全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
-  2017 

Hh信号通路抑制剂Cyclopamine对Hela细胞增殖、侵袭迁移的影响 Effect of Cyclopamine,an Hh Signal Pathway Inhibitor,on the Proliferation,Invasion and Metastasis of Hela Cells

Keywords: Hh信号通路,Hela细胞,增殖,侵袭迁移,Gli1,FoxM1,EMT

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的:研究Hh信号通路抑制剂Cyclopamine作用Hela细胞后Gli1、FoxM1、上皮间质转化(EMT)过程相关标记物表达的改变及对其增殖、侵袭迁移能力的影响,探究Gli1、FoxM1、EMT过程的可能作用关系,以期为宫颈癌的分子靶向治疗提供多个靶点。方法:采用实时定量RT-PCR、Western Blot技术检测Hela、Siha细胞系Hh信号通路中Shh、Ptch1、Smo的表达情况,选择Hh信号通路表达较高的Hela细胞系作为后续实验细胞,利用四甲基偶氮唑蓝比色法(MTT法)检测不同浓度Cyclopamine作用Hela细胞后其增殖能力的改变;采用Transwell小室试验检测Hela细胞侵袭、迁移能力的改变,并采用实时定量RT-PCR、Western Blot技术检测Gli1、FoxM1、EMT过程标记物的表达情况。结果:(1)Hela细胞中Hh信号通路成分(Shh、Ptch1、Smo)在mRNA水平显著高于Siha细胞(P<0.01),且相应蛋白的表达强度也高于Siha细胞;(2)Cyclopamine对Hela细胞增殖的抑制作用具有时间、剂量依赖性;(3)Cyclopamine作用Hela细胞48h后,细胞侵袭试验显示,实验组侵袭细胞数较对照组明显降低(P<0.01)。细胞迁移试验显示,实验组细胞迁移细胞数较对照组明显降低(P<0.01);(4)Cyclopamine作用Hela细胞48h后,实验组Gli1、FoxM1、Vimentin在mRNA水平的表达显著低于对照组(P<0.05),且相应蛋白的表达强度也低于对照组;E-cadherin在实验组中mRNA层面的表达显著高于对照组(P<0.05),且相应蛋白的表达强度也高于对照组。结论:抑制Hh信号通路可显著降低Hela细胞系增殖、侵袭迁移能力,FoxM1很可能是Hh信号通路末端转录因子Gli1的下游靶基因,且其对宫颈癌细胞系增殖、侵袭迁移的作用很可能是通过EMT实现的

