[Mitotic arrest of gastric cancer cells induced by silencing of STK15 gene].
OBJECTIVE
To investigate the role of STK15 in regulating mitosis of gastric cancer cells (MKN45) by gene silencing through RNA interference mechanism.


METHODS
RNA 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.


RESULTS
Silencing 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).


CONCLUSIONS
STK15 gene may play a key role in regulating cellular mitosis and its inhibition by RNA interference leading to mitosis arrest in MKN45 cells.
Lan
Bin
B
Department of Surgery, Shanghai Institute of Digestive Surgery, Ruijin Affiliated Hospital of Shanghai Second Medical University, Shanghai 200025, China.
Chen
Xue-hua
XH
Liu
Bing-ya
BY
Qu
Ying
Y
Zhang
Xiao-qing
XQ
Cai
Qu
Q
Dai
Qi-bao
QB
Zhang
Jian
J
Zhu
Zheng-gang
ZG
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology; D064096:Aurora Kinase A; D064127:Aurora Kinases; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004273:DNA, Neoplasm / Q000378:metabolism; D020868:Gene Silencing; D006801:Humans; D008938:Mitosis / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis* / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology*; D013274:Stomach Neoplasms / Q000378:metabolism* / Q000473:pathology
D004273:DNA, Neoplasm; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C576917:AURKA protein, human; D064096:Aurora Kinase A; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases