加味芍药甘草汤对子宫腺肌症细胞增殖和迁移能力的影响
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摘要
目的:研究加味芍药甘草汤对人子宫腺肌细胞增殖和迁移能力的影响。方法:取人子宫腺肌细胞进行细胞培养,随机分为中药实验组(加味芍药甘草汤水提液)、西药对照组(米非司酮片)和空白对照组3组,分别予对应的药物处理24 h和48 h后,通过四唑盐比色法(MTT法)检测药物对细胞增殖的影响;细胞划痕实验检测药物对细胞迁移能力的影响;Western blot检测药物对细胞中EGFR、p-EGFR、STAT3、p-STAT3、COX-2蛋白表达的影响。结果:与西药对照组和空白对照组相比,中药实验组的人子宫腺肌细胞增殖受到显著抑制,划痕实验后细胞愈合能力减弱,并且EGFR、p-EGFR、STAT3、p-STAT3、COX-2蛋白的表达量下降(P<0.05)。结论:加味芍药甘草汤能在一定程度上抑制人子宫腺肌细胞的增殖、降低细胞迁移率,其作用机制可能是通过调控EGFR/STAT3/COX-2信号通路而实现的。
Objective: To explore the effects of Jiawei Shaoyao Gancao Decoction on the proliferation and migration of Adenomyosis cells. Methods: The human adenomyosis cells were cultured in vitro and divided into three groups as Chinese medicine group( Jiawei Shaoyao Gancao Decocotion),the positive control group( Mifepristone) and the blank control group in random with different interventions respectively. After 24 h and 48 h,cells proliferation was detected by MTT assay,and migration was investigated by wound-healing test. Western blot was used to determine the expression levels of EGFR,p-EGFR,STAT3,p-STAT3 and COX-2 in each group. Results: Compared with the positive control group and blank control group,the proliferation and migration of cells in Chinese medicine group was inhibited obviously( P < 0. 05). And Western blot test showed that there was down-regulated expressions of EGFR,p-EGFR,STAT3,p-STAT3 and COX-2 in Chinese medicine group in contrast to those of the other groups( P < 0. 05). Conclusion: Jiawei Shaoyao Gancao Decoction may prohibit the proliferation and migration of the human adenomyosis cells in vitro by regulating EGFR/STAT3/COX-2 signal pathway.
Objective: To explore the effects of Jiawei Shaoyao Gancao Decoction on the proliferation and migration of Adenomyosis cells. Methods: The human adenomyosis cells were cultured in vitro and divided into three groups as Chinese medicine group( Jiawei Shaoyao Gancao Decocotion),the positive control group( Mifepristone) and the blank control group in random with different interventions respectively. After 24 h and 48 h,cells proliferation was detected by MTT assay,and migration was investigated by wound-healing test. Western blot was used to determine the expression levels of EGFR,p-EGFR,STAT3,p-STAT3 and COX-2 in each group. Results: Compared with the positive control group and blank control group,the proliferation and migration of cells in Chinese medicine group was inhibited obviously( P < 0. 05). And Western blot test showed that there was down-regulated expressions of EGFR,p-EGFR,STAT3,p-STAT3 and COX-2 in Chinese medicine group in contrast to those of the other groups( P < 0. 05). Conclusion: Jiawei Shaoyao Gancao Decoction may prohibit the proliferation and migration of the human adenomyosis cells in vitro by regulating EGFR/STAT3/COX-2 signal pathway.
引文
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[25]薛刚,邹玺,周锦勇,等.竹节香附素A对人胃癌细胞株BGC-823细胞周期及STAT3信号通路的影响[J].南京中医药大学学报,2017,33(1):78-81.
[26]Feng Z,Zhang C,Kang H J,et al.Regulation of female reproduction by p53 and its family members[J].Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology,2011,25(7):2464-2468.
[27]Yu H,Yue X,Zhao Y,et al.LIF negatively regulates tumour-suppressor p53 through Stat3/ID1/MDM2 in colorectal cancers[J].Nat Commun,2014(5):5218.
[28]Yahata Y,Shirakata Y,Tokumaru S,et al.Nuclear translocation of phosphorylated STAT3 is essential for vascular endothelial growth factor-induced human dermal microvascular endothelial cell migration and tube formation[J].J Biol Chem,2003,278(41):40026-40031.
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[30]曾玉燕,关永格,李坤寅.雌激素及其受体、芳香化酶在子宫腺肌病中的作用[J].南方医科大学学报,2017,37(3):383-387.
[31]Wang S W,Sun Y M.The IL-6/JAK/STAT3 pathway:potential therapeutic strategies in treating colorectal cancer(Review)[J].Int J Oncol,2014,44(4):1032-1040.
[2]Struble J,Reid S,Bedaiwy M A.Adenomyosis:A Clinical Review of a Challenging Gynecologic Condition[J].Journal of Minimally Invasive Gynecology,2015,23(2):164-185.
[3]T Harada,YM Khine,A Kaponis,et al.The Impact of Adenomyosis on Women’s Fertility[J].Obstetrical&Gynecological Survey,2016,71(9):557-568.
[4]王帅.加味芍药甘草汤对子宫腺肌病MAPK/ERK信号通路干预机制[D].广州:广州中医药大学,2015.
[5]曾玉燕.子宫腺肌病ER及mi R-21介导的Ras/Raf通路作用机制及加味芍药甘草汤对其的干预[D].广州:广州中医药大学,2015.
