左金丸对胃癌耐药细胞SGC-7901/DDP增殖和糖酵解的抑制作用
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摘要
目的:研究左金丸对胃癌耐药细胞SGC-7901/DDP增殖和糖酵解的影响。方法:取人胃癌细胞株SGC-7901,体外诱导顺铂(DDP)耐药细胞株SGC-7901/DDP。将耐药细胞分为空白对照组和左金丸低、中、高浓度(25、50、100μg/ml)组,各组予以相应干预。药物干预12 h、24 h、48 h后,CCK-8分别检测各组细胞存活率;药物干预24 h后,试剂盒检测各组细胞葡萄糖摄取率及细胞上清中乳酸含量,同时应用Western blot检测各组细胞中糖酵解途径关键分子C-myc、缺氧诱导因子-1α(HIF-1α)、葡萄糖转运蛋白1 (GLUT1)、乳酸脱氢酶A(LDHA)、己糖激酶Ⅱ(HKⅡ)的蛋白表达。结果:(1)与对照组比较,左金丸各浓度组在各检测时间点的细胞存活率均显著降低(P<0.01),且呈时间、浓度依赖性,提示左金丸可显著抑制SGC-7901/DDP细胞增殖。(2)药物干预24 h后,与对照组比较,左金丸各浓度组细胞的葡萄糖摄取率及乳酸含量显著降低(P<0.01),且呈浓度依赖性,提示左金丸可显著抑制SGC-7901/DDP糖酵解。(3)不同浓度左金丸干预24 h后,SGC-7901/DDP细胞中C-myc、HIF-1α、GLUT1、LDHA、HKⅡ的蛋白表达显著下调(P<0.01),且呈浓度依赖性。结论:左金丸可显著抑制胃癌耐药细胞SGC-7901/DDP的增殖,且这一过程可能与抑制糖酵解相关。
Objective: To study the effects of Zuojin Pill on the proliferation and glycolysis of cisplatin( DDP)-resistant gastric cancer cell line SGC-7901/DDP. Methods: Human gastric cancer cell line SGC-7901 was taken,and the DDP-resistant cell line SGC-7901/DDP was induced in vitro. The resistant cells were divided into the blank control group and Zuojin Pill groups with low-,medium-and high-concentration( 25,50,100 μg/ml). Each group was treated with the corresponding intervention. After treatment for 12,24 and 48 hours,CCK8 assay was used to evaluate the cell survival rate. After treatment for 24 hours,the glucose uptake and the lactic acid content in cell supernatant were detected by kit. Meanwhile,the expressions of proliferation and glycolysis related proteins,including C-myc,hypoxia-inducible factor-1α( HIF-1α),glucose transporter 1( GLUT1),lactate dehydrogenase A( LDHA),and hexokinaseⅡ( HKⅡ) were detected by Western blot. Results:(1)Compared with the control group,the survival rates of Zuojin Pill groups with different concentrations were decreased significantly at each timepoint( P<0.01),in a time-and concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the cell proliferation of SGC-7901/DDP.(2)After treatment for 24 hours,compared with the control group,the levels of glucose uptake and lactic acid content were significantly reduced in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the glycolysis of SGC-7901/DDP.(3)After treatment for 24 hours,the expressions of C-myc,HIF-1α,GLUT1,LDHA and HK Ⅱ were significantly down-regulated in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner. Conclusion: Zuojin Pill can significantly inhibit the proliferation of DDP-resistant gastric cancer cell SGC-7901/DDP,which may be related to the inhibition on glycolysis.
Objective: To study the effects of Zuojin Pill on the proliferation and glycolysis of cisplatin( DDP)-resistant gastric cancer cell line SGC-7901/DDP. Methods: Human gastric cancer cell line SGC-7901 was taken,and the DDP-resistant cell line SGC-7901/DDP was induced in vitro. The resistant cells were divided into the blank control group and Zuojin Pill groups with low-,medium-and high-concentration( 25,50,100 μg/ml). Each group was treated with the corresponding intervention. After treatment for 12,24 and 48 hours,CCK8 assay was used to evaluate the cell survival rate. After treatment for 24 hours,the glucose uptake and the lactic acid content in cell supernatant were detected by kit. Meanwhile,the expressions of proliferation and glycolysis related proteins,including C-myc,hypoxia-inducible factor-1α( HIF-1α),glucose transporter 1( GLUT1),lactate dehydrogenase A( LDHA),and hexokinaseⅡ( HKⅡ) were detected by Western blot. Results:(1)Compared with the control group,the survival rates of Zuojin Pill groups with different concentrations were decreased significantly at each timepoint( P<0.01),in a time-and concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the cell proliferation of SGC-7901/DDP.(2)After treatment for 24 hours,compared with the control group,the levels of glucose uptake and lactic acid content were significantly reduced in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner,which suggested that Zuojin Pill could significantly inhibit the glycolysis of SGC-7901/DDP.(3)After treatment for 24 hours,the expressions of C-myc,HIF-1α,GLUT1,LDHA and HK Ⅱ were significantly down-regulated in Zuojin Pill groups with different concentrations( P<0.01),in a concentration-dependent manner. Conclusion: Zuojin Pill can significantly inhibit the proliferation of DDP-resistant gastric cancer cell SGC-7901/DDP,which may be related to the inhibition on glycolysis.
