【摘要】：目的驗(yàn)證BCl-2相關(guān)X(Bax)蛋白在硼替佐米(BTZ)誘導(dǎo)胃癌細(xì)胞株MKN45的凋亡作用。方法以人Bax(NC_000019.10)為靶標(biāo)設(shè)計(jì)sh RNA序列,將Bax sh RNA轉(zhuǎn)染至MKN45細(xì)胞中,G418篩選耐藥克隆。熒光定量聚合酶鏈反應(yīng)和Western blot測(cè)定轉(zhuǎn)染前后Bax m RNA和蛋白含量。給予空載體對(duì)照MKN45及sh Bax轉(zhuǎn)染的MKN45細(xì)胞BTZ梯度加藥(5、25、50、100、250、500和1 000 nmol/L),檢測(cè)其細(xì)胞增殖、凋亡程度。結(jié)果通過(guò)轉(zhuǎn)染,建立Bax表達(dá)穩(wěn)定下調(diào)的MKN45-sh Bax細(xì)胞系。給予BTZ梯度加藥后,陰性對(duì)照(Mock)組胃癌細(xì)胞MKN45 Bax含量增加,凋亡程度逐漸增加,增殖率逐漸降低。沉默Bax后BTZ誘導(dǎo)的MKN45細(xì)胞凋亡受到抑制,增殖能力增強(qiáng)。結(jié)論 Bax能部分介導(dǎo)BTZ對(duì)胃癌細(xì)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.
【作者單位】： 中國(guó)醫(yī)科大學(xué)附屬盛京醫(yī)院第一腫瘤科;
【基金】：國(guó)家自然科學(xué)基金(N0:81472806)
【分類號(hào)】：R735.2

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