塞来昔布下调sirtuin 1表达抑制转化生长因子-β1诱导人胃癌细胞SGC-7901上皮细胞-间充质转化过程的研究
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摘要
目的:探究塞来昔布(CXB)对转化生长因子-β1(TGF-β1)诱导人胃癌细胞SGC-7901上皮细胞-间充质转化(EMT)过程的影响及其分子机制。方法:用10 ng·ml~(-1)TGF-β1诱导SGC-7901细胞建立EMT模型,将建模成功的胃癌细胞分为5组:模型组、白藜芦醇组(Res,50μmol·L~(-1))、CXB低(40μmol·L~(-1))、高(100μmol·L~(-1))剂量组、Res+CXB组(50μmol·L~(-1)+100μmol·L~(-1)),以未建模的SGC-7901细胞作为对照组,采用实时荧光定量PCR(qRT-PCR)检测SGC-7901中sirtuin 1 mRNA表达情况,分别采用MTT实验、划痕实验和transwell小室检测SGC-7901细胞增殖、迁移和侵袭情况,蛋白免疫印迹(Western Blot)检测SGC-7901细胞中钙粘附蛋白-E(E-cadherin)、波形蛋白(vimentin)表达情况。结果:模型组SGC-7901细胞呈现梭形和纺锤形,且细胞间隙变大、排列松散,提示造模成功。模型组和Res组SGC-7901细胞中sirtuin 1 mRNA表达水平、SGC-7901细胞增殖率、迁移能力、侵袭能力及vimentin表达水平显著高于对照组,E-cadherin表达水平显著低于对照组(P<0.05);与模型组相比,CXB低、高剂量组SGC-7901细胞中sirtuin 1 mRNA表达水平、SGC-7901细胞增殖率、迁移能力、侵袭能力及vimentin表达水平显著降低,E-cadherin表达水平显著升高(P<0.05);Res+CXB组SGC-7901细胞中sirtuin 1 mRNA表达水平、SGC-7901细胞增殖率、迁移能力、侵袭能力vimentin表达水平显著低于Res组,E-cadherin表达水平显著高于Res组(P<0.05)。结论:CXB能够下调SGC-7901细胞中sirtuin 1表达,进而抑制TGF-β1诱导的人胃癌细胞SGC-7901 EMT过程。
Objective:To investigate the effect of celecoxib(CXB)on SGC-7901 epithelial-mesenchymal transition(EMT)process induced by TGF-β1 in human gastric cancer cells and its molecular mechanism.Methods:SGC-7901 cells were induced by 10ng·ml~(-1)TGF-β1 to establish EMT model,and the gastric cancer cells were divided into five groups,named the model group,resveratol group(Res,50μmol·L~(-1)),CXB low dose(40μmol·L~(-1))group,CXB high dose(100μmol·L~(-1))group,Res+CXB group(50μmol·L~(-1)+100μmol·L~(-1)),and the SGC-7901 cells without modeling were used as the control group.Real-time quantitative PCR(qRT-PCR)was used to detect SGC-7901 sirtuin 1 mRNA,and the cell proliferation,migration and invasion was detected by MTT assay,scratch assay and Transwell assay,respectively,the expressions of vimentin and E-cadherin were detected by Western blot.Results:The SGC-7901 cells in the model group showed shuttle and spindle shapes,the intercellular space became larger,and the cells became loosely arranged,which indicated that the modeling was successful.The expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells in the model group and Res group were significantly higher than those in the control group,and the expression level of E-cadherin in SGC-7901 cells was significantly lower than that in the control group(P<0.05).Compared with those in the model group,the expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells was significantly decreased,and the expression level of E-cadherin in SGC-7901 cells was significantly increased(P<0.05).The expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells in Res+CXB group were significantly lower than those in Res group,and the expression level of E-cadherin was significantly higher than that in Res group(P<0.05).Conclusion:CXB can down-regulate SGC-7901 cells sirtuin 1 expression,and inhibit SGC-7901 EMT process induced by TGF-β1 in human gastric cancer cell.
