Twist在低氧微环境中对宫颈癌Hela细胞顺铂耐药的作用及可能机制
发布时间：2018-12-21 08:21
【摘要】：目的耐药是影响宫颈癌患者的化疗效果及预后的主要原因。探讨Twist在低氧微环境中宫颈癌Hela细胞顺铂耐药的作用及可能机制。方法将Hela细胞分无药对照组、常氧A组(不含Co Cl2)、低氧A组(加入含终浓度为150μmol/L Co Cl2的培养基),6 h后分别向常氧A组、低氧A组加入不同浓度梯度顺铂(10-3、10-4、10-5、10-6、10-7mol/L),24 h后加MTT,测IC50及生长抑制率。以二氯化钴(Co Cl2)处理Hela细胞模拟低氧微环境;MTT法筛选低氧条件下顺铂的最适作用浓度及时间;设置常氧B组、顺铂组、低氧B组、低氧顺铂组,免疫荧光法和Western blot法检测Twist、MDR1的表达情况。结果 MTT结果示:常氧A组顺铂IC50为10-5.3mol/L,低氧A组顺铂IC50为10-4.5mol/L。当顺铂浓度≥10-5mol/L时,常氧A组、低氧A组对Hela细胞抑制率较0 mol/L顺铂差异具有统计学意义(P0.05),且常氧A组与低氧A组抑制率差异有统计学意义(P0.05)。当作用时间≥24 h时,常氧A组、低氧A组对Hela细胞抑制率较0 h时差异具有统计学意义(P0.05),且常氧A组与低氧A组抑制率差异有统计学意义(P0.05)。免疫荧光结果示:顺铂组、低氧B组、低氧顺铂组Hela细胞中Twist蛋白表达(20.81±2.07、24.25±4.51、33.14±4.24)较常氧B组(13.08±2.39)显著增高(P0.05),MDR1蛋白表达(35.26±8.41、60.13±22.32、76.00±9.96)亦较常氧B组(29.57±12.80)增高(P0.05);低氧B组及低氧顺铂组Twist与MDR1表达呈正相关(r=0.639,P0.05)。Western blot结果示:顺铂组、低氧B组、低氧顺铂组Hela细胞中Twist蛋白表达(0.777±0.066、0.786±0.010、0.990±0.052)较常氧B组(0.631±0.015)增高(P0.05),MDR1蛋白表达(0.923±0.038、0.896±0.015、1.049±0.037)亦较常氧B组(0.661±0.085)增高(P0.05;低氧B组及低氧顺铂组Twist与MDR1表达呈正相关(r=0.686,P0.05;r=0.546,P0.05)。结论低氧条件下Hela细胞对顺铂的敏感性下降,可能是低氧激活Twist,上调MDR1表达从而引起宫颈癌Hela细胞耐药。
[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，
本文编号：2388654
[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，
本文编号：2388654
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