Bax介导硼替佐米诱导胃癌细胞株MKN45凋亡作用的研究

发布时间：2018-10-31 07:39

【摘要】：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。
[Abstract]:Objective to investigate the effect of BCl-2 associated X (Bax) protein on the apoptosis of gastric cancer cell line MKN45 induced by bortezomie (BTZ). Methods sh RNA sequence was designed with human Bax (NC_000019.10) as target and Bax sh RNA was transfected into MKN45 cells. Drug resistant clones were screened by G418. Fluorescence quantitative polymerase chain reaction (FQ-PCR) and Western blot were used to determine the content of Bax m RNA and protein before and after transfection. The cell proliferation and apoptosis of MKN45 cells transfected with empty vector MKN45 and sh Bax were detected by BTZ gradient administration (5F2550100250500 and 1 000 nmol/L). Results A stable down-regulated MKN45-sh Bax cell line was established by transfection of Bax. After administration of BTZ gradient, the MKN45 Bax content, apoptosis degree and proliferation rate of gastric cancer cells in negative control (Mock) group were increased, and the proliferation rate was gradually decreased. The apoptosis of MKN45 cells induced by BTZ was inhibited and the proliferation ability was enhanced after silencing Bax. Conclusion Bax can partially mediate the apoptosis of gastric cancer cell line MKN45 by BTZ.
【作者单位】：中国医科大学附属盛京医院第一肿瘤科;
【基金】：国家自然科学基金(N0:81472806)
【分类号】：R735.2

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