Effects of rapamycin on expression of ten kinds autophagy-related miRNAs in RAW264.7 macrophages
10.3969/j.issn.1000-484X.2014.08.011
- VernacularTitle:雷帕霉素对RAW264.7巨噬细胞10种与细胞自噬相关的miRNAs表达的影响
- Author:
Tao ZHANG
;
Ximing ZHENG
;
Linlin ZHOU
;
Xin LIU
;
Jin ZHAO
;
Guangxian XU
- Publication Type:Journal Article
- Keywords:
miRNA;
Rapamycin;
real-time PCR
- From:
Chinese Journal of Immunology
2014;(8):1055-1058,1063
- CountryChina
- Language:Chinese
-
Abstract:
To detect the influence of rapamycin on the expression of miR-30b,miR-200a and miR-17-5p etc in macrophages and provide the basis to study the regulation of miRNA in autophagy mechanism of macrophages .Methods: Small RNA was extracted at different times after stimulated with rapamycin in cultured RAW 264.7 cells.After using the stem-loop reverse transcription primers to reverse transcribed into cDNA ,the expression of miR-30b ,miR-30c,miR-106a,miR-214,miR-183,miR-200a, miR-376c,miR-17-5p, miR-142-3p, miR-377 was detected by Real-Time PCR.Results: After RAW264.7 cells was treated by rapamycin for 2,4,6 and 8 hours,the expression of miR-17-5p and miR-106 increased (More than 2.1 times,P<0.05) in 2,4 and 6 hours;miR-214 was up regulated in 2 and 8 hours (More than 2.4 times,P<0.05);miR-30b,miR-30c,miR-183,miR-200a,miR-376c and miR-142-3p was up regulated in 2,6 and 8 hours (2.4 times,P<0.05 );while miR-183 and miR-200a was down regulated at 4 hours(More than 2.1 times,P<0.05);miR-30b was significantly low expression in 8 hours (more than 50 times,P<0.05);miR-377 was up regulated at 4 hours (more than 2.5 times,P<0.05),but was significantly down regulated at 2 and 8 hours (More than 50 times,P<0.05) Conclusion: The expression of miR-200a,miR-30b,miR-377,miR-30c,miR-376c and miR-17-5p is significantly changed after rapamycin stimulated RAW264.7 macrophages,indicated the miRNA may plays an important role in autophagy through the regulation of autophagy-related genes.
