Jagged-1在宫颈癌和卵巢癌中表达的初步研究
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摘要
目的：检测Jagged-1蛋白在宫颈鳞状上皮细胞癌(Squamous cell carcinomasSCC)、宫颈腺癌和卵巢浆液性囊腺癌组织中的表达,探讨Jagged-1蛋白在宫颈癌和卵巢癌发生发展中的作用机制。
方法：采用免疫组织化学EnVision法,对30例宫颈SCC、15例宫颈腺癌、14例宫颈正常组织及44例卵巢浆液性囊腺癌、14例正常卵巢组织中Jagged-1蛋白进行检测,用妊娠早期绒毛Jagged-1阳性表达作为阳性对照,用PBS代替一抗作为空白对照,根据肿瘤细胞染色程度以及阳性细胞所占百分比进行综合评分。采用SPSS13.0软件进行统计分析宫颈正常组织、宫颈鳞癌和腺癌中Jagged-1蛋白的表达,以及卵巢正常组织、卵巢浆液性囊腺癌组织中Jagged-1蛋白的表达水平,与病理组织学分级和临床分期等的关系。
结果：
1. Jagged-1蛋白在宫颈癌中的表达：Jagged-1蛋白在宫颈癌中主要表达在癌细胞的细胞核,在宫颈正常组织中阳性着色定位在基底细胞,其阳性表达为黄色或棕黄色颗粒,其在宫颈鳞癌、腺癌、正常组织中阳性表达率分别为66.7%、80.0%、21.4%。鳞癌、腺癌中Jagged-1蛋白的表达均高于正常组织,差异有统计学意义(P<0.05),鳞癌和腺癌相比差异无统计学意义(P>0.05)。
2. Jagged-1蛋白在卵巢癌中的表达：Jagged-1蛋白在卵巢浆液性囊腺癌中表达在癌细胞的细胞浆和细胞核,在卵巢正常组织中阳性定位在部分颗粒细胞。其阳性表达亦为黄色或棕黄色颗粒,在卵巢浆液性囊腺癌中及正常组织中的阳性表达率分别为68.2%和28.6%,差异有统计学意义(P<0.05),在卵巢癌中Jagged-1蛋白的表达与卵巢癌患者年龄、肿瘤大小、临床分期无相关性(P>0.05),但随着卵巢癌病理分级增高,Jagged-1蛋白的阳性表达率增高,在病理2、3级中Jagged-1蛋白阳性表达率79.3%,明显高于1级阳性率(46.4%),G2、G3级与Gl级相比,P<0.05,差异有统计学意义。
结论：
1. Jagged-1蛋白在宫颈SCC和宫颈腺癌组织中均有较高的阳性表达率。
2. Jagged-1蛋白在卵巢浆液性囊腺癌组织中高表达,且病理组织分级越高,Jagged-1阳性表达率越高,提示随着病理分级的增高,其表达上调。
Objective
To study on the expression of Jagged-1 protein in cervical cancer and ovarian serous cystadenocarcinoma, so as to explore Jagged-1 relationship in the mechanism about cervical cancer and ovarian serous cystadenocarcinoma.
Method
En Vision immunohistochemical (IHC) technique was used to assay the expression of Jagged-1 protein in 30 cases of cervical squamous cell carcinomas,15 cases of cervical adenocarcinoma,14 cases of normal cervical tissue and 44 cases of ovarian serous cystadenocarcinoma,14 cases of normal ovarian tissue. The sample of villi in early trimester of pregnancy was used as positive control and PBS replace the first antibody as blank control. According to the staining degree and the percent of positive cell in tumor cell, we divide into glade to be evaluated. The recorded data were analyzed using SPSS software 13.0 version.
