【摘要】：目的耐藥是影響宮頸癌患者的化療效果及預(yù)后的主要原因。探討Twist在低氧微環(huán)境中宮頸癌Hela細(xì)胞順鉑耐藥的作用及可能機(jī)制。方法將Hela細(xì)胞分無(wú)藥對(duì)照組、常氧A組(不含Co Cl2)、低氧A組(加入含終濃度為150μmol/L Co Cl2的培養(yǎng)基),6 h后分別向常氧A組、低氧A組加入不同濃度梯度順鉑(10-3、10-4、10-5、10-6、10-7mol/L),24 h后加MTT,測(cè)IC50及生長(zhǎng)抑制率。以二氯化鈷(Co Cl2)處理Hela細(xì)胞模擬低氧微環(huán)境;MTT法篩選低氧條件下順鉑的最適作用濃度及時(shí)間;設(shè)置常氧B組、順鉑組、低氧B組、低氧順鉑組,免疫熒光法和Western blot法檢測(cè)Twist、MDR1的表達(dá)情況。結(jié)果 MTT結(jié)果示:常氧A組順鉑IC50為10-5.3mol/L,低氧A組順鉑IC50為10-4.5mol/L。當(dāng)順鉑濃度≥10-5mol/L時(shí),常氧A組、低氧A組對(duì)Hela細(xì)胞抑制率較0 mol/L順鉑差異具有統(tǒng)計(jì)學(xué)意義(P0.05),且常氧A組與低氧A組抑制率差異有統(tǒng)計(jì)學(xué)意義(P0.05)。當(dāng)作用時(shí)間≥24 h時(shí),常氧A組、低氧A組對(duì)Hela細(xì)胞抑制率較0 h時(shí)差異具有統(tǒng)計(jì)學(xué)意義(P0.05),且常氧A組與低氧A組抑制率差異有統(tǒng)計(jì)學(xué)意義(P0.05)。免疫熒光結(jié)果示:順鉑組、低氧B組、低氧順鉑組Hela細(xì)胞中Twist蛋白表達(dá)(20.81±2.07、24.25±4.51、33.14±4.24)較常氧B組(13.08±2.39)顯著增高(P0.05),MDR1蛋白表達(dá)(35.26±8.41、60.13±22.32、76.00±9.96)亦較常氧B組(29.57±12.80)增高(P0.05);低氧B組及低氧順鉑組Twist與MDR1表達(dá)呈正相關(guān)(r=0.639,P0.05)。Western blot結(jié)果示:順鉑組、低氧B組、低氧順鉑組Hela細(xì)胞中Twist蛋白表達(dá)(0.777±0.066、0.786±0.010、0.990±0.052)較常氧B組(0.631±0.015)增高(P0.05),MDR1蛋白表達(dá)(0.923±0.038、0.896±0.015、1.049±0.037)亦較常氧B組(0.661±0.085)增高(P0.05;低氧B組及低氧順鉑組Twist與MDR1表達(dá)呈正相關(guān)(r=0.686,P0.05;r=0.546,P0.05)。結(jié)論低氧條件下Hela細(xì)胞對(duì)順鉑的敏感性下降,可能是低氧激活Twist,上調(diào)MDR1表達(dá)從而引起宮頸癌Hela細(xì)胞耐藥。
[Abstract]:Objective Drug resistance is the main reason that affects the effect of chemotherapy and prognosis of patients with cervical cancer. To investigate the effect of Twist on cisplatin resistance of cervical cancer Hela cells in hypoxic microenvironment and its possible mechanism. Methods Hela cells were divided into control group, normoxic group (without Co Cl2) and hypoxic group (adding medium containing 150 渭 mol/L Co Cl2 final concentration). After 6 hours, Hela cells were divided into normoxic group and normoxic group. In group A, different concentrations of Cisplatin (10-3 ~ (-4) 10 ~ (-4) 10-6 ~ (-6) ~ (-7) mol / L were added, IC50 and growth inhibition rate were measured 24 h later. Hela cells were treated with cobalt dichloride (Co Cl2) to simulate hypoxic microenvironment, the optimal concentration and time of cisplatin under hypoxia were screened by MTT method. The expression of Twist,MDR1 in normoxic group, cisplatin group, hypoxia-B group and hypoxia-cisplatin group was detected by immunofluorescence and Western blot assay. Results MTT results showed that the IC50 of cisplatin was 10-5.3 mol / L in normoxic group and 10-4.5 mol / L in hypoxia group. When the concentration of cisplatin 鈮，

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