文档介绍：华中科技大学
硕士学位论文
EGFR、ADAM9在胰腺癌Panc-1微球体细胞中表达的研究
姓名:洪晓泉
申请学位级别:硕士
专业:普通外科
指导教师:王欣
2011-04
华中科技大学硕士学位论文
EGFR、ADAM9 在胰腺癌 Panc-1 微球体细胞中表达的研究

华中科技大学同济医学院附属同济医院胆胰外科
硕士研究生:洪晓泉
指导教师:王欣副教授

中文摘要
目的通过无血清培养法体外培养胰腺癌微球体细胞,验证其肿瘤干细胞的富集效
果,检测 EGFR 和 ADAM9 在胰腺癌微球体细胞中的表达。
方法运用无血清条件培养基,悬浮培养胰腺癌 Panc-1 细胞,并连续传代培养。将
培养好的微球体接种于含血清培养基及胶原底物培养条件下进行分化诱导。收集微球
体细胞及分化细胞,流式细胞术检测侧亚群细胞(side population,SP)比例;Real-time
PCR 及 Western blot 检测 EGFR 和 ADAM9 mRNA 和蛋白的表达。
结果成功培养出胰腺癌 Panc-1 细胞微球体,并能在体外连续传代;分化诱导后重新
贴壁生长。微球体细胞和分化细胞中 SP 比例分别为(±)%和(±)
% (P<)。微球体细胞与分化细胞相比,EGFR 及 ADAM9 mRNA 表达分别上调了
约和倍(P<);蛋白表达分别上调了约倍和倍(P<)。
结论微球体培养能富集胰腺癌干细胞,ADAM9 可能通过 EGFR 信号通路在胰腺癌
的发生发展中起重要作用,有望成为胰腺癌新的治疗靶点。
关键词胰腺癌干细胞;微球体细胞;EGFR;ADAM9

1
华中科技大学硕士学位论文
Expression of EGFR and ADAM9 in Pancreatic Cancer Panc-1
Cell Spheres

Department of Pancreatic-Biliary Surgery, Tongji Hospital, Tongji Medical College,
Huazhong University of Science and Technology
post-graduate: Hong Xiaoquan
Tutor: Pro. Wang Xin

ABSTRACT
Objective The theory of cancer stem cells(CSCs) offer us a new thought about tumors,
more and more kinds of cancer stem cells were isolated and identified form corresponding
tissue. Our aim was to isolate and identify CSCs in Pancreatic cancer cell lines Panc-1 with
serum-free medium(SFM)including epidermal growth factor(EGF), basic fibroblast growth
factor(bFGF), insulin and B27 supplement, and to detect the expression level of EGFR and
ADAM9 by Real-time PCR and Western blot.
Methods Self-renewal assay, differentiation assay, detection of side population (SP)
cells were performed to characterize CSCs in Panc-1 mammospheres culturing with
serum-free medium. Moreover, Real-time PCR and Western blot were used to detect the
expression of EGFR and ADAM9.
Results Panc-1 passaged and formed mammospheres in the absence of serum. Cell
spheres differentiation occurred when serum was su