RNA结合蛋白38通过增加人表皮生长因子受体2的表达诱导乳腺癌BT474细胞对曲妥珠单抗的敏感性

目的探讨敲除和过表达外源性RNA结合蛋白38（RNPC1）基因后，人表皮生长因子受体2（HER-2）阳性乳腺癌细胞对曲妥珠单抗敏感性的变化。方法采用慢病毒转染法敲除和过表达RNPC1基因，采用实时荧光定量PCR法检测各组BT474细胞中RNPC1和HER-2mRNA的表达，Western blot法检测RNPC1和HER-2蛋白以及P13K／AKT蛋白的表达。以不同浓度的曲妥珠单抗处理各组BT474细胞，采用流式细胞术检测各组BT474细胞的凋亡率，细胞计数试剂盒8（CCK-8）法检测生长抑制率。以20μg/ml曲妥珠单抗处理各组BT474细胞，采用Western blot法检测各组BT474...

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Published in中华肿瘤杂志 Vol. 38; no. 3; pp. 172 - 178
Main Author 李春莲 周旭婕 娄培培 夏添松 石靓 王莹 丁强
Format Journal Article
LanguageChinese
Published 210029南京医科大学第一附属医院乳腺外科 2016
Subjects
RNA结合蛋白38
乳腺肿瘤
人表皮生长因子受体2
凋亡
增殖
人表皮生长因子受体2
乳腺肿瘤
RNA结合蛋白38
增殖
Human epidermal growth factor receptor-2
Subject words] Breast neoplasms
RNA binding protein 38
Proliferation
凋亡
Apoptosis
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Summary:目的探讨敲除和过表达外源性RNA结合蛋白38（RNPC1）基因后，人表皮生长因子受体2（HER-2）阳性乳腺癌细胞对曲妥珠单抗敏感性的变化。方法采用慢病毒转染法敲除和过表达RNPC1基因，采用实时荧光定量PCR法检测各组BT474细胞中RNPC1和HER-2mRNA的表达，Western blot法检测RNPC1和HER-2蛋白以及P13K／AKT蛋白的表达。以不同浓度的曲妥珠单抗处理各组BT474细胞，采用流式细胞术检测各组BT474细胞的凋亡率，细胞计数试剂盒8（CCK-8）法检测生长抑制率。以20μg/ml曲妥珠单抗处理各组BT474细胞，采用Western blot法检测各组BT474细胞中凋亡相关蛋白的表达。结果实时荧光定量PCR和Western blot法检测结果均显示，RNPC1过表达后，RNPC1和HER2mRNA及蛋白的表达均增高；而敲除RNPC1基因后，RNPC1和HER2mRNA及蛋白的表达均降低。RNPC1的过表达能减少BT474细胞中p-P13K和p-AKT蛋白的表达；RNPC1的敲除能增加p-P13K和p-AKT蛋白的表达。5、10、15、20和25μg／ml曲妥珠单抗处理敲除实验组BT474细胞48h后的生长抑制率分别为（9．67±1．18）％、（21．67±1．23）％、（30．33±1．25）％、（40．33±1．69）％和（53．00±1．63）％，均明显低于相应浓度曲妥珠单抗处理的敲除对照组[分别为（14．00±0．82）％、（27．67±1．25）％、（39．67±1．79）％、（53．67±1．50）％和（63．33±1．52）％，均P〈0．05]。5、10、15、20和25μg／ml曲妥珠单抗处理过表达实验组BT474细胞48h后的生长抑制率分别为（20．33±1．25）％、（35．38±2．05）％、（50．43±2．12）％、（65．35±2．08）％和（76．00±2．16）％，均明显高于相应浓度曲妥珠单抗处理的过表达对照组[分别为（13．67±1．24）％、（27．86±2．05）％、（39．72±1．69）％、（53．33±1．70）％和（62．68±2．07）％，均P〈0．05]。10、20和30μg/ml曲妥珠单抗处理敲除实验组BT474细胞48h后的凋亡率分别为（11．64±0．68）％、（16．60±1．01）％和（25．14±3．12）％，均明显低于
Bibliography:Objective To explore the sensitivity of HER-2-positive BT474 breast cancer cells to trastuzumab after knockdown or overexpression of RNA binding protein 38 （RNPC1）. Methods The expressions of RNPC1 and HER-2 mRNA were detected by qRT-PCR, and the expressions of RNPC1, HER-2 and PI3K/AKT proteins were detected by Western blot after transfected with RNPC1 lentiviral vector, respectively. The experimental groups were treated with different concentration of trastuzumab, and cell apoptosis rate was analyzed by 7-AAD/APC double staining flow cytometry, and cell growth inhibition rate was tested by cell counting kit 8 （CCK-8）. The expression of apoptosis-related proteins was detected by Western blot assay. Results The results of qRT-PCR showed that overexpression of RNPC1 increased the expressions of RNPC1 and HER-2 mRNA, and the expressions of RNPC1 and HER-2 were decreased after RNPC1 knockdown. The knockdown of RNPC1 decreased the expressions of RNPC1 and HER-2. Moreover, overexpression of RNPC1 decreased and kno
ISSN:0253-3766
DOI:10.3760/cma.j.issn.0253-3766.2016.03.003