Effects Of Agkihpin, An Arginine Esterase From Venom, In Nasopharyngeal Carcinoma CNE-2 Cell Line

Objective:To explore the effects of Agkihpin on the expressions of multidrug resistance associated protein 1, Cyclooxygenase-2 and E-Cadherin in nasopharyngeal carcinoma CNE-2 cell line, and try to describe the mechanism of Agkihpin inhibiting CNE-2.Methods:The cultured cells were treated with different concentrations of Agkihpin for 72 h. The expression level of MRP1, COX-2 and E-CD in CNE-2 cell line was assayed by RT-PCR, immunocytochemistry and Western blotting.Results:1. Comparing with Agkihpin absent group (control group), the MRP1 expression levels in CNE-2 cells were reduced significantly after the treatment of different concentrations of Agkihpin (4.13,0.516 and 0.207μg/ml) (P<0.01), in a concentration-dependent manner, which show that Agkihpin could down-regulate the expressions of MRP1 in CNE-2 cells significantly. After being treated with Agkihpin at different concentrations, the transcription of MRP1 mRNA in CNE-2 cells decreased significantly as compared with control group(P<0.01). Immunocytochemistry analysis revealed a decrease in expression of MRP1 protein in these groups (P<0.01), and the down-regulation rate of the three mRNA in CNE-2 was:60.53%,45.21% and 26.95%, respectively. Western blotting showed the similar results on the protein levels expression by immunocytochemisrty.2. Comparing with Agkihpin absent group, the COX-2 levels expression in CNE-2 cells were reduced significantly after the treatment of different concentrations of Agkihpin (4.13,0.516 and 0.207μg/ml) (P<0.01), in a concentration-dependent manner, which show that Agkihpin could down-regulate the expressions of COX-2 in CNE-2 cells significantly. After being treated with Agkihpin at different concentrations, the transcription of COX-2 mRNA in CNE-2 cells decreased significantly as compared with control group(P<0.01). Immunocytochemistry analysis revealed a decrease in expression of COX-2 protein in these groups (P<0.01), and the down-regulation rate of the three mRNA in CNE-2 was:52.17%,32.82% and 26.74%, respectively. Western blotting showed the similar results on the protein expression levels by immunocytochemisrty.3. Comparing with Agkihpin absent group, the E-CD expression levels in CNE-2 cells were reduced significantly after the treatment of different concentrations of Agkihpin (4.13,0.516 and 0.207μg/ml) (P<0.01), in a concentration-dependent manner, which show that Agkihpin could down-regulate the expressions of E-CD in CNE-2 cells significantly. After being treated with Agkihpin at different concentrations, the transcription of E-CD mRNA in CNE-2 cells decreased significantly as compared with control group(P<0.01). Immunocytochemistry analysis revealed a decrease in expression of E-CD protein in these groups (P<0.01), and the down-regulation rate of the three mRNA in CNE-2 was:36.66%,31.28% and 14.21%, respectively. Western blotting showed the similar results on the protein expression levels by immunocytochemisrty.Conclusion:1. Agkihpin can inhibit the expression of MRP 1 in CNE-2 cells, which be likely to the one of reasons for Agkihpin inhibiting the cellular vitality of nasopharyngeal carcinoma cell, and be likely to enhance the sensitivity of CNE-2 cells to chemotherapeutic drugs to some extent.2. Agkihpin can inhibit the expression of COX-2 in CNE-2 cells, which be likely to the one of reasons for Agkihpin inhibiting the cellular vitality of nasopharyngeal carcinoma cell; Agkihpin may be useful in therapy of nasopharyngeal carcinoma.3. Agkihpin can inhibit the expression of E-CD in CNE-2 cells, which would be likely to the one of reasons for Agkihpin inhibiting the cellular vitality of nasopharyngeal carcinoma cell.4. Agkihpin would be likely to inhibit the cellular vitality of CNE-2 cells with down-regulating MRP1, while it would be likely to inhibit the proliferation and migration with down-regulating the COX-2 and E-CD.