萝卜硫素对人胃癌细胞HGC27的抑制作用及其机制研究
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摘要
目的探讨萝卜硫素对人胃癌细胞HGC27的抑制作用及其可能的作用机制。方法以HGC27人胃癌细胞株为研究对象,设置萝卜硫素低、中、高剂量组(20、40、80μg/mL)和溶剂对照组,CCK8法测定细胞存活率,DCFH-DA法测定ROS活性,HPLC法测定ATP含量,Western blot测定Bax、Bcl-2及p53蛋白的表达水平。结果 HGC27细胞经过不同浓度萝卜硫素干预处理后,低、中、高剂量组的HGC27细胞相对存活率显著降低(P<0.05),随着剂量升高存活率降低越明显,而且还随着干预处理时间的延长持续降低;细胞中ROS和APT水平经药物干预后显著降低(P<0.05);免疫印迹结果发现Bax和p53蛋白表达水平随剂量升高显著增加,而Bcl-2蛋白表达水平随药物剂量升高显著降低(P<0.05)。结论萝卜硫素能够抑制人胃癌细胞HGC27的生长,其机制可能与萝卜硫素清除细胞内ROS、抑制细胞ATP生存、下调抗凋亡蛋白Bcl-2及上调促凋亡蛋白Bax和抑癌蛋白p53的表达有关。
Objective To investigate the inhibitory effects of sulforaphane on human gastric cancer cells(HGC27) and its possible mechanism. Methods HGC27 cells were divided into four groups:sulforaphane low dose,middle dose and high dose groups(20,40 and 80 μg/mL) and the control group. CCK8 assay was used to detect the survival rate of HGC27 cells. The levels of reactive oxygen species(ROS) and adenosine triphosphate(ATP) were measured using DCFH-DA and HPLC,respectively. Western blot was performed to analyze the expression of Bax,Bcl-2 and p53. Results With different concentrations of sulforaphane treatment,the relative survival rates of HGC27 cells decreased in a dose-and time-dependent manner(P<0.05). ROS and ATP content decreased significantly(P<0.05). The expression levels of Bax and p53 were increased,but the expression of Bcl-2 was decreased significantly(P<0.05). Conclusion Sulforaphane can inhibit the growth of HGC27 cells,and the possible mechanism may be related with inhibition of ROS and ATP generation,upregulation of Bax and p53 expression and downregulation of Bcl-2 expression.
Objective To investigate the inhibitory effects of sulforaphane on human gastric cancer cells(HGC27) and its possible mechanism. Methods HGC27 cells were divided into four groups:sulforaphane low dose,middle dose and high dose groups(20,40 and 80 μg/mL) and the control group. CCK8 assay was used to detect the survival rate of HGC27 cells. The levels of reactive oxygen species(ROS) and adenosine triphosphate(ATP) were measured using DCFH-DA and HPLC,respectively. Western blot was performed to analyze the expression of Bax,Bcl-2 and p53. Results With different concentrations of sulforaphane treatment,the relative survival rates of HGC27 cells decreased in a dose-and time-dependent manner(P<0.05). ROS and ATP content decreased significantly(P<0.05). The expression levels of Bax and p53 were increased,but the expression of Bcl-2 was decreased significantly(P<0.05). Conclusion Sulforaphane can inhibit the growth of HGC27 cells,and the possible mechanism may be related with inhibition of ROS and ATP generation,upregulation of Bax and p53 expression and downregulation of Bcl-2 expression.
引文
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[3] 马林伟,徐红涛,赵聪俐,等. 萝卜硫素对A375 黑色素瘤增殖、生长、凋亡和血管生成的影响及其机制[J]. 沈阳药科大学学报,2017,34(8):680-686.
[4] 韩玉芳,赵延兵,李俊,等. 萝卜硫素诱导肝癌Huh- 6 细胞增殖和凋亡[J]. 河南科技大学学报( 医学版),2017,35(4):256-259.
[5] 张琳,刘益娟,郑建涛,等. 胃癌患者Sox2 蛋白和CTHRC-1 蛋白的表达与肿瘤发展的相关性[J]. 基因组学与应用生物学,2018,10(9):56-58.
[6] WANG Yakun,ZHANG Xiaotian. AFP-producing gastric cancer and hepatoid gastric cancer[J]. Zhonghua Zhong Liu Za Zhi,2017,39(11):801-807.
[7] CHOI A H,KIM J,CHAO J. Perioperative chemotherapy for resectable gastric cancer:MAGIC and beyond[J]. World J Gastroenterol,2015,21(24):7343-7348.
[8] CHEN C Y,YU Z Y,CHUANG Y S,et al. Sulforaphane attenuates EGFR signaling in NSCLC cells[J]. J Biomed Sci,2015,22(12):38-46.
[9] YANAKA A. Sulforaphane enhances protection and repair of gastric mucosa against oxidative stress in vitro,and demonstrates anti-inflammatory effects on Helicobacter pylori-infected gastric mucosae in mice and human subjects[J]. Curr Pharm Des,2011,17(16):1532-1540.
[10] CHANG H W,LI R N,WANG H R,et al. Withaferin a induces oxidative stress-mediated apoptosis and DNA damage in oral cancer cells[J]. Front Physiol,2017,8(12):634-639.
[11] 孙国贵. 活性氧与肿瘤[J].癌变·畸变·突变,2011,23(11):78-80.
[12] RODIC S,VINCENT M D. Reactive Oxygen Species (ROS) are a key determinant of cancer's metabolic phenotype[J]. Int J Cancer,2017,7(1):125-133.
[13] GILL K S,FERNANDES P,O'DONVAN T R,et al. Glycolysis inhibition as a cancer treatment and its role in an anti-tumour immune response[J]. Biochim Biophys Acta,2016,1866(1):87-105.
[14] XU R H,PELICANO H,ZHOU Y. Inhibition of glycolysis in cancer cells:A novel strategy to overcome drug resistance associated with mitochondrial respiratory defect and hypoxia[J]. Cancer Res,2017,65(5):613-621.
[15] 柯于培,秦一雨,舒涛. 青蒿素联合吉西他滨对结肠癌细胞株增殖和凋亡的影响[J]. 中成药,2018,20(9):837-841.
[16] 王玲,周巧直,王晓燕,等. 阿托伐他汀调控MMP-9、Cleaved Caspase-3、Bcl-2和Bax蛋白表达对胃癌细胞增殖、周期和凋亡的影响[J]. 胃肠病学和肝病学杂志,2018,27(11):1206-1210.
[17] MIYAMOTO T,TANIKAWA C,YODSURANG V,et al. Identi-fication of a p53-repressed gene module in breast cancer cells[J]. Oncotarget,2017,8(34):55821-55836.