Objective:To investigate the effect of celecoxib(CXB)on SGC-7901 epithelial-mesenchymal transition(EMT)process induced by TGF-β1 in human gastric cancer cells and its molecular mechanism.Methods:SGC-7901 cells were induced by 10ng·ml~(-1)TGF-β1 to establish EMT model,and the gastric cancer cells were divided into five groups,named the model group,resveratol group(Res,50μmol·L~(-1)),CXB low dose(40μmol·L~(-1))group,CXB high dose(100μmol·L~(-1))group,Res+CXB group(50μmol·L~(-1)+100μmol·L~(-1)),and the SGC-7901 cells without modeling were used as the control group.Real-time quantitative PCR(qRT-PCR)was used to detect SGC-7901 sirtuin 1 mRNA,and the cell proliferation,migration and invasion was detected by MTT assay,scratch assay and Transwell assay,respectively,the expressions of vimentin and E-cadherin were detected by Western blot.Results:The SGC-7901 cells in the model group showed shuttle and spindle shapes,the intercellular space became larger,and the cells became loosely arranged,which indicated that the modeling was successful.The expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells in the model group and Res group were significantly higher than those in the control group,and the expression level of E-cadherin in SGC-7901 cells was significantly lower than that in the control group(P<0.05).Compared with those in the model group,the expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells was significantly decreased,and the expression level of E-cadherin in SGC-7901 cells was significantly increased(P<0.05).The expression level of sirtuin 1 mRNA,the proliferation,migration,invasion and vimentin expression of SGC-7901 cells in Res+CXB group were significantly lower than those in Res group,and the expression level of E-cadherin was significantly higher than that in Res group(P<0.05).Conclusion:CXB can down-regulate SGC-7901 cells sirtuin 1 expression,and inhibit SGC-7901 EMT process induced by TGF-β1 in human gastric cancer cell.
引文
1曲云慧,余福兵.早期胃癌诊断与治疗进展[J].中华临床医师杂志:电子版,2017,11(2):292-296
2诸芳芳.血清CEA、CA19-9检测对老年胃癌诊断的临床价值[J].中国老年学,2014,34(10):2864-2865
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4惠起源,魏晓萍.上皮间质转化在肿瘤发生发展中的作用[J].中国肿瘤,2013,22(3):219-222
5 Hwang KE,Cho KH,Jeong ET,et al.Down-regulated SIRT1 by Nonsteroidal Antiinflammatory Drugs Is to Inhibit TGF-1-Induced Epithelial-Mesenchymal Transition and to Suppress Migration and Invasion in Lung Cancer[J].Chest,2015,148(4):548A-548B
6邓琳,冯定庆,凌斌.COX2促进卵巢癌细胞迁移及相关机制的研究[J].现代妇产科进展,2016,25(8):561-565
7梁巍,孙文广,姜佳佳,等.TGF-β1诱导人胃腺癌SGC-7901细胞建立EMT模型[J].医学研究杂志,2014,43(6):37-41
8 Zhao R,Gong L,Li L,et al.nm23-H1 is a negative regulator of TGF-β1-dependent induction of epithelial-mesenchymal transition[J].Exp Cell Res,2013,319(5):740-749
9 Kiesslich T,Pichler M,Neureiter D.Epigenetic controlof epithelialmesenchymal-transition in human cancer[J].Mol Clin Oncol,2013,1(1):3-11
10杨燕,邹晓平.上皮间质转化与胃癌的研究进展[J].国际消化病杂志,2014,34(5):334-337
11 Vera M,Barcia E,Negro S,et al.New celecoxib multiparticulate systems to improve glioblastoma treatment[J].Int J Pharm,2014,473(1-2):518-527
12 Zhang Q,Meng X,Zheng G,et al.Antitumor activity of celecoxib,a selective cyclooxygenase-2 inhibitor,in medullary thyroid carcinoma[J].Mol Med Rep,2014,9(2):768-772
13吕丽,李文通,杨灵,等.COX-2表达与乳腺癌EMT发生关系的研究[J].中国妇幼保健,2013,28(28):4728-4730
