Inhibitory effect and its molecular mechanisms of MicroRNA-34a on human na-sopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice
10.3969/j.issn.1000-484X.2015.07.011
- VernacularTitle:MicroRNA-34 a对人鼻咽癌CNE-2细胞裸鼠皮下移植瘤的生长抑制作用及分子机制研究
- Author:
Haoxuan ZHANG
;
Yin JIA
- Publication Type:Journal Article
- Keywords:
MicroRNA-34a;
Human nasopharyngeal carcinoma;
CNE-2 cell line;
Nude mice;
Subcutaneous xenograft tumor;
Inhibitory effect;
Molecular mechanisms
- From:
Chinese Journal of Immunology
2015;(7):912-916
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the inhibitory effect and its possible molecular mechanisms of MicroRNA-34a(miR-34a) on the human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice.Methods: The human nasopharyngeal carcinoma CNE-2 cell line was cultured in vitro.miR-34a and Scrambled miRNA recombinant plasmids were successfully established and stably transfected into CNE-2 cells.Fifteen six-week-old male nude mice were divided randomly into three groups:miR-34a group(5 mice) ,Scrambled miRNA group(5 mice) ,Blank control group(5 mice).Different CNE-2 cells were subcuta-neously injected on the back near right lower limb.Tumor volumes were examined every 7 days.Mice were executed on the 35 days,and the eventual average tumor volumes and weights were examined.Total RNA and protein were isolated from tumors,and the expression of miR-34a,CDK6,and Bcl-2 mRNA and protein were determined by qRT-PCR and western blot,respectively.Results: The relative expressions of miR-34a was significantly up-regulated in miR-34a transfected group compared to Scrambled miRNA transfected group (P<0.05).Eight subcutaneous xenograft tumor models were successfully established.The nude mice of miR-34a transfected group appeared a smaller tumor volume compared to the other two groups at the beginning of 21th day( P<0.05).The eventual average tumor volumes in miR-34a group,Scrambled miRNA group and blank control group were(351.37±98.19)mm3,(798.75±91.04)mm3 and
(849.62±101.32) mm3 ,respectively,and the eventual average tumor weights in miR-34a group,Scrambled miRNA group and blank control group were(0.81±0.13)g,(1.47±0.21)g and(1.58±0.37)g,respectively.Both the eventual average tumor volumes and weights in miR-34a group were lower compared to the other two groups(P<0.05).qRT-PCR results revealed that the expression of miR-34a in miR-34a transfected group was significantly higher than in the other two groups,while the mRNA and protein expression of CDK6 and Bcl-2 were lower than the other two groups ( P<0.05 ) .Conclusion: miR-34a may inhibit the growth of human nasopharyngeal carcinoma CNE-2 cell line subcutaneous xenograft tumor in nude mice by down-regulating CDK6 and Bcl-2.
