蒲公英提取物抑制人胃癌细胞侵袭迁移能力的实验研究
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摘要
目的通过研究蒲公英提取物(ETM)对人胃低分化腺癌SGC-7901细胞增殖、迁移和侵袭的影响,探讨ETM抑制胃癌细胞侵袭转移的作用和可能机制。方法体外培养胃癌SGC-7901细胞,以不同浓度的ETM进行处理,CCK-8法检测细胞增殖活性;细胞划痕实验观察细胞迁移能力;Transwell小室实验检测细胞侵袭能力;实时荧光定量聚合酶链式反应(qRT-PCR)法检测丝氨酸/苏氨酸蛋白激酶的一种:单丝氨酸蛋白激酶(LIMK1),以及基质金属蛋白酶9(MMP9)、粘着斑激酶(FAK)mRNAs表达;Western blot法检测LIMK1、MMP9、FAK蛋白表达。结果 ETM对SGC-7901细胞增殖活力具有明显的抑制作用;ETM浓度达50μmol/L以上时,能抑制SGC-7901细胞侵袭及迁移能力,显著下调LIMK1、MMP9、FAK mRNAs及其蛋白表达,且抑制和下调作用均呈浓度和时间依赖性(P均<0.01)。结论 ETM能抑制SGC-7901细胞的侵袭和迁移能力,其作用机制与下调LIMK1、MMP9、FAK的表达有关。
Objective To investigate the effect of extract from taraxacum monogon(ETM)on proliferation,migration and invasion of human poorly differentiated gastric adenocarcinoma SGC-7901 cells and its possible mechanism.Methods The SGC-7901 cells were cultured in vitro and were treated with different concentration of ETM.The proliferation activity of tumor cells was detected by Cell Counting Kit-8(CCK-8) test.The migaratory ability of tumor cells was detected by wound healing test.The invasive ability of tumor cells was detected by Transwell test.The expression of single serine protein kinase 1(LIM domain kinase 1,LIMK1),a serine/threonine-protein kinase,matrix metalloproteinases 9(MMP9) and focal adhesion kinase(FAK) mRNAs were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The expression of LIMK1,MMP9 and FAK proteins were detected by Western-blot test.Results ETM could significantly inhibit proliferation,invasion and migration abilities of SGC-7901 cells and down-regulate the expression of LIMK1,MMP9 and FAK mRNAs and proteins.Both inhibition and down-regulation were concentration-and time-dependent(all P<0.01).Conclusion ETM can inhibit invasion and migration abilities of SGC-7901 cells,and the mechanism may be related to down-regulating the expression of LIMK1,MMP9 and FAK.
Objective To investigate the effect of extract from taraxacum monogon(ETM)on proliferation,migration and invasion of human poorly differentiated gastric adenocarcinoma SGC-7901 cells and its possible mechanism.Methods The SGC-7901 cells were cultured in vitro and were treated with different concentration of ETM.The proliferation activity of tumor cells was detected by Cell Counting Kit-8(CCK-8) test.The migaratory ability of tumor cells was detected by wound healing test.The invasive ability of tumor cells was detected by Transwell test.The expression of single serine protein kinase 1(LIM domain kinase 1,LIMK1),a serine/threonine-protein kinase,matrix metalloproteinases 9(MMP9) and focal adhesion kinase(FAK) mRNAs were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The expression of LIMK1,MMP9 and FAK proteins were detected by Western-blot test.Results ETM could significantly inhibit proliferation,invasion and migration abilities of SGC-7901 cells and down-regulate the expression of LIMK1,MMP9 and FAK mRNAs and proteins.Both inhibition and down-regulation were concentration-and time-dependent(all P<0.01).Conclusion ETM can inhibit invasion and migration abilities of SGC-7901 cells,and the mechanism may be related to down-regulating the expression of LIMK1,MMP9 and FAK.
引文
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[13] 李春雨,王琪,申珅,等.冬凌草甲素对人肝癌MHCC-97H细胞系侵袭和迁移的作用[J].中国病理生理杂志,2017,33(8):1423-1427.
[14] 刘鑫慧,杨清.LIMK的研究进展[J].现代肿瘤医学,2018,26(1):137-141.
[15] Lagoutte E,Villeneuve C,Lafanechère L,et al.LIMK regulates tumor-cell invasion and matrix degradation through tyrosine phosphorylation of MT1-MMP[J].Sci Rep,2016,6:24925.