References

[1]  Siegel R,Jiemin Ma PhD,Zhaohui Zou MS,et al.Cancer statistics,2014[J].CA:A Cancer Journal for Clinicians,2014,64(1):9-29.
[2]  Bernard HU,Burk RD,Chen Z,et al.Classification of papillomaviruses(PVs)based on 189 PV types and proposal of taxonomic amendments[J].Virology,2010,401(1):70-79.
[3]  Fischer KR,Durrans A,Lee S,et al.Epithelial-tomesenchymal transition is not required for lung metastasis but contributes to chemoresistance[J].Nature,2015,527(7 579):472-476.
[4]  Xiaofeng Z,Carstens JL,Jiha K,et al.Epithelial-tomesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer[J].Nature,2015,527(7 579):525-530.
[5]  Solanas G,Benitah SA.Regenerating the skin:a task for the heterogeneous stem cell pool and surrounding niche[J].Nature Reviews Molecular Cell Biology,2013,14(11):737-748.
[6]  Kugler MC,Joyner AL,Loomis CA,et al.Sonic hedgehog signaling in the lung.From development to disease[J].American Journal of Respiratory Cell&Molecular Biology,2015,52(1):1-13.
[7]  Li H,Li J,Feng L.Hedgehog signaling pathway as a therapeutic target for ovarian cancer[J].Cancer Epidemiology,2016,40:152-157.
[8]  Irvine DA,Mhairi C.Targeting hedgehog in hematologic malignancy[J].Blood,2012,119(119):2 196-2 204.
[9]  Teh MT,Wong ST,Neill GW,et al.FOXM1is a downstream target of Gli1in basal cell carcinomas[J].Cancer Research,2002,62(16):4 773-4 780.
[10]  Ling Y,Jing H,Huang S,et al.Activation of hedgehog signaling is not a frequent event in ovarian cancers[J].Molecular Cancer,2009,8(1):1-5.
[11]  Samarzija I,Beard P.Hedgehog pathway regulators influence cervical cancer cell proliferation,survival and migration[J].Biochemical&Biophysical Research Communications,2012,425(1):64-69.
[12]  Yan R,Peng X,Yuan X,et al.Suppression of growth and migration by blocking the Hedgehog signaling pathway in gastric cancer cells[J].Cellular Oncology,2013,36(5):421-435.
[13]  陈丹,陈红,董良波,等.Gli1与FoxM1在子宫颈癌组织和细胞中的表达及意义[J].武汉大学学报:医学版,2016,37(3):430-435.Chen D,Chen H,Dong LB,et al.Significance and Expression of Gli1and FoxM1in cervical cancer cells in vitro and in vivo[J].Medical Journal of Wuhan University,2016,37(3):430-435.
[14]  Thiery JP,Acloque H,Ruby Y,et al.Epithelial-Mesenchymal Transitions in Development and Disease[J].Journal of Oral&Maxillofacial Pathology,2009,139(5):871-890.
[15]  Kalluri R,Weinberg RA.The basics of epithelial-mesenchymal transition[J].Journal of Clinical Investigation,2009,119(6):1 420-1 428.
[16]  肖长纪,陈红,陈慧君,等.下调FoxM1基因表达对子宫颈癌细胞增殖能力的影响[J].中华妇产科杂志,2013,48(7):542-544.Xiao CJ,Chen H,Chen HJ,et al.The effection on the proliferation of cervical cancer cells after Down-regulation of FoxM1gene expression[J].Chinese Journal of Clinical Obstetrics and Gynecology,2013,48(7):542-544.
[17]  Zhang J,Tian XJ,Xing J.Signal transduction pathways of EMT induced by TGF-β,SHH,and WNT and their crosstalks[J].Journal of Clinical Medicine,2016,5(4):1-41.
[18]  Yu C,Chen L,Yie L,et al.Targeting FoxM1inhibits proliferation,invasion and migration of nasopharyngeal carcinoma through the epithelialto-mesenchymal transition pathway[J].Oncology Reports,2015,33(5):2 402-2 410.
[19]  Varjosalo M,Taipale J.Hedgehog:functions and mechanisms[J].Genes&Development,2008,22(18):2 454-2 472.
[20]  Ihrie R,Shah J,Harwell C,et al.Persistent sonic hedgehog signaling in adult brain determines neural stem cell positional identity[J].Neuron,2011,71(2):250-262.
[21]  Su JL,Do IG,Lee J,et al.Gastric cancer(GC)patients with hedgehog pathway activation:PTCH1and GLI2as independent prognostic factors[J].Targeted Oncology,2013,8(4):47-58.
[22]  Horowitz JC,Thannickal VJ.Epithelial-mesenchymal interactions in pulmonary fibrosis[J].Annual Review of Physiology,2010,73(1):413-435.
[23]  Mori Y,Okumura T,Tsunoda S,et al.Gli-1expression is associated with lymph node metastasis and tumor progression in esophageal squamous cell carcinoma[J].Oncology,2006,70(5):378-389.
[24]  Song L,Wang W,Liu D,et al.Targeting of sonic hedgehog-Gli signaling:A potential therapeutic target for patients with breast cancer[J].Oncology Letters,2016,12(2):1 027-1 033.
[25]  Jemal A,Bray F,Center MM,et al.Global cancer statistics[J].CA Cancer J Clin,2011,61:69-90.
[26]  Chen W,Zheng R,Zeng H,et al.Annual report on status of cancer in China,2011[J].Chinese Journal of Cancer Research:English Edition,2015,27(1):2-12.
[27]  Giuliano AR,Harris R,Sedjo RL,et al.Incidence,prevalence,and clearance of type-specific human papillomavirus infections:the young women’s health study[J].Journal of Infectious Diseases,2002,186(4):462-9.
[28]  Kim Y,Kim MO,Jin SS,et al.Hedgehog Signaling Between Cancer Cells and Hepatic Stellate Cells in Promoting Cholangiocarcinoma[J].Annals of Surgical Oncology,2014,21(8):2 684-2 698.
[29]  Lauth M,Toftgrd R.Hedgehog signaling and pancreatic tumor development[J].Advances in Cancer Research,2011,110:1-17.
[30]  Chen H,Zou Y,Yang H,et al.Downregulation of FoxM1inhibits proliferation,invasion and angiogenesis of HeLa cells in vitro and in vivo[J].International Journal of Oncology,2014,45(6):2 355-2 364.
[31]  Shi M,Cui J,Xie K.Signaling of miRNAs-FOXM1in cancer and potential targeted therapy[J].Current Drug Targets,2013,14(10):1 192-1 202.
[32]  Teh MT,Wong ST,Neill GW,et al.FOXM1is a downstream target of Gli1in basal cell carcinomas[J].Cancer Research,2002,62(16):4 773-4 780.
[33]  Guan P,Chen H,Li HJ,et al.Expression and significance of FOXM1in human cervical cancer:a tissue micro-array study[J].Clinical&Investigative Medicine Medecine Clinique Et Experimentale,2011,34(1):1-7.
[34]  Büller NV,Rosekrans SL,Westerlund J,et al.Hedgehog signaling and maintenance of homeostasis in the intestinal epithelium[J].Physiology,2012,27(3):148-155.
[35]  Nusslein-Volhard C,Wieschaus E.Mutations affecting segment number and polarity in Drosophila[J].Nature,1980,287(5 785):795-801.
[36]  Long B,Wang LX,Zheng FM,et al.Targeting GLI1Suppresses Cell Growth and Enhances Chemosensitivity in CD34+Enriched Acute Myeloid Leukemia Progenitor Cells[J].Cellular Physiology&Biochemistry,2016,38(4):1 288-1 302.
[37]  Bella L,Zona S,Moraes GND,et al.FOXM1:A key oncofoetal transcription factor in health and disease[J].Seminars in Cancer Biology,2014,29:32-39.
[38]  Lam EW,Brosens JJ,Gomes AR,et al.Forkhead box proteins:tuning forks for transcriptional harmony[J].Nature Reviews Cancer,2013,13(7):482-495.
[39]  Lamouille S,Xu J,Derynck R.Molecular mechanisms of epithelial-mesenchymal transition[J].Nature Reviews Molecular Cell Biology,2014,15(3):178-196.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133