14李裕明,韩克强,郑璐,等.上皮间质转化标志物E-cadherin和Vimentin在原发性肝癌中的表达及临床意义[J].中华肝脏病杂志,2013,21(4):279-284
15申震,周颖,赵卫东,等.Vimentin及E-Cadherin在子宫颈癌中的表达及其与上皮细胞间质化的相关性[J].安徽医科大学学报,2013,48(5):505-508
16 El-Sayed,Mona,Elazeem A,et al.Expression of epithelial-mesenchymal transition-related markers E-cadherin and vimentin in ovarian serous carcinomas:correlation with transforming growth factor-β1expression and clinicopathological factors[J].Egyptian Journal of Pathology,2016,36(1):1-8
17 Eades G,Yao Y,Yang M,et al.miR-200a regulates SIRT1 expression and epithelial to mesenchymal transition(EMT)-like transformation in mammary epithelial cells.[J].J Biol Chem,2011,286(29):25992-26002
18郝翀.SIRT1在肝细胞癌侵袭转移中的作用机制研究[D].上海:第二军医大学硕士学位论文,2014
19 Byles V,Zhu L,Lovaas JD,et al.SIRT1 induces EMT by cooperating with EMT transcription factors and enhances prostate cancer cell migration and metastasis[J].Oncogene,2012,31(43):4619-4629
20曾薇,朱金峰,于瑞冉,等.miR-34a靶向调控Sirtuin 1表达对食管鳞癌细胞侵袭、转移的影响[J].山东医药,2017,57(24):43-45
21王金凤.白藜芦醇对PC12细胞增殖内稳态影响的研究[D].广州:华南师范大学硕士学位论文,2013
2诸芳芳.血清CEA、CA19-9检测对老年胃癌诊断的临床价值[J].中国老年学,2014,34(10):2864-2865
3向传文.浅论年龄因素对接受胃癌根治术的患者术后5年生存率的影响[J].当代医药论丛,2016,14(11):11-12
4惠起源,魏晓萍.上皮间质转化在肿瘤发生发展中的作用[J].中国肿瘤,2013,22(3):219-222
5 Hwang KE,Cho KH,Jeong ET,et al.Down-regulated SIRT1 by Nonsteroidal Antiinflammatory Drugs Is to Inhibit TGF-1-Induced Epithelial-Mesenchymal Transition and to Suppress Migration and Invasion in Lung Cancer[J].Chest,2015,148(4):548A-548B
6邓琳,冯定庆,凌斌.COX2促进卵巢癌细胞迁移及相关机制的研究[J].现代妇产科进展,2016,25(8):561-565
7梁巍,孙文广,姜佳佳,等.TGF-β1诱导人胃腺癌SGC-7901细胞建立EMT模型[J].医学研究杂志,2014,43(6):37-41
8 Zhao R,Gong L,Li L,et al.nm23-H1 is a negative regulator of TGF-β1-dependent induction of epithelial-mesenchymal transition[J].Exp Cell Res,2013,319(5):740-749
9 Kiesslich T,Pichler M,Neureiter D.Epigenetic controlof epithelialmesenchymal-transition in human cancer[J].Mol Clin Oncol,2013,1(1):3-11
10杨燕,邹晓平.上皮间质转化与胃癌的研究进展[J].国际消化病杂志,2014,34(5):334-337
11 Vera M,Barcia E,Negro S,et al.New celecoxib multiparticulate systems to improve glioblastoma treatment[J].Int J Pharm,2014,473(1-2):518-527
12 Zhang Q,Meng X,Zheng G,et al.Antitumor activity of celecoxib,a selective cyclooxygenase-2 inhibitor,in medullary thyroid carcinoma[J].Mol Med Rep,2014,9(2):768-772
13吕丽,李文通,杨灵,等.COX-2表达与乳腺癌EMT发生关系的研究[J].中国妇幼保健,2013,28(28):4728-4730
14李裕明,韩克强,郑璐,等.上皮间质转化标志物E-cadherin和Vimentin在原发性肝癌中的表达及临床意义[J].中华肝脏病杂志,2013,21(4):279-284
15申震,周颖,赵卫东,等.Vimentin及E-Cadherin在子宫颈癌中的表达及其与上皮细胞间质化的相关性[J].安徽医科大学学报,2013,48(5):505-508
16 El-Sayed,Mona,Elazeem A,et al.Expression of epithelial-mesenchymal transition-related markers E-cadherin and vimentin in ovarian serous carcinomas:correlation with transforming growth factor-β1expression and clinicopathological factors[J].Egyptian Journal of Pathology,2016,36(1):1-8
17 Eades G,Yao Y,Yang M,et al.miR-200a regulates SIRT1 expression and epithelial to mesenchymal transition(EMT)-like transformation in mammary epithelial cells.[J].J Biol Chem,2011,286(29):25992-26002
18郝翀.SIRT1在肝细胞癌侵袭转移中的作用机制研究[D].上海:第二军医大学硕士学位论文,2014
19 Byles V,Zhu L,Lovaas JD,et al.SIRT1 induces EMT by cooperating with EMT transcription factors and enhances prostate cancer cell migration and metastasis[J].Oncogene,2012,31(43):4619-4629
20曾薇,朱金峰,于瑞冉,等.miR-34a靶向调控Sirtuin 1表达对食管鳞癌细胞侵袭、转移的影响[J].山东医药,2017,57(24):43-45
21王金凤.白藜芦醇对PC12细胞增殖内稳态影响的研究[D].广州:华南师范大学硕士学位论文,2013