[16] Shanthi E,Krishna MH,Arunesh GM,et al.Focal adhesion kinase inhibitors in the treatment of metastatic cancer:a patent review[J].Expert Opin Ther Pat,2014,24(10):1077-1100.
[17] Kanteti R,Mirzapoiazova T,Riehm JJ,et al.Focal Adhesion Kinase a Potential Therapeutic Target for Pancreatic Cancer and Malignant Pleural Mesothelioma[J].Cancer Biol Ther 2018,19(4):316-327.
[18] Tomar S,Plotnik JP,Haley J,et al.ETS1 induction by the microenvironment promotes ovarian cancer metastasis through focal adhesion kinase[J].Cancer Lett,2018,414:190-204.
[19] Yousef EM,Tahir MR,St-Pierre Y,et al.MMP-9 expression varies according to molecular subtypes of breast cancer[J].BMC Cancer,2014,14:609.
[2] 罗玉政,陈波,李红樱.沉默IGF1R表达对胃癌细胞侵袭与迁移能力的影响及机制[J].中国临床研究,2018,31(2):163-166.
[3] 黄昊,杨星九,高苒.胃癌多药耐药机制及研究进展[J].中国医学科学院学报,2016,38(6):739-745.
[4] 郭君宾,叶海虹,陈剑峰.蒲公英提取物抗人肝癌细胞HepG2增殖的作用及机制研究[J].中药材,2015,38(10):2129-2133.
[5] 齐绪林,高鹏飞,乔田奎,等.中药蒲公英抗肿瘤作用研究进展[J].中国肿瘤,2015,24(1):53-56.
[6] 蒙旭.乙酰蒲公英萜醇影响HepG2细胞增殖、凋亡及迁移的实验研究[D].昆明:昆明医科大学,2016.
[7] 呼永华.蒲公英的抗癌机理研究[J].西部中医药,2018,31(1):132-134.
[8] 石国慧,陈远才,王旭德,等.药食同源品蒲公英抗肿瘤活性及其作用机制的研究进展[J].沈阳药科大学学报,2017,34(9):858-862.
[9] 农云宏,白浪,唐红.tenascin-C促进肿瘤转移的分子机制研究进展[J].生物医学工程学杂志,2015,32(1):240-244.
[10] Davila M,Frost AR,Grizzle WE,et al.LIM kinase 1 is essential for the invasive growth of prostate epithelial cells:implications in prostate cancer[J].J Biol Chem,2003,278(38):36868-36875.
[11] 吴南芳,张琍.LIMK1/Cofilin信号通路与肿瘤的研究进展[J].微生物学免疫学进展,2016,44(1):72-75.
[12] 苏坚,潘志兵,史玲,等.LIMK1在结肠癌中表达与沉默对人结肠癌细胞迁移与侵袭的影响[J].中南医学科学杂志,2014,42(2):109-115.
[13] 李春雨,王琪,申珅,等.冬凌草甲素对人肝癌MHCC-97H细胞系侵袭和迁移的作用[J].中国病理生理杂志,2017,33(8):1423-1427.
[14] 刘鑫慧,杨清.LIMK的研究进展[J].现代肿瘤医学,2018,26(1):137-141.
[15] Lagoutte E,Villeneuve C,Lafanechère L,et al.LIMK regulates tumor-cell invasion and matrix degradation through tyrosine phosphorylation of MT1-MMP[J].Sci Rep,2016,6:24925.
[16] Shanthi E,Krishna MH,Arunesh GM,et al.Focal adhesion kinase inhibitors in the treatment of metastatic cancer:a patent review[J].Expert Opin Ther Pat,2014,24(10):1077-1100.
[17] Kanteti R,Mirzapoiazova T,Riehm JJ,et al.Focal Adhesion Kinase a Potential Therapeutic Target for Pancreatic Cancer and Malignant Pleural Mesothelioma[J].Cancer Biol Ther 2018,19(4):316-327.
[18] Tomar S,Plotnik JP,Haley J,et al.ETS1 induction by the microenvironment promotes ovarian cancer metastasis through focal adhesion kinase[J].Cancer Lett,2018,414:190-204.
[19] Yousef EM,Tahir MR,St-Pierre Y,et al.MMP-9 expression varies according to molecular subtypes of breast cancer[J].BMC Cancer,2014,14:609.