摘要
目的探索核受体Nur77在结肠癌细胞奥利沙铂耐药中的作用及其相关机制。方法构建空载、Nur77及其抑制物载体转染结肠癌细胞SW480,将正常结肠癌细胞SW480分为空白组与对照组、空载转染结肠癌细胞SW480设为空载组、Nur77载体转染结肠癌细胞SW480设为Nur77过表达组、Nur77抑制物载体转染结肠癌细胞SW480设为Nur77抑制组,取对数生长期各组结肠癌细胞SW480采用MTT法分别检测浓度0.5μg/mL、1.0μg/mL、2.0μg/mL、4.0μg/mL、8.0μg/mL的奥利沙铂作用下细胞增殖情况,Western blot检测各组细胞NF-κB-p65、NF-κB激酶复合体(IKK)、磷酸化IKK蛋白(p-IKK)表达水平,qRT-PCR法检测各组细胞NF-κB-p65、IKK、p-IKK mRNA表达水平。结果随着奥利沙铂浓度的增加,结肠癌细胞SW480存活率逐渐降低,且尤以Nur77抑制组存活率最低(P<0.05),Nur77过表达组生存率最高(P<0.05);转染Nur77后的结肠癌细胞内NF-κB-p65、IKK、p-IKK蛋白及其mRNA表达水平较其他各组细胞升高(P<0.05),而转染Nur77抑制物的结肠癌细胞内NF-κB-p65、IKK、p-IKK蛋白及其mRNA表达水平较其他各组细胞降低(P<0.05)。结论随着奥利沙铂浓度的增加,结肠癌细胞存活率逐渐降低,但转染Nur77的结肠癌细胞存活率却异常升高,Nur77的高表达可降低结肠癌细胞的药物敏感性;且转染Nur77的结肠癌细胞内NF-κB-p65、IKK、p-IKK表达水平也随之升高,通过增加IKK、p-IKK的表达水平,进而提高NF-κB信号通路活性可能是其降低结肠癌细胞药物敏感性的主要作用机制。
Objective To explore the role of nuclear receptor Nur77 in oxaliplatin resistance of colon cancer cells and its related mechanism.Methods Colon cancer cell SW480 was transfected with Nur77 and its inhibitor vector.Normal colon cancer cell SW480 was divided into a blank group and a control group;no-load transfected colon cancer cell SW480 was set as a no-load group;Nur77 vector transfected colon cancer cell SW480 was set as Nur77 overexpression group;and Nur77 inhibitor vector transfected colon cancer cell SW480 was set as Nur77 inhibition group.Colon cancer cells SW480 of each group in logarithmic growth stage were detected by MTT assay under the treatment of 0.5μg/mL,1.0μg/mL,2.0μg/mL,4.0μg/mL and 8.0μg/mL olisatin,respectively.The expression levels of NF-κB-p65,IKK,and p-IKK in cells of each group were detected by Western blot.The mRNA expression levels of NF-κB-p65,IKK,and p-IKK in each group were detected by qRT-PCR.Results With the increase of oxalixatin concentration,the survival rate of SW480 decreased gradually,the survival rate of the Nur77 inhibition group was the lowest(P<0.05),and the survival rate of the Nur77 overexpression group was the highest(P<0.05).The expression levels of NF-κB-p65,IKK,and P-IKK proteins and their mRNA in colon cancer cells transfected with Nur77 were significantly higher than those in other groups(P<0.05).The expression levels of NF-κB-p65,IKK,P-IKK protein and mRNA in colon cancer cells transfected with Nur77 inhibitor were significantly decreased compared with those in other groups(P<0.05).Conclusions With the increase of oxalixatin concentration,the survival rate of colon cancer cells decreased gradually,but the survival rate of colon cancer cells transfected with Nur77 increased abnormally.The high expression of Nur77 could reduce the drug sensitivity of colon cancer cells.Furthermore,the expression levels of NF-κb-p65,IKK,and p-IKK in Nur77 transfected colon cancer cells were also increased.By increasing the expression levels of IKK and p-IKK,improving the acti
作者
王迎梅
王莹
李超
WANG Yingmei;WANG Ying;LI Chao(Department of General Surgery,Daqing Longnan Hospital,Daqing 163000,China)
出处
《中国肿瘤外科杂志》
CAS
2023年第6期562-566,共5页
Chinese Journal of Surgical Oncology
