姜黄素对胰腺癌SW1990细胞耐吉西他滨的逆转作用及机制研究
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篇名: 姜黄素对胰腺癌SW1990细胞耐吉西他滨的逆转作用及机制研究
TITLE:
摘要: 目的:研究姜黄素对胰腺癌SW1990细胞耐吉西他滨(GEM)的逆转作用及机制。方法:采用CCK8法检测不同浓度(50、100、150、200、250 μmol/L)GEM对SW1990细胞和耐GEM SW1990细胞(SW1990/GEM耐药细胞)存活率的影响,计算半数抑制浓度(IC50)和耐药倍数;采用CCK8法检测不同浓度(1、5、10、20、40 μmol/L)姜黄素对SW1990/GEM耐药细胞存活率的影响,计算IC50;采用CCK8法检测2.41 μmol/L姜黄素与不同浓度(25、50、75、100、125 μmol/L)GEM联用对SW1990细胞和SW1990/GEM耐药细胞存活率的影响,计算GEM的IC50和耐药逆转倍数。采用流式细胞仪检测以GEM的IC50为给药浓度,GEM单用、姜黄素(2.41 μmol/L)与GEM联用处理SW1990细胞和SW1990/GEM耐药细胞后的细胞凋亡率和细胞周期分布,并采用Western blot法检测细胞中脂肪酸合成酶(FAS)、蛋白激酶B(AKT)、磷酸化AKT(p-AKT)、磷脂酰肌醇激酶(PI3K)、磷酸化PI3K(p-PI3K)、胱天蛋白酶3(Caspase-3)、B淋巴细胞瘤2(Bcl-2)及其相关X蛋白(Bax)的蛋白表达,反转录聚合酶链式反应(RT-PCR)检测mRNA表达。结果:GEM对SW1990细胞的IC50为92 μmol/L,对SW1990/GEM耐药细胞的IC50为216 μmol/L,SW1990细胞对GEM的耐药倍数为2.35;姜黄素对SW1990/GEM耐药细胞的IC50为9.2 μmol/L;2.41 μmol/L姜黄素下,GEM对SW1990细胞的IC50为75 μmol/L,对SW1990/GEM耐药细胞的IC50为98 μmol/L,SW1990/GEM耐药细胞对GEM的耐药逆转倍数为2.2。与GEM单用比较,姜黄素与GEM联用时SW1990细胞和SW1990/GEM耐药细胞的凋亡率均明显升高(P<0.05),主要阻滞在G0/G1期,细胞中FAS、p-AKT、p-PI3K、Bcl-2蛋白表达水平和FAS、Bcl-2 mRNA表达水平均明显降低(P<0.05),Bax、Cascapse-3蛋白表达水平和mRNA表达水平均明显升高(P<0.05)。结论:姜黄素可逆转SW1990细胞对GEM的耐药性,其机制可能与PI3K/AKT通路有关。
ABSTRACT: OBJECTIVE: To study the effects of curcumin on gemcitabine (GEM)-resistant pancreatic cancer SW1990 cells and its mechanism. METHODS: CCK8 assay was used to detect the effects of different concentrations of GEM (50, 100, 150, 200, 250 μmol/L) on the survival rate of SW1990 cells and GEM-resistant SW1990 cells (SW1991/GEM resistant cells); half inhibitory concentration (IC50) and drug resistance multiple were calculated. CCK8 assay was performed to detect the effects of different concentrations of curcumin (1, 5, 10, 20, 40 μmol/L) on survival rate of SW1990/GEM resistant cells, and IC50 was calculated. CCK8 assay was used to detect the effects of curcumin 2.41 μmol/L combined with different concentrations of GEM (25, 50, 75, 100, 125 μmol/L) on the survival rate of SW1990 cells and SW1990/GEM resistant cells, and IC50 and drug resistance reversal fold of GEM were calculated. Flow cytometry was carried out to detect the cell cycle distribution and apoptosis rate of SW1990 after treated with GEM alone or curcumin (2.41 μmol/L) combined with GEM using IC50 of GEM as drug concentration. Western blot assay was used to the protein expression of FAS, AKT, p-AKT, PI3K, p-PI3K, Caspase-3, Bcl-2 and related X protein (Bax). RT-PCR was used to detect mRNA expression. RESULTS: IC50 of GEM to SW1990 cells was 92 μmol/L. IC50 of GEM to SW1990/GEM resistant cells was 216 μmol/L, and drug resistance multiple SW1990 cells to GEM was 2.35. IC50 of curcumin to SW1990/GEM resistant cells was 9.2 μmol/L. Under 2.41 μmol/L curcumin, IC50 of GEM to SW1990 cells was 75  μmol/L, and IC50 of GEM to SW1990/GEM resistant cells was 98 μmol/L; drug resistance reversal multiple of SW1990/GEM resistant cells to GEM was 2.2. Compared with GEM alone, the apoptosis rate of SW1990 cells and SW1990/GEM resistant cells were increased significantly after curcumin combined with GEM (P<0.05), blocking at G0/G1 phase; the protein expression of FAS, p-AKT, p-PI3K and Bcl-2 and mRNA expression of FAS and Bcl-2 were decreased significantly  (P<0.05); the protein and mRNA expression of Bax      and Caspase-3 were increased significantly (P<0.05). CONCLUSIONS: Curcumin can reverse drug resistance of SW1990 cells to GEM, the mechanism of which may be associated with PI3K/AKT pathway.
期刊: 2019年第30卷第9期
作者: 彭梦媛,邱峰,黄丹,秦霞,张渊
AUTHORS: PENG Mengyuan,QIU Feng,HUANG Dan,QIN Xia,ZHANG Yuan
关键字: 姜黄素;吉西他滨;胰腺癌SW1990细胞;耐药;凋亡;机制
KEYWORDS: Curcumin; Gemcitabine; Pancreatic cancer SW1990 cell; Drug resistance; Apoptosis; Mechanism
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