王宏健,刘晓红,董宇华,郭阳,王旭东
WANG Hongjian,LIU Xiaohong,DONG Yuhua,GUO Yang,WANG Xudong

• 1:贵州医科大学基础医学院生理学教研室

摘要(Abstract)：

目的:观察蛋白激酶A(PKA)的抑制剂H89在雌激素(E2)依赖人乳腺癌细胞(MCF-7)纤连蛋白(fibronectin,FN)表达及其与CANP2的关系。方法:分别用E2和PKA抑制剂H89刺激对数生长期MCF-7细胞和CANP2-shRNA转染MCF-7模型细胞,采用伤口愈合实验检测细胞迁移率、Western-blot法测FN蛋白的表达。结果:与对照组比较,H89预处理后,E2诱导的MCF-7细胞迁移率可明显增加,E2与H89联用可使FN蛋白表达明显上调,差异有统计学意义(P<0.05);H89和E2+H89组的可促使CANP2-shRNA转染MCF-7细胞的迁移率明显降低,并抑制CANP2-shRNA转染MCF-7细胞中FN蛋白表达,差异有统计学意义(P<0.01)。结论:PKA抑制剂能明显增强E2诱导的乳腺癌细胞迁移和FN蛋白表达,而该效应可被CANP2基因沉默所逆转。
Objective:To observe the impact of H89 on the cell migration and the protein expression of fibronectin(FN) in MCF-7 breast cancer cells and its relationship with calpain2.Methods:Estrogen(E2) and protein kinase A(PKA) inhibitor H89 were used to stimulate MCF-7 cells and CANP2-shRNA transfected model cells in logarithmic growth phase respectively.Cell migration and FN protein expression were measured by wound healing test and Western-blot method.Results:Compared with the control group,the cell migration rate induced by E2 increased significantly after H89 pretreatment(P<0.05),and the expression of FN protein increased significantly after E2 combined with H89(P<0.01).After gene silencing of calcium-activated neutral protease 2(CANP2),the cell migration of H89 group and E2 + H89 group decreased significantly(P<0.01),and the up-regulation of FN protein in H89 group and E2 + H89 group was significantly inhibited(P<0.01 compared with NC group).Conclusion:PKA inhibitors can significantly enhance E2-induced cell migration and FN protein expression in breast cancer cells,which can be reversed by CANP2 gene silencing.

关键词(KeyWords)： 乳腺肿瘤;雌激素类;蛋白酶抑制剂;纤连蛋白类;细胞迁移;钙蛋白酶2
breast neoplasms;estrogen;protease inhibitors;fibronectin;cell migration;calpain 2

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅-贵州医科大学科技合作计划项目[黔科合LH(2015)7317];; 国家自然科学基金项目(31360252,31660345)

作者(Author): 王宏健,刘晓红,董宇华,郭阳,王旭东
WANG Hongjian,LIU Xiaohong,DONG Yuhua,GUO Yang,WANG Xudong

DOI: 10.19367/j.cnki.1000-2707.2019.07.010

参考文献(References)：

• [1]AKRAM M,IQBAL M,DANIYAL M,et al. Awareness and current knowledge of breast cancer[J]. Biol Res,2017,50(1):33.
• [2]PUPO M,PISANO A,LAPPANO R,et al. Bisphenol A induces gene expression changes and proliferative effects through GPER in breast cancer cells and cancer-associated fibroblasts[J]. Environ Health Perspect,2012,120(8):1177-1182.
• [3]SAMPSON J N,FALK R T,SCHAIRER C et al. Association of estrogen metabolism with breast cancer risk in different cohorts of postmenopausal women[J]. Cancer Res,2017,77(4):918-925.
• [4]YU C L,SE L K,YOUNG R P,et al. Parthenolide promotes apoptotic cell death and inhibits the migration and invasion of SW620 cells[J]. Intest Res,2017,15(2):174-181.
• [5]YU L L,YUAN D Z,ZHANG S M,et al. Progress of nongenomic action of estrogen and its impact on female reproduction[J]. Sheng Li Xue Bao,2016,68(4):547-556.
• [6]LAU K M,MA F M,XIA J T,et al. Activation of GPR30stimulates GTP-binding of Gαi1 protein to sustain activation of Erk1/2 in inhibition of prostate cancer cell growth and modulates metastatic properties[J]. Experimental Cell Research,2017,350(1):199-209.
• [7]YU X,STALLONE J N,HEAPS C L,et al. The activation of G protein-coupled estrogen receptor induces relaxation via c AMP as well as potentiates contraction via EGFR transactivation n porcine coronary arteries[J]. PLo S One,2018,13(1):e0191418.
• [8]WANG J P,HIELSCHER A. Fibronectin:How its aberrant expression in tumors may improve therapeutic targeting[J]. Journal of Cancer,2017,8(4):674-682.
• [9]JEONG Y,YOU D,KANG H G,et al. Berberine suppresses fibronectin expression through inhibition of c-Jun phosphorylation in breast cancer cells[J]. J Breast Cancer,2018,21(1):21-27.
• [10]WANG X D,ROSALES J L,MAGLIOCCO A,et al.Cyclin E in breast tumors is cleaved into its low molecular weight forms by calpain[J]. Oncogene,2003,22(5):769-774.
• [11]RIDLEY A J,SCHWARTZ M A,BURRIDGE K,et al.cell migration:integrating signals from front to back[J].Science,2003,302(5651):1704-1709.
• [12]VELLOSO F J,BIANCO A F,FARIAS J O,et al. The crossroads of breast cancer progression:insights into the modulation of major signaling pathways[J]. Onco Targets and Therapy,2017,10:5491-5524.
• [13]任伟宏.蛋白激酶与细胞凋亡信号及凋亡抑制信号的转导[J].国外医学:临床生物化学与检验学分册,2004,25(1):79-82.
• [14]FILARDO E J,QUINN J A,FRACKELTON A R J,et al. Estrogen action via the G protein-coupled receptor,GPR30:stimulation of adenylyl cyclase and c AMP-mediated attenuation of the epidermal growth factor receptorto-MAPK signaling axis[J]. Mol Endocrinol,2002,16(1):70-84.
• [15]CLARYSSE L,GUGUINOU M,POTIER-CARTEREAU M,et al. c AMP-PKA inhibition of SK3 channel reduced both Ca2+entry and cancer cell migration by regulation of SK3-Orai1 complex[J]. Pflugers Arch,2014,466(10):1921-1932.
• [16]LEE J W,LEE J,MOON E Y. He La human cervical cancer cell migration is inhibited by treatment with dibutyryl-cAMP[J]. Anticancer Res,2014,34(7):3447-3455.
• [17]KENNY H A,CHIANG C Y,WHITE E A,et al. Mesothelial cells promote early ovarian cancer metastasis through fibronectin secretion[J]. The Journal of Clinical Investigation,2014,124(10):4614-4628
• [18]EKE I,STORCH K,KRAUSE M,et al. Cetuximab attenuates its cytotoxic and radiosensitizing potential by inducing fibronectin biosynthesis[J]. Cancer Research,2013,73(19):5869-5879
• [19]于庆龙,张莹,王宏健,等. 17β-雌二醇对乳腺癌细胞迁移的影响及CANP-FN通路的介导作用[J].中国药理学通报,2017,33(10):1371-1376.
• [20]WU Q,DHIR R,WELLS A. Altered CXCR3 isoform expression regulates prostate cancer cell migration and invasion[J]. Mol Cancer,2012,11(1):3.

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