發(fā)布時間：2018-06-14 16:21

  本文選題：人卵巢細(xì)胞 + 缺氧�。� 參考：《中國藥房》2017年13期

【摘要】：目的:研究缺氧條件下人卵巢癌細(xì)胞(SKOV3)對順鉑耐藥與Toll樣受體9(TLR9)的關(guān)系。方法:采用免疫熒光法檢測破膜和未破膜SKOV3中TLR9的表達(dá)。取SKOV3,加入TLR9特異性激動劑Cp G-ODN 2006(A組)及其同型對照Cp G-ODN 2006control(B組)作用6、12、24 h后,加入順鉑,以CCK-8法檢測細(xì)胞抑制率。取SKOV3或經(jīng)0(未加)、1、10、102、103μmol/L TLR9特異性拮抗劑氯喹預(yù)處理3 h后的SKOV3,加入SKOV3常氧(21%O2)、缺氧(1%O2)孵育6、12、24 h的細(xì)胞上清作用24 h后,加入順鉑,檢測細(xì)胞增殖抑制率,計算預(yù)處理細(xì)胞缺氧時藥物抑制率的降低倍數(shù)(HICR);并采用Western blot法檢測常氧24 h細(xì)胞上清(C組)、缺氧24 h細(xì)胞上清(D組)、缺氧24 h細(xì)胞上清+10μmol/L氯喹(E組)條件下SKOV3中多藥耐藥相關(guān)蛋白(MRP)的表達(dá)。結(jié)果:TLR9在SKOV3的細(xì)胞膜和細(xì)胞質(zhì)中均有表達(dá)。與A組比較,B組細(xì)胞增殖抑制率降低(P0.01)。與常氧細(xì)胞上清比較,缺氧細(xì)胞上清作用后細(xì)胞增殖抑制率降低(P0.05)。與未加氯喹比較,加入10、102μmol/L氯喹能明顯降低HICR(P0.01),其中缺氧24 h細(xì)胞上清+10μmol/L氯喹降低最明顯。與C組比較,D組細(xì)胞中MRP表達(dá)增強(qiáng)(P0.01);與D組比較,E組細(xì)胞中MRP表達(dá)減弱(P0.01)。結(jié)論:TLR9受體在缺氧下可被激活,并可導(dǎo)致SKOV3對順鉑耐藥,該作用可能與上調(diào)MRP表達(dá)有關(guān)。
[Abstract]:Objective: To study the relationship between cisplatin resistance and Toll like receptor 9 (TLR9) in human ovarian cancer cells (SKOV3) under hypoxic conditions. Methods: the expression of TLR9 in the rupture and unbroken membrane SKOV3 was detected by immunofluorescence. SKOV3, TLR9 specific agonist Cp G-ODN 2006 (A group) and the same type control Cp G-ODN (Group) were added. The inhibition rate of cisplatin was detected by CCK-8. SKOV3 or 0 (not added), chloroquine, a specific antagonist of 1,10102103 mu mol/L TLR9, was pretreated with chloroquine, SKOV3, SKOV3 normal oxygen (21%O2), and hypoxia (1%O2) incubated for 24 of the cell supernatant of 6,12,24 h, and cisplatin was added to detect cell proliferation inhibition rate and to calculate the drug pretreated cells when hypoxia. The inhibition rate was reduced (HICR), and Western blot method was used to detect the expression of multidrug resistance associated protein (C), hypoxia 24 h cell supernatant (group C), hypoxia 24 h cell supernatant (group D), and +10 micron mol/L chloroquine (E group) under the condition of hypoxia (E group). The inhibitory rate of cell proliferation decreased (P0.01). Compared with the supernatant of normoxic cells, the inhibition rate of cell proliferation decreased (P0.05). Compared with chloroquine, 10102 mol/L chloroquine could obviously decrease HICR (P0.01), and the decrease of +10 u mol/L chloroquine in 24 h cell supernatant was the most obvious. Compared with group C, MRP expression in the D group was enhanced. P0.01); compared with group D, the expression of MRP in E group was weakened (P0.01). Conclusion: TLR9 receptor can be activated under hypoxia and may lead to SKOV3 resistance to cisplatin, which may be related to the up regulation of MRP expression.
【作者單位】： 第三軍醫(yī)大學(xué)第三附屬醫(yī)院藥劑科;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(No.30973493)
【分類號】：R737.31
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本文編號：2018123