Effect and significance of steroidogenic factor-1 gene silencing on proliferation of human adrenocortical H295R cells
10.3760/cma.j.issn.1000-6702.2010.08.007
- VernacularTitle:类固醇生成因子-1基因沉默对人肾上腺皮质腺癌H295R细胞的作用及其临床意义
- Author:
Dongliang HU
;
Jinzhi OUYANG
;
Xin MA
;
Hongzhao LI
;
Baojun WANG
;
Taoping SHI
;
Shaogang WANG
;
Xu ZHANG
- Publication Type:Journal Article
- Keywords:
Steroidogenic factor-1;
H295R cell;
Adrenal gland neoplasms;
Proliferation
- From:
Chinese Journal of Urology
2010;31(8):524-528
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the influence of inhibited steroidogenic factor-1 on human adrenocortical H295R cells, and explore its role in the pathogenesis of adrenal tumors. Methods The plasmids pGenesil1-SF-1-shRNA which containing U6 promoter and SF-1-specific short hairpin RNA (shRNA) and pGenesil1-negative-shRNA containing unspecific shRNA were transfected into H295R cell. The expression of SF-1 was measured by Western blot and real-time polymerase chain reaction(RT-PCR). Cell proliferation was analyzed by WST-1 assay and cell count. Ki-67 expression was detected by immunohistochemistry and cell apoptosis was examined by TUNEL assay. Results Compared with those in control cells, the protein and mRNA level of SF-1- transfected cells were reduced by 69.7% and 71.2% (P＜0. 01). WST-1 and cell count method showed that SF-1 gene silencing obviously inhibited cell proliferation(P＜0. 01). By contrast, there was a 3. 7-fold increase in the percentage of apoptotic H295R cells in SF-1-inhibited group than that of control group (P＜0. 01). Immunohistochemistry showed that Ki-67 positive cells in SF-1-inhibited cells were lower than the negative control cells (16.90±2.17) % and (33. 48±3.16)%,(P＜0. 01). Conclusion SF-1 gene silencing can inhibit the proliferation of adrenocortical cells, and it is expected to become a key protein in understanding pathogenesis of adrenal tumors or treating them.
