.52(P<0.05), 1.39±0.25(P>0.05), 1.32±0.11(P>0.05) in the 5-8F cells. AGR2 gene expression was significantly higher in the treated cells than that in the control.Conclusions: AGR2 gene expressions are associated with NPC metastasis. DNP-mediated AGR2 gene expression may be a main factor for NPC metastasis. DNP treatment could not increase CTSB and CTSD gene expressions.