Results
1. The expression of Jagged-1 protein in cervical carcinoma:
Jagged-1 protein positive in cervical carcinoma express in cell nucleus of tumor cells. But Jagged-1 protein was less expressed in basal cell in normal tissue of cervix. The positive rate of cervica squamous cell carcinomas, adenocarcinoma, and normal tissue are 66.7%,80.0% and 21.4%, respectively. The expression of Jagged-1 protein in cervical carcinoma was higher apparently than normal tissue. There were significant differences in them. (P<0.05)
2. The expression of Jagged-1 protein in ovarian cancer:
Jagged-1 protein was expressed in cytoplasm and nucleus, and was positive in granular cell in normal tissue of ovary. The expression rate of Jagged-1 protein in the serous cystadenocarcinoma, normal ovarian tissue were 68.2%,28.6%, respectively. There were significant differences in ovarian serous cystadenocarcinoma and normal ovarian tissue (P<0.05). In ovarian cancer group, there was no significant difference for Jagged-1 expression in different age patients groups. And there was no difference among different tumor size and clinical stages. But the percent of Jagged-1 protein enhanced according to pathological grade change. There were significant differences in pathological grade 2 and 3 (79.3%) to grade 1 (46.4%), P<0.05.
Conclusions
1. Jagged-1 protein is positive expressed in both cervical squamous cell carcinomas and adenocarcinoma.
2. Jagged-1 protein is positive expressed highly cystadenoma in ovary. And the expression of Jagged-1 protein is more positive with the pathological grade change. It suggested Jagged-1 is related in the cervical carcinoma and ovarian serous cystadenocarcinoma.
方法：采用免疫组织化学EnVision法,对30例宫颈SCC、15例宫颈腺癌、14例宫颈正常组织及44例卵巢浆液性囊腺癌、14例正常卵巢组织中Jagged-1蛋白进行检测,用妊娠早期绒毛Jagged-1阳性表达作为阳性对照,用PBS代替一抗作为空白对照,根据肿瘤细胞染色程度以及阳性细胞所占百分比进行综合评分。采用SPSS13.0软件进行统计分析宫颈正常组织、宫颈鳞癌和腺癌中Jagged-1蛋白的表达,以及卵巢正常组织、卵巢浆液性囊腺癌组织中Jagged-1蛋白的表达水平,与病理组织学分级和临床分期等的关系。
结果：
1. Jagged-1蛋白在宫颈癌中的表达：Jagged-1蛋白在宫颈癌中主要表达在癌细胞的细胞核,在宫颈正常组织中阳性着色定位在基底细胞,其阳性表达为黄色或棕黄色颗粒,其在宫颈鳞癌、腺癌、正常组织中阳性表达率分别为66.7%、80.0%、21.4%。鳞癌、腺癌中Jagged-1蛋白的表达均高于正常组织,差异有统计学意义(P<0.05),鳞癌和腺癌相比差异无统计学意义(P>0.05)。
2. Jagged-1蛋白在卵巢癌中的表达：Jagged-1蛋白在卵巢浆液性囊腺癌中表达在癌细胞的细胞浆和细胞核,在卵巢正常组织中阳性定位在部分颗粒细胞。其阳性表达亦为黄色或棕黄色颗粒,在卵巢浆液性囊腺癌中及正常组织中的阳性表达率分别为68.2%和28.6%,差异有统计学意义(P<0.05),在卵巢癌中Jagged-1蛋白的表达与卵巢癌患者年龄、肿瘤大小、临床分期无相关性(P>0.05),但随着卵巢癌病理分级增高,Jagged-1蛋白的阳性表达率增高,在病理2、3级中Jagged-1蛋白阳性表达率79.3%,明显高于1级阳性率(46.4%),G2、G3级与Gl级相比,P<0.05,差异有统计学意义。
结论：
1. Jagged-1蛋白在宫颈SCC和宫颈腺癌组织中均有较高的阳性表达率。
2. Jagged-1蛋白在卵巢浆液性囊腺癌组织中高表达,且病理组织分级越高,Jagged-1阳性表达率越高,提示随着病理分级的增高,其表达上调。
Objective
To study on the expression of Jagged-1 protein in cervical cancer and ovarian serous cystadenocarcinoma, so as to explore Jagged-1 relationship in the mechanism about cervical cancer and ovarian serous cystadenocarcinoma.
Method
En Vision immunohistochemical (IHC) technique was used to assay the expression of Jagged-1 protein in 30 cases of cervical squamous cell carcinomas,15 cases of cervical adenocarcinoma,14 cases of normal cervical tissue and 44 cases of ovarian serous cystadenocarcinoma,14 cases of normal ovarian tissue. The sample of villi in early trimester of pregnancy was used as positive control and PBS replace the first antibody as blank control. According to the staining degree and the percent of positive cell in tumor cell, we divide into glade to be evaluated. The recorded data were analyzed using SPSS software 13.0 version.
