虫草素对人胃癌细胞系HGC-27的影响及分子机制
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摘要
目的探讨虫草素对HGC-27细胞增殖、运动性和凋亡的影响及相关机制。方法用不同浓度虫草素处理人胃癌细胞系HGC-27、AGS和MGC-803,利用CCK-8法检测细胞活力,集落形成试验检测细胞增殖能力,流式细胞术检测细胞凋亡情况,体外划痕试验检测细胞运动性。Western blot检测Bcl-2、Bax、ERK、磷酸化ERK (p-ERK)和糖原合成酶激酶-3β(GSK-3β)蛋白表达水平。结果与对照组相比,虫草素处理组明显抑制细胞增殖、集落形成率、细胞运动性并诱导细胞凋亡,下调Bcl-2、GSK-3β和p-ERK蛋白表达水平,且呈浓度和时间依赖性,但对Bax蛋白表达水平没有影响。结论虫草素通过调节ERK/GSK-3β信号通路起到抗肿瘤作用,有望成为临床干预和治疗胃癌的潜在药物。
OBJECTIVE To investigate the effects and the related molecular mechanisms of cordycepin on cell proliferation, motility and apoptosis of human gastric carcinoma cell lines HGC-27. METHODS Human gastric carcinoma cell lines HGC-27, AGS and MGC-803 were treated with different doses of cordycepin. Cell counting kit-8 assay was used to determine relative cell vitality. Cell proliferation ability was assessed by colony formation assay. Cell apoptosis was analyzed by flow cytometry and cell motility was detected by healing wound assay. Western blot was used to test the expression levels of Bcl-2, Bax, ERK, p-ERK and GSK-3β. RESULTS Compared with the control group, cordycepin treatment groups could significantly inhibit cell proliferation, the rate of colony formation and cell motility, induce cell apoptosis and down-regulate the expression levels of Bcl-2, GSK-3β and p-ERK in a dose and time-dependent manner, but it could not affect the expression level of Bax. CONCLUSION Cordycepin exert significant anticancer effects by modulating ERK/GSK-3β signaling pathway, which may be potential drug for cancer prevention and therapy of gastric carcinoma in the clinic.
OBJECTIVE To investigate the effects and the related molecular mechanisms of cordycepin on cell proliferation, motility and apoptosis of human gastric carcinoma cell lines HGC-27. METHODS Human gastric carcinoma cell lines HGC-27, AGS and MGC-803 were treated with different doses of cordycepin. Cell counting kit-8 assay was used to determine relative cell vitality. Cell proliferation ability was assessed by colony formation assay. Cell apoptosis was analyzed by flow cytometry and cell motility was detected by healing wound assay. Western blot was used to test the expression levels of Bcl-2, Bax, ERK, p-ERK and GSK-3β. RESULTS Compared with the control group, cordycepin treatment groups could significantly inhibit cell proliferation, the rate of colony formation and cell motility, induce cell apoptosis and down-regulate the expression levels of Bcl-2, GSK-3β and p-ERK in a dose and time-dependent manner, but it could not affect the expression level of Bax. CONCLUSION Cordycepin exert significant anticancer effects by modulating ERK/GSK-3β signaling pathway, which may be potential drug for cancer prevention and therapy of gastric carcinoma in the clinic.
引文
[1]CATALANO V,LABIANCA R,BERETTA G D,et al.Gastric cancer[J].Crit Rev Oncol Hematol,2009,71(2):127-164.
[2]ASHTARI S,VAHEDI M.Economic burden of gastrointestinal cancer:estimation and importance[J].Int JGastrointest Cancer,2014,3(4):178-181.
[3]TULI H S,SHARMA A K,SANDHU S S,et al.Cordycepin:a bioactive metabolite with therapeutic potential[J].Life Sci,2013,93(23):863-869.
[4]LIAO Y H,LING J Y,ZHANG G Y,et al.Cordycepin induces cell cycle arrest and apoptosis by inducing DNAdamage and up-regulation of p53 in Leukemia cells[J].Cell Cycle,2015,14(5):761-771.
[5]TAO X D,NING Y,ZHAO X W,et al.The effects of cordycepin on the cell proliferation,migration and apoptosis in human lung cancer cell lines A549 and NCI-H460[J].J Pharm Pharmacol,2016,68(7):901-911.
[6]ZHOU Y L,GUO Z H,MENG Q F,et al.Cordycepin affects multiple apoptotic pathways to mediate hepatocellular carcinoma cell death[J].Anticancer Agents Med Chem,2017,17(1):143-149.
[7]WANG D,ZHANG Y F,LU J H,et al.Cordycepin,a natural antineoplastic agent,induces apoptosis of breast cancer cells via caspase-dependent pathways[J].Nat Prod Commun,2016,11(1):63-68.
[8]LEE H H,PARK C,JEONG J W,et al.Apoptosis induction of human prostate carcinoma cells by cordycepin through reactive oxygen species-mediated mitochondrial death pathway[J].Int J Oncol,2013,42(3):1036-1044.
[9]NASSER M I,MASOOD M,WEI W,et al.Cordycepin induces apoptosis in SGC-7901 cells through mitochondrial extrinsic phosphorylation of PI3K/Akt by generating ROS[J].Int J Oncol,2017,50(3):911-919.
[10]TODT F,CAKIR Z,REICHENBACH F,et al.Differential retrotranslocation of mitochondrial Bax and Bak[J].Embo J,2015,34(1):67-80.
[11]REN L P,LI X J,JIN S J.Influence of sophoridine on proliferation and caspase-3/Bcl-2/Bax signaling pathway of human pancreatic cancer cell line capan-1 cells[J].Chin JMod Appl Pharm(中国现代应用药学),2017,34(3):325-328.
