追毒方逆转三阴性乳腺癌MDA-MB-231细胞耐药的作用机制
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摘要
目的基于糖基转移酶β3GnT8探讨追毒方逆转三阴性乳腺癌MDA-MB-231细胞耐药的作用机制。方法通过MTT法观察追毒方对MDA-MB-231耐药细胞增殖的抑制作用,通过转染β3GnT8得到高表达β3GnT8的MDA-MB-231/TAX细胞株,采用Western blot观察糖基转移酶β3GnT8和耐药蛋白BCRP、P-gp的表达,通过凝集素流式细胞术观察转染β3GnT8MDA-MB-231/TAX细胞膜表面β3GnT8的荧光强度。结果追毒方有效成分能明显抑制三阴性乳腺癌MDA-MB-231耐药细胞的增殖,作用72h抑制率达到50.16%,能有效下调β3GnT8和耐药蛋白BCRP、P-gp的表达,并且β3GnT8的表达与BCRP、P-gp呈正相关。结论追毒方可通过下调糖基转移酶β3GnT8,从而降低耐药蛋白BCRP、P-gp的表达,逆转三阴性乳腺癌MDA-MB-231细胞耐药性。
OBJECTIVE Based on glycosyltransferaseβ3 GnT8,to investigate the mechanism of reverse effect on triple negative breast cancer MDA-MB-231 drug-resistance cells by Zhuidu Fang.METHODS The inhibition of MDA-MB-31 drug-resistance cells proliferation by Zhuidu Fang was detected by MTT assay.By interference and transfection ofβ3 GnT8,high and low expressedβ3 GnT8 MDA-MB-231/TAX cell lines were obtained.Western blot was used to detect the protein expression ofβ3 GnT8,and drug resistance protein BCRP and P-gp.Lectin flow cytometry was used to detectβ3 GnT8 fluorescence intensity onβ3 GnT8 transfected MDA-MB-231/TAX cells.RESULTS Zhuidu Fang effectively inhibited the proliferation of MDA-MB-231/TAX cells,and the inhibitory rate was 50.16%after 72 h.Zhuidu Fang significantly downregulated the protein expression ofβ3 GnT8,BCRP and P-gp.The protein expression ofβ3 GnT8 showed a positive correlation with BCRP and P-gp.CONCLUSION Zhuidu Fang can reverse the drug-resistance of MDA-MB-231 cell lines,which is associated with the downregulation of the expression ofβ3 GnT8,BCRP and P-gp.
OBJECTIVE Based on glycosyltransferaseβ3 GnT8,to investigate the mechanism of reverse effect on triple negative breast cancer MDA-MB-231 drug-resistance cells by Zhuidu Fang.METHODS The inhibition of MDA-MB-31 drug-resistance cells proliferation by Zhuidu Fang was detected by MTT assay.By interference and transfection ofβ3 GnT8,high and low expressedβ3 GnT8 MDA-MB-231/TAX cell lines were obtained.Western blot was used to detect the protein expression ofβ3 GnT8,and drug resistance protein BCRP and P-gp.Lectin flow cytometry was used to detectβ3 GnT8 fluorescence intensity onβ3 GnT8 transfected MDA-MB-231/TAX cells.RESULTS Zhuidu Fang effectively inhibited the proliferation of MDA-MB-231/TAX cells,and the inhibitory rate was 50.16%after 72 h.Zhuidu Fang significantly downregulated the protein expression ofβ3 GnT8,BCRP and P-gp.The protein expression ofβ3 GnT8 showed a positive correlation with BCRP and P-gp.CONCLUSION Zhuidu Fang can reverse the drug-resistance of MDA-MB-231 cell lines,which is associated with the downregulation of the expression ofβ3 GnT8,BCRP and P-gp.
引文
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[2]王晓稼.2015年美国ASCO年会三阴性乳腺癌研究进展[J].中国肿瘤外科杂志,2015,7(3):141-145.
[3]刘敏,王明武.解毒破瘀法抑制三阴性乳腺癌复发与转移的临床研究[J].南京中医药大学学报,2016,32(2):111-113.
[4]LIU CL,QIU H,LIN DD,et al.c-Jun-dependentβ3GnT8promotes tumorigenesis and metastasis of hepatocellular carcinoma by inducing CD147glycosylation and altering N-glycan patterns[J].Oncotarget,2018(1):1-13.
[5]QIU H,XU X,LIU M,et al.RNA interference mediated silencing of ppGalNAc T1and ppGalNAc T2inhibits invasion and increases chemosensitivitypotentially by reducing terminalα2,3sialylation and MMP14expression in triple negative breast cancer cells[J].Mol Med Rep,2017(15):3724-3734.
[6]WANG Y,LU JJ,HE L,et al.Triptolide(TPL)inhibits global transcription by inducing proteasome-dependent degradation of RNA polymerase II(Pol II)[J].PLoS ONE,2011,6:e23993.
[7]PAN L,CHAI H,KINGHORN AD.The continuing search for antitumor agents from higher plants[J].Phytochem Lett,2010,3(1):1-8.
[8]CAREY N,LA THANGUE NB.Histone deacetylase inhibitors:gathering pace[J].Curr Opin Pharmacol,2006,6(4):369-375.
[9]GUO JM,CHEN HL,Wang GM,et al.Expression of UDPGalNAc:polypeptide N-acetylgalactosaminyltransferase-12 in gastric and colonic cancer cell lines and in human colorectal cancer[J].Oncology,2004,67:271-276.
[10]HUANG C,ZHOU J,WU S,et al.Cloning and tissue distribution of the human B3GALT7gene,a member of the beta1,3-Glycosyltransferase family[J].Glycoconj J,2004,21:267-273.
[11]LIU CL,QIU H,YU MY,et al.c-Jun-mediatedβ-1,3-Nacetylglucosaminyltransferase 8expression:A novel mechanism regulating the invasion and metastasis of colorectal carcinoma cells[J].Oncol Lett,2017,14:3722-3728.
[12]刘敏.中药解毒成分复方对乳腺癌作用和机理的实验研究[D].南京:南京中医药大学,2014.