Bax介导硼替佐米诱导胃癌细胞株MKN45凋亡作用的研究

Bortezomib induced apoptosis of MNK45 cells through up-regulation of Bax

目的验证BCl-2相关X(Bax)蛋白在硼替佐米(BTZ)诱导胃癌细胞株MKN45的凋亡作用。方法以人Bax(NC_000019.10)为靶标设计sh RNA序列,将Bax sh RNA转染至MKN45细胞中,G418筛选耐药克隆。荧光定量聚合酶链反应和Western blot测定转染前后Bax m RNA和蛋白含量。给予空载体对照MKN45及sh Bax转染的MKN45细胞BTZ梯度加药(5、25、50、100、250、500和1 000 nmol/L),检测其细胞增殖、凋亡程度。结果通过转染,建立Bax表达稳定下调的MKN45-sh Bax细胞系。给予BTZ梯度加药后,阴性对照(Mock)组胃癌细胞MKN45 Bax含量增加,凋亡程度逐渐增加,增殖率逐渐降低。沉默Bax后BTZ诱导的MKN45细胞凋亡受到抑制,增殖能力增强。结论 Bax能部分介导BTZ对胃癌细胞株MKN45的凋亡作用。

Objective To investigate the effect of Bax on Bortezomib-induced apoptosis of gastric cancer MNK45 cells. Methods Bax shRNA was constructed and transfected into MKN45 ceils. Bortezomib of gradient concentrations was administrated into MKN45 ceils （5 nmol/L, 25 nmol/L, 50 nmol/L, 100 nmol/L, 250 nmol/L, 500 nmol/L and 1,000 nmol/L）. Cell viability and apoptosis rate were examined. Results Bax-knockdown MKN45 cell line was constructed by transfection. Gradient Bortezomib suppressed the proliferation of mock MKN45 cells and induced cell apoptosis, meanwhile Bax protein level was increased. Bortezomib increased apoptosis rate at a concentration of 500 nmol/L. In the shBax group, apoptosis rate was down-regulated and proliferation ability was strengthened. Conclusions Bax-mediated apoptosis caused by Bortezomib and knockdown of Bax could partially attenuate cell death caused by Bortezomib.