Results
1. The expression of Jagged-1 protein in cervical carcinoma:
Jagged-1 protein positive in cervical carcinoma express in cell nucleus of tumor cells. But Jagged-1 protein was less expressed in basal cell in normal tissue of cervix. The positive rate of cervica squamous cell carcinomas, adenocarcinoma, and normal tissue are 66.7%,80.0% and 21.4%, respectively. The expression of Jagged-1 protein in cervical carcinoma was higher apparently than normal tissue. There were significant differences in them. (P<0.05)
2. The expression of Jagged-1 protein in ovarian cancer:
Jagged-1 protein was expressed in cytoplasm and nucleus, and was positive in granular cell in normal tissue of ovary. The expression rate of Jagged-1 protein in the serous cystadenocarcinoma, normal ovarian tissue were 68.2%,28.6%, respectively. There were significant differences in ovarian serous cystadenocarcinoma and normal ovarian tissue (P<0.05). In ovarian cancer group, there was no significant difference for Jagged-1 expression in different age patients groups. And there was no difference among different tumor size and clinical stages. But the percent of Jagged-1 protein enhanced according to pathological grade change. There were significant differences in pathological grade 2 and 3 (79.3%) to grade 1 (46.4%), P<0.05.
Conclusions
1. Jagged-1 protein is positive expressed in both cervical squamous cell carcinomas and adenocarcinoma.
2. Jagged-1 protein is positive expressed highly cystadenoma in ovary. And the expression of Jagged-1 protein is more positive with the pathological grade change. It suggested Jagged-1 is related in the cervical carcinoma and ovarian serous cystadenocarcinoma.
引文
[1]Parkin DM.Global cancer statistics in the year 2000[J].Lancet Oncol,2001,2(9): 533-543.
[2]林伟华,张晶.宫颈癌的预防及普查[J].中国实用医药,2007,32：116.
[3]章文华.宫颈癌筛查方法与我国宫颈癌筛查面临的新问题.中华肿瘤杂志[J],2008,30(12)：881—884.
[4]郑怀美主编.现代妇产科.第一版.上海：上海医科大学出版社,2001,444-500
[5]Choi JH,Park JT, Davidson B, et al. Jagged-1 and Notch3 Juxtacrine Loop Regulates Ovarian Tumor Growth and Adhesion. Cancer Res 2008,68 (14): 5716-5723.
[6]Kojika S,Griffin JD.Notch receptors and hetopoieis.Exp Hemtol.2001,29: 1041-1052.
[7]Holland LZ, Rached LA, Tammc R, et al. Chanacyerization and developmental expression of the amphioxus homolog of Notch:Evolution conservation of multiple expreeion domains in amphioxus and vertebrates. Dev Biol,2001,232: 493-507.
[8]齐润姿,曹雪涛.Notch的结构与信号传导.中国肿瘤生物治疗杂志,2002,9(3)：212-214.
[9]Wakamatsu Y, Maynard TM, Weston JA, Fate determination of neural crest cells by NOTCH-mediated lated inhibition and asymmetrical cell division gangliogenesis. Development,2000,127:2811-2821.
[10]Grace E,Robert S, Efthimios Mitsiadis,et al. Human Ligands of the Notch Receptor. American Journal of Pathology,1999,154:789-793.
[11]方志远,邢飞跃.Jagged 1促进肿瘤发生.生物学杂志,2008,25(6)：1-5
[12]Reedijk M,Odoreic S,Chang L,et al.High-level coexpression of JAG 1 and NOTCH 1 is observed in human breast cancer and is associated with poor overall survival[J].Cancer Res,2005,65(18):8530-8537.
[13]姜昕,周建华,邓征浩等.Notchl, Jaggedl和VEGF蛋白在非小细胞肺癌的表达及其意义.中南大学学报(医学版),2007,32(6)：1031-1036.
[14]张秀萍,艾星,姜杰等.膀胱移行细胞癌组织Notch-1和Jagged-1蛋白表达临床意义的探讨.中华肿瘤防治杂志,2009,16(16)1242-1245.
[15]Callahan R,Smith GH.MMTV-induced mammary tumorigenesis:gene discovery,progression to malignanc and cellular pathways. Oncogene, 2000,19(8):992-1001.
