摘要
目的观察丝氨酸/苏氨酸激酶15基因(STK15)沉默后对胃癌细胞MKN45凋亡的诱导作用,阐述STK15基因对胃癌细胞生存的关键作用。方法化学合成小片断干扰RNA(siRNA)抑制MKN45细胞STK15基因的表达；实时定量PCR及Western印迹检测STK15 mRNA及蛋白质表达的变化；流式细胞仪检测MKN45周期分布及凋亡的变化；Hoechest染色观察凋亡形态变化,Western印迹检测pro-caspase 3水平的变化。结果经靶向STK15的siRNA作用48 h后,STK15 siRNA mRNA及蛋白质表达明显下降；较多MKN45细胞呈现G_2期细胞DNA含量(P＜0．05)；细胞凋亡率较对照组明显上升(P＜0．05)；伴随pro-caspase 3水平下降。结论抑制STK15基因表达通过caspase 3途径导致MKN45细胞凋亡,STK15基因对胃癌细胞增殖及存活起着关键作用。
Objective To observe the effect of tinhibition of serine/threonine kinasel5 （STK15） gene expression on apoptosis induction in gastric cancer cell line-MKN45 and discuss the role of STK15 in viability of gastric cancer cells. Methods The STK15 expression was inhibited by chemically synthesized siRNA. The STK15 mRNA and protein level were respectively measured by real-time quantitative PCR and western blotting, the change of cell cycle distribution and apoptosls rate were detected by flow-cytometry, cell morphological change was observed by Hoechest staining, and pro-caspase 3 level was also detected by western blot. Results After treatment by siRNA targeting STK15 after 48 h, STK15 mRNA and protein level decreased obviously. More MKN45 cells accumulated at G2/M phase （P 〈 0. 05）. The apoptosis rate of STK15 siRNA treated MKN45 cells was higher than that of control cells（ P 〈 0. 05） with the pro-caspase 3 level decreased. Conclusions Inhibition of STK15 gene expression may induce apoptosis in MKN45 cells through the pathway of caspase3. STK15 gene play a key role in proliferation and viability of MKN45 cells.
出处
《中华胃肠外科杂志》
CAS
2006年第5期417-420,共4页
Chinese Journal of Gastrointestinal Surgery
基金
国家973重点基础研究发展规划基金(2002CB713700)
福建医科大学科学研究发展基金(FJGXY04027)
