李军萍;左剑玲;王丽娜;孟晓文;

• 1:苏州大学附属第一医院

摘要(Abstract)：

目的探讨磷脂酰肌醇激酶3(PIK3C3)小干扰RNA(siRNA)-pool对大鼠PC12细胞PIK3C3及P62/SQSTM1表达的影响。方法将处于对数生长期的PC12细胞随机分为正常组、载体组、阴性对照组及siRNA 25、50、100 nmol/L组。正常组常规培养,其余组均给予载体Lipofectamine2000转染;在转染的同时,阴性对照组加入200 pmol/L FAM-siRNA;siRNA 25、50、100 nmol/L组采用siRNA-pool法分别加入含50、100、200 nmol/L siRNA。各组转染24 h时,采用MTT法检测细胞存活率,采用RT-PCR法检测PIK3C3 mRNA相对表达量;转染48 h时,采用免疫细胞化学法检测P62/SQSTM1表达。结果转染24 h时,正常组、载体组、阴性对照组及siRNA 25、50、100nmol/L组细胞存活率分别为100%、(98.4±1.7)%、(99.7±2.4)%、(104.3±2.8)%、(101.6±5.1)%、(96.9±1.9)%,组间比较P均>0.05;PIK3C3 mRNA相对表达量分别为1、1.01±0.11、0.93±0.07、0.85±0.06、0.78±0.03、0.65±0.04,siRNA 50 nmol/L组PIK3C3 mRNA相对表达量均低于正常组及载体组,siRNA 100 nmol/L组均低于其他各组,组间比较P<0.05或<0.01。正常组及siRNA 100 nmol/L组P62/SQSTM1表达光密度值分别为0.038±0.001、0.054±0.004,组间比较P<0.01。结论 PIK3C3 siRNA-pool可抑制PC12细胞PIK3C3表达,促进P62/SQSTM1蓄积。

关键词(KeyWords)： PC12细胞;小干扰RNA;磷脂酰肌醇激酶3;P62/SQSTM1

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(81471136)

作者(Author): 李军萍;左剑玲;王丽娜;孟晓文;

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DOI:

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