引文
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[18]XIAO H, WANG J, YAN W, et al. GLUT1 regulates cell glycolysis and proliferation in prostate cancer[J]. Prostate,2018,78(2):86-94.
[19]LI H,HU S,PANG Y,et al.Bufalin inhibits glycolysis-induced cell growth and proliferation through the suppression of Integrinβ2/FAK signaling pathway in ovarian cancer[J]. Am J Cancer Res,2018,8(7):1288-1296.
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[23]ICARD P,SHUL MAN S,FARHAT D,et al. How the Warburg effect supports aggressiveness and drug resistance of cancer cells?[J]. Drug Resist Updat,2018,38:1-11.
[2]ZHITOMIRSKY B,ASSARAF Y G. Lysosomes as mediators of drug resistance in cancer[J]. Drug Resist Updat,2016,24:23-33.
[3] BHATTACHARYA B,MOHD OMAR M F, SOONG R. The Warburg effect and drug resistance[J]. Br J Pharmacol,2016,173(6):970-979.
[4]WARBURG O,WIND F,NEGELEIN E. THE METABOLISM OF TUMORS IN THE BODY[J]. J Gen Physiol,1927,8(6):519-530.
[5]CHAN A W,GILL R S,SCHILLER D,et al. Potential role of metabolomics in diagnosis and surveillance of gastric cancer[J].World J Gastroenterol,2014,20(36):12874-12882.
[6]周祥羽,邹忠杰.左金丸化学成分及现代药理研究进展[J].广东化工,2017,44(10):89-90.
[7]TANG Q F,SUN J,YU H,et al. The Zuo Jin Wan Formula Induces Mitochondrial Apoptosis of Cisplatin-Resistant Gastric Cancer Cells via Cofilin-1[J]. Evid Based Complement Alternat Med,2016,2016:8203789.
[8]TANG Q F,JI Q,QIU Y Y,et al. Synergistic Effect of ZuoJin Wan on DDP-Induced Apoptosis in Human Gastric Cancer SGC-7901/DDP Cells[J]. Evid Based Complement Alternat Med,2014,2014:724764.
[9]GUO X F,LIU J P,MA S Q,et al. Avicularin reversed multidrugresistance in human gastric cancer through enhancing Bax and BOK expressions[J]. Biomed Pharmacother,2018,103:67-74.
[10]HYUN H B,MOON J Y,CHO S K. Quercetin Suppresses CYR61-Mediated Multidrug Resistance in Human Gastric Adenocarcinoma AGS Cells[J]. Molecules,2018,23(2):209.
[11] FU Z,HAN X,DU J,et al. Euphorbia lunulata extract acts on multidrug resistant gastric cancer cells to inhibit cell proliferation,migration and invasion,arrest cell cycle progression,and induce apoptosis[J]. J Ethnopharmacol,2018,212:8-17.
[12]熊英,刘春英,王淳.参芪扶正注射液对人肺腺癌耐药细胞株A549/DDP的增敏作用[J].上海中医药大学学报,2016,30(4):52-56.
[13]张彦博,李琦,刘艳娜,等.左金丸通过Wnt/β-catenin信号通路对幽门螺杆菌感染的人胃癌细胞转移的影响[J].上海中医药大学学报,2015,29(2):44-47.
[14]AKRAM M. Mini-review on glycolysis and cancer[J]. J Cancer Educ,2013,28(3):454-457.
[15]WARBURG O. On the origin of cancer cells[J]. Science,1956,123(3191):309-314.
[16] VANDER HEIDEN M G,CANTLEY L C,THOMPSON C B.Understanding the Warburg effect:the metabolic requirements of cell proliferation[J]. Science,2009,324(5930):1029-1033.
[17]PAVLOVA N N,THOMPSON C B. The Emerging Hallmarks of Cancer Metabolism[J]. Cell Metab,2016,23(1):27-47.
[18]XIAO H, WANG J, YAN W, et al. GLUT1 regulates cell glycolysis and proliferation in prostate cancer[J]. Prostate,2018,78(2):86-94.
[19]LI H,HU S,PANG Y,et al.Bufalin inhibits glycolysis-induced cell growth and proliferation through the suppression of Integrinβ2/FAK signaling pathway in ovarian cancer[J]. Am J Cancer Res,2018,8(7):1288-1296.
[20] ZHANG X,LI Y,MA Y,et al. Yes-associated protein(YAP)binds to HIF-1αand sustains HIF-1αprotein stability to promote hepatocellular carcinoma cell glycolysis under hypoxic stress[J]. J Exp Clin Cancer Res,2018,37(1):216.
[21]WARFEL N A, EL-DEIRY W S. HIF-1 signaling in drug resistance to chemotherapy[J]. Curr Med Chem,2014,21(26):3021-3028.
[22]邓堂刚,周宇,冯佩富,等.泛素-蛋白酶体通路介导细胞内cMyc调控的研究进展[J].中国细胞生物学学报,2017,39(4):488-495.
[23]ICARD P,SHUL MAN S,FARHAT D,et al. How the Warburg effect supports aggressiveness and drug resistance of cancer cells?[J]. Drug Resist Updat,2018,38:1-11.