Mitotic arrest of gastric cancer cells induced by silencing of STK15 gene.
- Author:
Bin LAN
1
;
Xue-hua CHEN
;
Bing-ya LIU
;
Ying QU
;
Xiao-qing ZHANG
;
Qu CAI
;
Qi-bao DAI
;
Jian ZHANG
;
Zheng-gang ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Adenocarcinoma; metabolism; pathology; Aurora Kinase A; Aurora Kinases; Cell Cycle; drug effects; Cell Line, Tumor; Cell Proliferation; drug effects; DNA, Neoplasm; metabolism; Gene Silencing; Humans; Mitosis; drug effects; Protein-Serine-Threonine Kinases; biosynthesis; genetics; RNA Interference; RNA, Messenger; biosynthesis; genetics; RNA, Small Interfering; pharmacology; Stomach Neoplasms; metabolism; pathology
- From: Chinese Journal of Pathology 2006;35(2):106-109
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the role of STK15 in regulating mitosis of gastric cancer cells (MKN45) by gene silencing through RNA interference mechanism.
METHODSRNA interference technique was used to inhibit STK15 expression in MKN45 cells. The expression levels of STK15 mRNA and protein were measured by real-time quantitative RT-PCR and Western blot respectively and cell morphological changes were investigated by reverse microscopy. In addition, cell cycle distribution and cellular proliferation were determined by flow-cytometry and MTT assay respectively. Finally, the mitotic phenotype of MKN45 cells was studied by immunofluorescence staining and confocal microscopy.
RESULTSSilencing of STK15 gene by RNA interference was confirmed by marked decrease of STK15 mRNA and protein levels in the treated MKN45 cells. This silencing correlated with rounding of the cells, decreasing of DNA content in G(2) phase (P < 0.05) and a lowered proliferation index (P < 0.05), along with alterations of mitotic phenotype of MKN45 (P < 0.05).
CONCLUSIONSTK15 gene may play a key role in regulating cellular mitosis and its inhibition by RNA interference leading to mitosis arrest in MKN45 cells.