[16]Gerard F. Hoyne. Notch signaling in the immune system. J. Leukoc. Biol.2003, 74:201-208.
[17]De Falco M, Cobellis L, Giraldi D, et al. Expression and distribution of notch protein members in human placenta throughout pregnancy. Placenta.2007, 28(2-3):118-206.
[18]Demarest RM, Ratti F, Capobianco AJ. It's T-ALL about Notch. Oncogene, 2008,27:5082-5091.
[19]鲁照明,刘红涛,许培荣等.Notchl基因在食管鳞癌细胞株EC9706中的表达及其对细胞凋亡的影响.癌症Chinese Journal of Cancer,2007,26(10): 1074-1079.
[20]Djonov V,Andres A C,Ziemiecki A. Vascular remodelling during the normal and malignant life cycle of the mammary gland[J]. Microsc Res Tech,2001,52 (2):182-189.
[21]Gray GE,Mann RS,Mitsiadis E,et al.Human Ligands of the Notch Receptor. American Journal of Pathology,1999,154 (3) 785-794.
[22]周恩昆,陈默,方廖琼.Notch信号通路与乳腺肿瘤发生关系的研究进展.中华肿瘤防治杂志,2008,15(10)：787-790.
[23]Leong K G, Karsan A. Recent insights into the role of Notch signaling in tumorigenesis [J].Blood,2006,107(6):2223-2233.
[24]Krantz I D,Colliton R P,Genin A,et al. Spectrum and Frequency of Jaggedl (JAG1) Mutations in Alagille Syndrome Patients and Their Families.Am.J.Hum.Genet,1998,62:1361-1369.
[25]Tohda S,Nara N.Expression of Notchl and Jaggedl proteins in acute myeloid leukemia cells[J].Leuk Lymphoma,2001,42(3):467-472.
[26]Reedijk k M,Odorcic S,Chang L,et al.High-level coexpression of JAG1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival[J]. Cancer Res,2005,65(18):8530-8537.
[27]Reedijk M,Pinnaduwage D,Dickson BC,et al.JAGl expression is associated with a basal phenotype and recurrence in lvmph nodenegative breast cancer[J]. Breast Cancer Res Treat,2008,111(3):439-448.
[28]Thurston G,Noguera-Troise I, Yancopoulos GD. The Delta paradox:DLL4 blockade leads to more tumour vessels but less tumour growth. Nat Rev Cancer,2007,7(5):327-331.
[29]董学斌,纪春岩,马道新,等.Notch信号在人类乳腺癌中的作用.中华肿瘤杂志,2007,29(6)：425—428.
[30]Rodilla V,Villanueva A,Obrador-Hevia A,et al.Jaggedl is the pathological link between Wnt and Notch pathways in colorectal cancer.PNAS,2009,106 (15):6316-6320.
[31]Reedijk M,Odorcic S,Zhang H,et al.Activation of Notch signaling in human colon adenocarcinoma. Int J Oncol.2008,33(6):1223-1229.
[32]Oda T,Elkahloun AG,Pike BL,et al.Mutations in the human JAG1 gene are responsible for Alagille syndrome.Nat Genet,1997,16:235-242.
[33]Regan G N,Majesky M W.Building a Vessel Wall with Notch Signaling. Circ Res.2009,104(4):419-421.
[34]徐洪雨,李宝杰,王瑞峰,等.Notch/Jagged信号在肝部分切除后肝再生中的表达.胃肠病学和肝病学杂志,2007,16(5)：465-467.
[35]Zeng Q,Li S,Chepeha DB,et al.Crosstalk belween tumor and endothelial cells promotes tumor angiogenesis by MAPK activation of Notch signaling[J]. Cancer Cell,2005,8(1):13-23.
[36]赵慧等,Jaggedl与肿瘤的研究进展.医学综述,2009,15(12)：1799-1801.
[37]Reedijk M,Odorcic S,Chang L,et al.High-level coexpression of JAG 1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival.Cancer Res,2005,65(18):8530-8537.
[38]廖宇圣,吴杰,王萍,等.Notch-1、Jagged-1及p-mTOR在胰腺癌中的表达及意义.胃肠病学和肝病学杂志,2009,18(8)688-691.
