中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
4期
757-759
,共3页
癌,肝细胞%索拉非尼%耐药性
癌,肝細胞%索拉非尼%耐藥性
암,간세포%색랍비니%내약성
Carcinoma,hepatocellular%Sorafenib%Drug resistance
目的 建立索拉非尼诱导肝癌耐药细胞株并研究耐药细胞的功能特性.方法 采用药物连续诱导法建立索拉非尼耐药细胞株模型;流式细胞术检测细胞凋亡率差异;实时定量聚合酶链反应(Real-time PCR)检测多药耐药(MDR1)基因表达变化;Western blot检测蛋白激酶B(Akt)和细胞外信号调节激酶(ERK1/2)磷酸化水平差异.结果 耐药组和空白组比较:其细胞生存率(％)显著升高,差异有统计学意义(SK-HEP-1:61.73比30.92;HUH-7:54.14比20.73,P＜0.05);细胞凋亡率(％)显著下降,差异有统计学意义(SK-HEP-1:42.40比21.17;HUH-7:48.23比17.63,P＜0.05);MDR1耐药基因表达显著增加,差异有统计学意义(SK-HEP-1:10.87比1.59;HUH-7:9.58比1.23,P＜0.05);并且耐药组细胞Akt和ERK1/2磷酸化水平上调.结论 体外成功建立索拉非尼诱导的HUH-7和SK-HEP-1耐药细胞株,其耐药机制与Akt和ERK1/2磷酸化水平上调导致耐药基因MDR1表达增加相关.
目的 建立索拉非尼誘導肝癌耐藥細胞株併研究耐藥細胞的功能特性.方法 採用藥物連續誘導法建立索拉非尼耐藥細胞株模型;流式細胞術檢測細胞凋亡率差異;實時定量聚閤酶鏈反應(Real-time PCR)檢測多藥耐藥(MDR1)基因錶達變化;Western blot檢測蛋白激酶B(Akt)和細胞外信號調節激酶(ERK1/2)燐痠化水平差異.結果 耐藥組和空白組比較:其細胞生存率(％)顯著升高,差異有統計學意義(SK-HEP-1:61.73比30.92;HUH-7:54.14比20.73,P＜0.05);細胞凋亡率(％)顯著下降,差異有統計學意義(SK-HEP-1:42.40比21.17;HUH-7:48.23比17.63,P＜0.05);MDR1耐藥基因錶達顯著增加,差異有統計學意義(SK-HEP-1:10.87比1.59;HUH-7:9.58比1.23,P＜0.05);併且耐藥組細胞Akt和ERK1/2燐痠化水平上調.結論 體外成功建立索拉非尼誘導的HUH-7和SK-HEP-1耐藥細胞株,其耐藥機製與Akt和ERK1/2燐痠化水平上調導緻耐藥基因MDR1錶達增加相關.
목적 건립색랍비니유도간암내약세포주병연구내약세포적공능특성.방법 채용약물련속유도법건립색랍비니내약세포주모형;류식세포술검측세포조망솔차이;실시정량취합매련반응(Real-time PCR)검측다약내약(MDR1)기인표체변화;Western blot검측단백격매B(Akt)화세포외신호조절격매(ERK1/2)린산화수평차이.결과 내약조화공백조비교:기세포생존솔(％)현저승고,차이유통계학의의(SK-HEP-1:61.73비30.92;HUH-7:54.14비20.73,P＜0.05);세포조망솔(％)현저하강,차이유통계학의의(SK-HEP-1:42.40비21.17;HUH-7:48.23비17.63,P＜0.05);MDR1내약기인표체현저증가,차이유통계학의의(SK-HEP-1:10.87비1.59;HUH-7:9.58비1.23,P＜0.05);병차내약조세포Akt화ERK1/2린산화수평상조.결론 체외성공건립색랍비니유도적HUH-7화SK-HEP-1내약세포주,기내약궤제여Akt화ERK1/2린산화수평상조도치내약기인MDR1표체증가상관.
Objective To establish sorafenib-induced drug-resistant hepatoma cell lines and study the functional properties of resistant cells.Methods By using continuity induction in vitro and the obtained 50％ inhibitory dose (IC5o) value,we established drug-resistant hepatoma cell lines.Apoptosis rate between different cell groups was examined by flow cytometry.The expression of multi-drug resistance gene (MDR1) was examined by real-time quantitative polymerase chain reaction (Real-time PCR).Western blotting analysis was applied to examine the phosphorylation level of protein kinase B (Akt) and extracellular signal-regulated kinases (ERK1/2).Results Sorafenib-resistant cell lines had a longer survival rate than control cells (％) (SK-HEP-1:61.75 vs.30.92 ; HUH-7:54.14 vs.20.73,P ＜ 0.05).As compared with control cells,apoptosis rate (％) (SK-HEP-1:42.40 vs.21.17; HUH-7:48.23 vs.17.63) and MDR1 gene expression in drug-resistant cell lines were decreased significantly (SK-HEP-1:10.87 vs.1.59; HUH-7:9.58 vs.1.23,P ＜0.05).Furthermore,sorafenib was found to induce the phosphorylation of Akt and ERK1/2.Conclusion HUH-7 and SK-HEP-1 sorafenib-resistant cell lines were successfully established.The underlying mechanism may be associated with sorafenib-induced Akt and ERK1/2 activation that leads to high expression of MDR1.