[12]KHODAPASAND E,JAFARZADEH N,FARROKHI F,et al.Is Bax/Bcl-2 ratio considered as a prognostic marker with age and tumor location in colorectal cancer?[J].Iran Biomed J,2015,19(2):69-75.
[13]YE Y,CHOU G X,WANG H,et al.Effects of sesquiterpenes isolated from largehead Atractylodes Rhizome on growth,migration,and differentiation of B16 melanoma cells[J].Integr Cancer Ther,2011,10(1):92-100.
[14]ZHU Y,TAO M,WU J,et al.Adenovirus-directed expression of TIPE2 suppresses gastric cancer growth via induction of apoptosis and inhibition of AKT and ERK1/2 signaling[J].Cancer Gene Ther,2016,23(4):98-106.
[15]WANG Y,MO H M,GU J,et al.Cordycepin induces apoptosis of human acute monocytic leukemia cells via downregulation of the ERK/Akt signaling pathway[J].Exp Ther Med,2017,14(4):3067-3073.
[16]WANG J H,PARK S J,KO W G,et al.Cordycepin induces human lung cancer cell apoptosis by inhibiting nitric oxide mediated ERK/Slug signaling pathway[J].Am J Cancer Res,2017,7(3):417-432.
[17]WU J,ZHANG H T,XU C,et al.TIPE2 functions as a metastasis suppressor via negatively regulatingβ-catenin through activating GSK3βin gastric cancer[J].Int J Oncol,2016,48(1):199-206.
[18]YE Y,CHAO X J,WU J F,et al.ERK/GSK3βsignaling is involved in atractylenolide I-induced apoptosis and cell cycle arrest in melanoma cells[J].Oncol Rep,2015,34(3):1543-1548.
[2]ASHTARI S,VAHEDI M.Economic burden of gastrointestinal cancer:estimation and importance[J].Int JGastrointest Cancer,2014,3(4):178-181.
[3]TULI H S,SHARMA A K,SANDHU S S,et al.Cordycepin:a bioactive metabolite with therapeutic potential[J].Life Sci,2013,93(23):863-869.
[4]LIAO Y H,LING J Y,ZHANG G Y,et al.Cordycepin induces cell cycle arrest and apoptosis by inducing DNAdamage and up-regulation of p53 in Leukemia cells[J].Cell Cycle,2015,14(5):761-771.
[5]TAO X D,NING Y,ZHAO X W,et al.The effects of cordycepin on the cell proliferation,migration and apoptosis in human lung cancer cell lines A549 and NCI-H460[J].J Pharm Pharmacol,2016,68(7):901-911.
[6]ZHOU Y L,GUO Z H,MENG Q F,et al.Cordycepin affects multiple apoptotic pathways to mediate hepatocellular carcinoma cell death[J].Anticancer Agents Med Chem,2017,17(1):143-149.
[7]WANG D,ZHANG Y F,LU J H,et al.Cordycepin,a natural antineoplastic agent,induces apoptosis of breast cancer cells via caspase-dependent pathways[J].Nat Prod Commun,2016,11(1):63-68.
[8]LEE H H,PARK C,JEONG J W,et al.Apoptosis induction of human prostate carcinoma cells by cordycepin through reactive oxygen species-mediated mitochondrial death pathway[J].Int J Oncol,2013,42(3):1036-1044.
[9]NASSER M I,MASOOD M,WEI W,et al.Cordycepin induces apoptosis in SGC-7901 cells through mitochondrial extrinsic phosphorylation of PI3K/Akt by generating ROS[J].Int J Oncol,2017,50(3):911-919.
[10]TODT F,CAKIR Z,REICHENBACH F,et al.Differential retrotranslocation of mitochondrial Bax and Bak[J].Embo J,2015,34(1):67-80.
[11]REN L P,LI X J,JIN S J.Influence of sophoridine on proliferation and caspase-3/Bcl-2/Bax signaling pathway of human pancreatic cancer cell line capan-1 cells[J].Chin JMod Appl Pharm(中国现代应用药学),2017,34(3):325-328.
[12]KHODAPASAND E,JAFARZADEH N,FARROKHI F,et al.Is Bax/Bcl-2 ratio considered as a prognostic marker with age and tumor location in colorectal cancer?[J].Iran Biomed J,2015,19(2):69-75.
[13]YE Y,CHOU G X,WANG H,et al.Effects of sesquiterpenes isolated from largehead Atractylodes Rhizome on growth,migration,and differentiation of B16 melanoma cells[J].Integr Cancer Ther,2011,10(1):92-100.
[14]ZHU Y,TAO M,WU J,et al.Adenovirus-directed expression of TIPE2 suppresses gastric cancer growth via induction of apoptosis and inhibition of AKT and ERK1/2 signaling[J].Cancer Gene Ther,2016,23(4):98-106.
[15]WANG Y,MO H M,GU J,et al.Cordycepin induces apoptosis of human acute monocytic leukemia cells via downregulation of the ERK/Akt signaling pathway[J].Exp Ther Med,2017,14(4):3067-3073.
[16]WANG J H,PARK S J,KO W G,et al.Cordycepin induces human lung cancer cell apoptosis by inhibiting nitric oxide mediated ERK/Slug signaling pathway[J].Am J Cancer Res,2017,7(3):417-432.
[17]WU J,ZHANG H T,XU C,et al.TIPE2 functions as a metastasis suppressor via negatively regulatingβ-catenin through activating GSK3βin gastric cancer[J].Int J Oncol,2016,48(1):199-206.
[18]YE Y,CHAO X J,WU J F,et al.ERK/GSK3βsignaling is involved in atractylenolide I-induced apoptosis and cell cycle arrest in melanoma cells[J].Oncol Rep,2015,34(3):1543-1548.