[39]Gao J, Chen C, Hong L, et al. Expression of Jaggedl and its association with hepatitis B virus X p rotein in hepatocellular carcinoma[J]. Biochem Biophys Res Commun,2007,356 (2):341-347.
[40]Santagata S, Demichelis F, Riva A, et al. JAGGED1 Expression Is Associated with Prostate Cancer Metastasis and Recurrence. Cancer Res 2004,64 (19): 6854-6857.
[41]刘玉和,于春艳,马志明,等.Notch表达在Vk3诱导的人宫颈癌Hela细胞损伤中的作用及其机制的研究.中国妇幼保健杂志,2008,23(36)5207-5209.
[42]Karthikeyan V,Mark P,Rekha VK,et al.Papillomavirus-Mediated Neoplastic Progression Is Associated with Reciprocal Changes in Jaggedl and Manic Fringe Expression Linked to Notch Activation. journal of virology,2004,78 (16):8687-8700.
[2]林伟华,张晶.宫颈癌的预防及普查[J].中国实用医药,2007,32：116.
[3]章文华.宫颈癌筛查方法与我国宫颈癌筛查面临的新问题.中华肿瘤杂志[J],2008,30(12)：881—884.
[4]郑怀美主编.现代妇产科.第一版.上海：上海医科大学出版社,2001,444-500
[5]Choi JH,Park JT, Davidson B, et al. Jagged-1 and Notch3 Juxtacrine Loop Regulates Ovarian Tumor Growth and Adhesion. Cancer Res 2008,68 (14): 5716-5723.
[6]Kojika S,Griffin JD.Notch receptors and hetopoieis.Exp Hemtol.2001,29: 1041-1052.
[7]Holland LZ, Rached LA, Tammc R, et al. Chanacyerization and developmental expression of the amphioxus homolog of Notch:Evolution conservation of multiple expreeion domains in amphioxus and vertebrates. Dev Biol,2001,232: 493-507.
[8]齐润姿,曹雪涛.Notch的结构与信号传导.中国肿瘤生物治疗杂志,2002,9(3)：212-214.
[9]Wakamatsu Y, Maynard TM, Weston JA, Fate determination of neural crest cells by NOTCH-mediated lated inhibition and asymmetrical cell division gangliogenesis. Development,2000,127:2811-2821.
[10]Grace E,Robert S, Efthimios Mitsiadis,et al. Human Ligands of the Notch Receptor. American Journal of Pathology,1999,154:789-793.
[11]方志远,邢飞跃.Jagged 1促进肿瘤发生.生物学杂志,2008,25(6)：1-5
[12]Reedijk M,Odoreic S,Chang L,et al.High-level coexpression of JAG 1 and NOTCH 1 is observed in human breast cancer and is associated with poor overall survival[J].Cancer Res,2005,65(18):8530-8537.
[13]姜昕,周建华,邓征浩等.Notchl, Jaggedl和VEGF蛋白在非小细胞肺癌的表达及其意义.中南大学学报(医学版),2007,32(6)：1031-1036.
[14]张秀萍,艾星,姜杰等.膀胱移行细胞癌组织Notch-1和Jagged-1蛋白表达临床意义的探讨.中华肿瘤防治杂志,2009,16(16)1242-1245.
[15]Callahan R,Smith GH.MMTV-induced mammary tumorigenesis:gene discovery,progression to malignanc and cellular pathways. Oncogene, 2000,19(8):992-1001.
[16]Gerard F. Hoyne. Notch signaling in the immune system. J. Leukoc. Biol.2003, 74:201-208.
[17]De Falco M, Cobellis L, Giraldi D, et al. Expression and distribution of notch protein members in human placenta throughout pregnancy. Placenta.2007, 28(2-3):118-206.
[18]Demarest RM, Ratti F, Capobianco AJ. It's T-ALL about Notch. Oncogene, 2008,27:5082-5091.
[19]鲁照明,刘红涛,许培荣等.Notchl基因在食管鳞癌细胞株EC9706中的表达及其对细胞凋亡的影响.癌症Chinese Journal of Cancer,2007,26(10): 1074-1079.
[20]Djonov V,Andres A C,Ziemiecki A. Vascular remodelling during the normal and malignant life cycle of the mammary gland[J]. Microsc Res Tech,2001,52 (2):182-189.
