Inhibitory effect of nasopharyngeal carcinoma CNE-2 cells through targeted inhibitory effect on the Bcl-2 gene expression by Hsa-mir-15a/16-1

OBJECTIVE To observe the inhibitory effect of Targeted inhibition of Bcl-2 gene expression by Hsa-miR-15a/16-1 on the growth of nasopharyngeal carcinoma CNE-2 cells.METHODS The recombinant plasmid pENTR-CMV-EGFP-hsa-mir-15a/16-1 which had been successfully constructed was transfected into CNE-2 cells with the help of Lipofectamine 2000.The expression of miR-15a/16-1,Bcl-2 mRNA was detected by RT-qPCR.The levels of Bcl-2 and Caspase-3 Protein were measured by Western blot and FCM.The inhibitory effect of Bcl-2 gene on growth of CNE-2 cells was detected by CCK-8.Apoptosis of the CNE-2 cells was observed by analyzing of nucleus morphology with DAPI pigmentation.RESULTS After transfection of the recombinant plasmid,the expression of both miR15a and miR-16-1 increased significantly(F=547.525,P 0.001;F =99.979,P 0.001),the expression of Bcl-2 mRNA had no significant difference(F=0.220,P 0.05),the expression of Bcl-2 protein decreased,the expression of activated Caspase-3 protein increased.Apoptosis of CNE-2 cell in vitro was obvious.Meanwhile the cell proliferation was inhibited after transfection of the recombinant plasmid,and the effects showed an obvious time-effect relationship.CONCLUSION Targeted inhibition of Bcl-2 gene expression by Hsa-miR-15a/16-1 could inhibit cells proliferation and promote apoptosis on human nasopharyngeal carcinoma CNE-2 cells in vitro.