The different expressed microRNA in adrenocorticotropin-independent macronodular adrenal hyperplasia / 中华泌尿外科杂志

ABSTRACT

Objective To investigate the different expressed microRNA in adrenocorticotropinindependent macronodular adrenal hyperplasia (AIMAH).Methods For the screening different expressed microRNA in AIMAH,5 AIMAH fresh tissues and 5 normal adrenal fresh tissues were collected.Total RNA was extracted by using Trizol reagents,then was purified and tested by denaturing agarose gel electrophoresis and NanoDrop 2000.Then the purified RNA was labeled by using Hy3TM fluorescent label and hybridized with miRCURY Array.MicroRNA array was scanned and analyzed by GenePix 4000B microarray scanner and GenePix proV6.0.The ratio acquired by divided AIMAH fluorescent correction value by normal adrenal fluorescent correction value.If the ratio was greater than 3.0,the microRNA was defined as up-regulating,or smaller than 0.3,as down-regulating.For the validation 40 AIMAH samples and 10 normal adrenal samples were used.The miRCURY array result was validated by real-time quantitative PCR.All the data was managed by SPSS 17.0.When P ＜ 0.05,the difference was significant in Student's t test.Results There were 12 different expressed microRNA between normal adrenal gland and AIMAH,including 7 up-regulating and 5 down-regulating microRNAs.Up-regulating microRNAs included hsa-miR-663,hsa-miR-498,hsa-miR-638,hsa-miR-501-5p,hsa-miR-585,hsa-miR-557 and hsa-miR-144.Down-regulating microRNA included hsa-miR-744,hsa-miR-143,hsa-miR-26a,hsa-miR-22,hsa-miR-29a.Four different expressed microRNAs were identified in validation test,including 3 up-regulating and 1 down-regulating microRNA.Upregulating microRNA included has-miR-663,has-miR-498,and has-miR-557.Down-regulating microRNA was has-miR-744.Conclusion Compared with normal adrenal gland,there might be some different expressed microRNAs in AIMAH.