[21]Gray GE,Mann RS,Mitsiadis E,et al.Human Ligands of the Notch Receptor. American Journal of Pathology,1999,154 (3) 785-794.
[22]周恩昆,陈默,方廖琼.Notch信号通路与乳腺肿瘤发生关系的研究进展.中华肿瘤防治杂志,2008,15(10)：787-790.
[23]Leong K G, Karsan A. Recent insights into the role of Notch signaling in tumorigenesis [J].Blood,2006,107(6):2223-2233.
[24]Krantz I D,Colliton R P,Genin A,et al. Spectrum and Frequency of Jaggedl (JAG1) Mutations in Alagille Syndrome Patients and Their Families.Am.J.Hum.Genet,1998,62:1361-1369.
[25]Tohda S,Nara N.Expression of Notchl and Jaggedl proteins in acute myeloid leukemia cells[J].Leuk Lymphoma,2001,42(3):467-472.
[26]Reedijk k M,Odorcic S,Chang L,et al.High-level coexpression of JAG1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival[J]. Cancer Res,2005,65(18):8530-8537.
[27]Reedijk M,Pinnaduwage D,Dickson BC,et al.JAGl expression is associated with a basal phenotype and recurrence in lvmph nodenegative breast cancer[J]. Breast Cancer Res Treat,2008,111(3):439-448.
[28]Thurston G,Noguera-Troise I, Yancopoulos GD. The Delta paradox:DLL4 blockade leads to more tumour vessels but less tumour growth. Nat Rev Cancer,2007,7(5):327-331.
[29]董学斌,纪春岩,马道新,等.Notch信号在人类乳腺癌中的作用.中华肿瘤杂志,2007,29(6)：425—428.
[30]Rodilla V,Villanueva A,Obrador-Hevia A,et al.Jaggedl is the pathological link between Wnt and Notch pathways in colorectal cancer.PNAS,2009,106 (15):6316-6320.
[31]Reedijk M,Odorcic S,Zhang H,et al.Activation of Notch signaling in human colon adenocarcinoma. Int J Oncol.2008,33(6):1223-1229.
[32]Oda T,Elkahloun AG,Pike BL,et al.Mutations in the human JAG1 gene are responsible for Alagille syndrome.Nat Genet,1997,16:235-242.
[33]Regan G N,Majesky M W.Building a Vessel Wall with Notch Signaling. Circ Res.2009,104(4):419-421.
[34]徐洪雨,李宝杰,王瑞峰,等.Notch/Jagged信号在肝部分切除后肝再生中的表达.胃肠病学和肝病学杂志,2007,16(5)：465-467.
[35]Zeng Q,Li S,Chepeha DB,et al.Crosstalk belween tumor and endothelial cells promotes tumor angiogenesis by MAPK activation of Notch signaling[J]. Cancer Cell,2005,8(1):13-23.
[36]赵慧等,Jaggedl与肿瘤的研究进展.医学综述,2009,15(12)：1799-1801.
[37]Reedijk M,Odorcic S,Chang L,et al.High-level coexpression of JAG 1 and NOTCH1 is observed in human breast cancer and is associated with poor overall survival.Cancer Res,2005,65(18):8530-8537.
[38]廖宇圣,吴杰,王萍,等.Notch-1、Jagged-1及p-mTOR在胰腺癌中的表达及意义.胃肠病学和肝病学杂志,2009,18(8)688-691.
[39]Gao J, Chen C, Hong L, et al. Expression of Jaggedl and its association with hepatitis B virus X p rotein in hepatocellular carcinoma[J]. Biochem Biophys Res Commun,2007,356 (2):341-347.
[40]Santagata S, Demichelis F, Riva A, et al. JAGGED1 Expression Is Associated with Prostate Cancer Metastasis and Recurrence. Cancer Res 2004,64 (19): 6854-6857.
[41]刘玉和,于春艳,马志明,等.Notch表达在Vk3诱导的人宫颈癌Hela细胞损伤中的作用及其机制的研究.中国妇幼保健杂志,2008,23(36)5207-5209.
[42]Karthikeyan V,Mark P,Rekha VK,et al.Papillomavirus-Mediated Neoplastic Progression Is Associated with Reciprocal Changes in Jaggedl and Manic Fringe Expression Linked to Notch Activation. journal of virology,2004,78 (16):8687-8700.