本文選題：骨肉瘤 + 內(nèi)質(zhì)網(wǎng)應(yīng)激�。� 參考：《腫瘤》2017年09期

【摘要】：目的:觀察雷公藤紅素對(duì)人骨肉瘤HOS細(xì)胞凋亡的影響,并探討其可能的作用機(jī)制。方法 :用不同濃度(0、1.5、2.5、3.5和4.5μmol/L)的雷公藤紅素處理骨肉瘤HOS細(xì)胞24 h后,采用CCK-8法檢測(cè)細(xì)胞活性,確定最佳實(shí)驗(yàn)濃度。采用倒置相差顯微鏡觀察細(xì)胞形態(tài)變化,Hoechst33258染色后相差顯微鏡觀察細(xì)胞凋亡的形態(tài)學(xué)改變,FCM法檢測(cè)細(xì)胞凋亡率和細(xì)胞內(nèi)Ca2+濃度變化,蛋白質(zhì)印跡法檢測(cè)結(jié)合免疫球蛋白(binding immunoglobulin protein,BIP)、鈣聯(lián)蛋白(calnexin,CNX)、內(nèi)質(zhì)網(wǎng)氧化還原酶1-Lα(endoplasmic reticulum oxidoreductin 1-L alpha,Ero1-Lα)、蛋白質(zhì)二硫鍵異構(gòu)酶(protein disulfide isomerase,PDI)、肌醇依賴(lài)酶1(αinositol-requiring enzyme 1 alpha,IRE1α)、蛋白激酶樣內(nèi)質(zhì)網(wǎng)激酶(protein kinase-like endoplasmic reticulum kinase,PERK)和磷酸化PERK(phosphorylatedPERK,p-PERK)、CCAAT/增強(qiáng)子結(jié)合蛋白同源蛋白(CCAAT/enhancerbinding protein homologous protein,CHOP)、剪切型caspase-12(cleavedcaspase-12,c-caspase-12)和c-caspase-3的表達(dá)變化。結(jié)果 :不同濃度的雷公藤紅素均可抑制HOS細(xì)胞活性(P值均0.05),且呈濃度依賴(lài)性。3μmol/L雷公藤紅素處理24 h后,HOS細(xì)胞形態(tài)明顯萎縮、紊亂,漂浮細(xì)胞增多,且出現(xiàn)明顯核固縮和核碎裂等典型的細(xì)胞凋亡形態(tài)學(xué)改變。與未處理對(duì)照組相比,3μmol/L雷公藤紅素處理24 h后細(xì)胞凋亡率和細(xì)胞內(nèi)Ca2+濃度明顯升高(P值均0.05)。3μmol/L雷公藤紅素處理24 h后,內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)標(biāo)志蛋白BIP、CNX、Ero1-Lα、PDI、IRE1α和p-PERK以及內(nèi)質(zhì)網(wǎng)應(yīng)激介導(dǎo)凋亡相關(guān)蛋白CHOP、c-caspase-12和c-caspase-3的表達(dá)水平明顯升高(P值均0.05)。結(jié)論 :雷公藤紅素可誘導(dǎo)人骨肉瘤HOS細(xì)胞凋亡,其作用機(jī)制可能與內(nèi)質(zhì)網(wǎng)應(yīng)激介導(dǎo)細(xì)胞凋亡通路有關(guān)。
[Abstract]:Aim: to investigate the effect of tripterine on apoptosis of human osteosarcoma HOS cells and its possible mechanism. Methods: osteosarcoma HOS cells were treated with tripterine at different concentrations of 1.5 渭 mol / L and 4.5 渭 mol / L for 24 h. The cell activity was detected by CCK-8 assay and the optimal concentration was determined. Morphological changes of apoptosis were observed by inverted phase contrast microscope (PPM) and Hoechst33258 staining. The apoptosis rate and intracellular Ca2 concentration were detected by FCM. Western blot detection of binding immunoglobulin, calnexin immunoglobulin, endoplasmic redoxoreductase 1-L 偽 -endoplasmic reticulum oxidoreductin 1-L alpha-Ero1-L 偽, protein disulfide isomerase, inositol dependent enzyme 1 (偽 inositol-requiring enzyme 1 alpha-IRE1 偽, protein kinase-like endoplasmic reticulum kinase kinase-like endoplasmic) The expression of CCAATA / enhancer binding protein, CCAATR / Enhancer-binding protein homologous protein, Caspase-12 caspase-12cleavedcaspase-12c-caspase-12) and c-caspase-3 were detected by reticulum kinase- PERK) and phosphorylated PERKN (phosphorylated PERKA) and phosphorylated PERKN (phosphorylated PERKN). Results: Tripterygium wilfordii inhibited the activity of HOS cells in a dose-dependent manner (P < 0.05). After treated with tripterine for 24 h, the morphology of Hos cells shrank significantly, and the number of floating cells increased. Typical morphological changes of apoptosis such as nuclear pyknosis and nuclear fragmentation were also observed. Compared with the untreated control group, the apoptosis rate and intracellular Ca2 concentration of tripterine treated with 3 渭 mol/L tripterine for 24 h increased significantly (P = 0.050.30 渭 mol/L) after treatment with tripterine for 24 h. The expression of endoplasmic reticulum stress-related marker protein BIP-CNX1-L 偽, PDII-IRE1 偽 and endoplasmic reticulum stress-mediated apoptosis-associated protein CHOP-c-caspase-12 and c-caspase-3 increased significantly (P < 0.05). Conclusion: Tripterygium wilfordii can induce apoptosis of human osteosarcoma HOS cells and its mechanism may be related to endoplasmic reticulum stress-mediated apoptosis pathway.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第一醫(yī)院骨科;
【基金】：國(guó)家自然科學(xué)基金資助項(xiàng)目(編號(hào):81572634) 重慶市教育委員會(huì)研究生科研創(chuàng)新項(xiàng)目(編號(hào):CYS15141)~~
【分類(lèi)號(hào)】：R738.1

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