GCS overexpression is associated with multidrug resistance of human HCT-8 colon cancer cells.
GCS overexpression is associated with multidrug resistance of human HCT-8 colon cancer cells.
复制标题
GCS过表达与人HCT-8结肠癌细胞的多药耐药性相关
DOI：
10.1186/1756-9966-31-23
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发表时间：
2012-03-16
期刊：
影响因子：
--
通讯作者：
Yang G
中科院分区：
文献类型：
--
作者：
Song M;Zang W;Zhang B;Cao J;Yang G
多药耐药是结肠癌治疗成功的主要障碍之一。葡萄糖神经酰胺合酶（Glucosylceramide synthase，GCS）与多药耐药（multidrug resistance，MDR）有关，可降低神经酰胺水平，帮助细胞逃避神经酰胺诱导的细胞凋亡。方法采用CCK-8细胞计数试剂盒检测细胞增殖和细胞毒性。半定量RT-PCR检测GCS、MDR 1 mRNA水平，Western blot检测GCS、caspase-3和P-gp蛋白水平。结果HCT-8、HCT-8/VCR、HCT-8/VCR-sh-mock和HCT-8/VCR-sh-GCS中GCS的相对mRNA水平分别为71.4 ± 1.1%、95.1 ± 1.2%、98.2 ± 1.5%和66.6 ± 2.1%。MDR 1 mRNA表达水平分别为61.3 ± 1.1%、90.5 ± 1.4%、97.6 ± 2.2%和56.1 ± 1.2%。顺铂复合物在HCT-8、HCT-8/VCR、HCT-8/VCR-sh-mock和HCT-8/VCR-sh-GCS中的IC 50分别为69.070 ± 0.253 μg/ml、312.050 ± 1.46 μ g/ml、328.741 ± 5.648 μg/ml和150.792 ± 0.967 μg/ml。Caspase-3蛋白表达水平分别为34.2 ± 0.6%、93.0 ± 0.7%、109.09 ± 0.7%、42.7 ± 1.3%。细胞凋亡率分别为8.77 ± 0.14%、12.75 ± 0.54%、15.39 ± 0.41%和8.49 ± 0.23%。
PurposeMultidrug resistance is one of the main impediments to the successful treatment of colon cancer. Glucosylceramide synthase (GCS) which is related to multidrug resistance (MDR) can reduce the level of ceramide and can help cells escape from the ceramide-induced cell apoptosis. However, the underlying mechanism is still unclear.MethodsThe cell proliferation and cell toxicity were measured with Cell Counting Kit-8 (CCK-8). The mRNA levels of GCS and MDR1 were detected by semiquantitative reverse transcription-PCR amplification, the protein levels of GCS, caspase-3 and P-gp proteins were indicated by Western blotting. The apoptosis rates of cells were measured with flow cytometry.ResultsThe relative mRNA levels of GCS in HCT-8, HCT-8/VCR, HCT-8/VCR- sh-mock and HCT-8/VCR-sh-GCS were 71.4 ± 1.1%, 95.1 ± 1.2%, 98.2 ± 1.5%, and 66.6 ± 2.1% respectively. The mRNA levels of MDR1 were respectively 61.3 ± 1.1%, 90.5 ± 1.4%, 97.6 ± 2.2% and 56.1 ± 1.2%. The IC50 of Cisplatin complexes were respectively 69.070 ± 0.253 μg/ml, 312.050 ± 1.46 μg/ml, 328.741 ± 5.648 μg/ml, 150.792 ± 0.967 μg/ml in HCT-8, HCT-8/VCR, HCT-8/VCR-sh-mock and HCT-8/VCR-sh-GCS. The protein levels of caspase-3 were 34.2 ± o.6%, 93.0 ± 0.7%, 109.09 ± 0.7%, 42.7 ± 1.3% respectively. The apoptosis rates of cells were 8.77 ± 0.14%, 12.75 ± 0.54%, 15.39 ± 0.41% and 8.49 ± 0.23% respectively.ConclusionIn conclusion, our research indicated that suppression of GCS restores the sensitivity of multidrug resistance colon cancer cells to drug treatment.
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影响因子：
37.3
作者：
Liu YY;Gupta V;Patwardhan GA;Bhinge K;Zhao Y;Bao J;Mehendale H;Cabot MC;Li YT;Jazwinski SM
通讯作者：
Jazwinski SM
影响因子：
5.8
作者：
Patwardhan, Gauri;Gupta, Vineet;Huang, Juowen;Gu, Xin;Liu, Yong-Yu
通讯作者：
Liu, Yong-Yu
影响因子：
7.2
作者：
Chauhan, D;Anderson, KC
通讯作者：
Anderson, KC
影响因子：
20.3
作者：
Ruefli, AA;Smyth, MJ;Johnstone, RW
通讯作者：
Johnstone, RW
影响因子：
4.8
作者：
Lavie, Y;Cao, HT;Cabot, MC
通讯作者：
Cabot, MC