[6]Garcia L,Isaacson K.Adenomyosis:Review of the Literature[J].Reference Librarian,2011,18(4):428-437.
[7]Gordts S,Campo R,Brosens I.Hysteroscopic diagnosis and excision of myometrial cystic adenomyosis[J].Gynecological Surgery,2014,11(4):273-278.
[8]Schindler A E.Non-contraceptive benefits of oral hormonal contraceptives[J].Minerva Ginecologica,2014,11(1):319-329.
[9]C Pafumi,M Farina,G Pernicone,et al.Adenomyosis and uterus rupture during labor[J].Chinese medical journal,2017,64(4):244-246.
[10]L P,C D,C D.Update on the surgical management of adenomyosis[J].Applied Optics,2012,24(4):3732-3737.
[11]Baselga J.A review of EGFR targeted therapy[J].Clinical Advances in Hematology&Oncology H&O,2003,1(4):218-219.
[12]Vazquez-MA,Cuf S,Oliveras FC,et al.IGF-1R/epithelial-to-mesenchymal transition(EMT)crosswalk suppresses the erlotinib-sensitizing effect of EGFR exon 19 deletion mutations[J].Sci Rep,2013(3):2560.
[13]F Steiner,C Hauserkronberger,G Rendl,et al.Expression of Tenascin C,EGFR,E-Cadherin,and TTF-1 in Medullary Thyroid Carcinoma and the Correlation with RET Mutation Status[J].International Journal of Molecular Sciences,2016,17(7):1093.
[14]Nagathihalli NS,Merchant NB.Src-mediated regulation of E-cadherin and EMT in pancreatic cancer[J].Biosci,2012,17:2059-2069.
[15]Mohamed Y.The value of E-cadherin and EGFR expression in ovarian serous tumors[J].Journal of American Science,2016,12(2):53-61.
[16]郭海雁,张颖,黄春建,等.子宫腺肌病组织中ER、PR和EGFR的表达及其临床意义[J]中国医药科学,2012,2(19):20-22.
[17]孙栋勋,蔡志毅.EGFR/PI3K/Akt细胞信号传导通路与肿瘤[J].检验医学,2014,29(7):768-773.
[18]Cheng GZ,Park S,Shu S,et al.Advances of AKT pathway in human oncogenesis and as a target for anti-cancer drug discovery[J].Curr Cancer Drug Targets,2008,8(1):2-6.
[19]Okazaki H,Tokumaru S,Hanakawa Y,et al.Nuclear translocation of phosphorylated STAT3 regulates VEGF-A-induced lymphatic endothelial cell migration and tube formation[J].Biochem Biophys Res Commun,2011,412(3):441-445.
[20]Weimar C H,Macklon N S,Post U E,et al.The motile and invasive capacity of human endometrial stromal cells:implications for normal and impaired reproductive function[J].Hum Reprod Update,2013,19(5):542-557.
[21]王静,杨阳,杨宁,等.磷酸化信号转导和转录激活因子3(Y705)在子宫腺肌症中的表达及其临床意义[J].解剖学报,2016(2):261-267.
[22]张红霞,吴广胜.β-arrestin1激活STAT3信号通路影响白血病细胞K562体外迁移侵袭能力[J].中国免疫学杂志,2016,12(32):1773-1776.
[23]胡文龙,殷嫦嫦,吴平平,等.脐带间充质干细胞通过IL-6/STAT3信号通路促进骨肉瘤细胞增殖和迁移[J].中国细胞生物学学报,2016,38(1):24-31.
[24]来伟,施景龙,林显敢,等.IL-6/p-AKT/p-STAT3信号通路在PRL-3促进结肠癌细胞增殖、迁移中的机制研究[J].岭南现代临床外科,2016,16(5):517-520.
[25]薛刚,邹玺,周锦勇,等.竹节香附素A对人胃癌细胞株BGC-823细胞周期及STAT3信号通路的影响[J].南京中医药大学学报,2017,33(1):78-81.
[26]Feng Z,Zhang C,Kang H J,et al.Regulation of female reproduction by p53 and its family members[J].Faseb Journal Official Publication of the Federation of American Societies for Experimental Biology,2011,25(7):2464-2468.
[27]Yu H,Yue X,Zhao Y,et al.LIF negatively regulates tumour-suppressor p53 through Stat3/ID1/MDM2 in colorectal cancers[J].Nat Commun,2014(5):5218.
[28]Yahata Y,Shirakata Y,Tokumaru S,et al.Nuclear translocation of phosphorylated STAT3 is essential for vascular endothelial growth factor-induced human dermal microvascular endothelial cell migration and tube formation[J].J Biol Chem,2003,278(41):40026-40031.
[29]Wang X,Crowe P J,Goldstein D,et al.STAT3 inhibition,a novel approach to enhancing targeted therapy in human cancers(review)[J].Int J Oncol,2012,41(4):1181-1191.
[30]曾玉燕,关永格,李坤寅.雌激素及其受体、芳香化酶在子宫腺肌病中的作用[J].南方医科大学学报,2017,37(3):383-387.
[31]Wang S W,Sun Y M.The IL-6/JAK/STAT3 pathway:potential therapeutic strategies in treating colorectal cancer(Review)[J].Int J Oncol,2014,44(4):1032-1040.