Analysis of gene expression associated cancer invasion and metastasis using in situ RT-PCR method

1.我们研究了尿激酶型纤溶酶原激活物（uPA）、尿激酶型纤溶酶原激活物受体（uPAR）和纤溶酶原激活物抑制剂-1，2（派-1，派-2）在膀胱癌中的表达，以确定其基因表达在膀胱癌侵袭和进展中的作用。uPA、uPAR、派-1和派-2 mRNA在浸润性肿瘤中的表达水平明显增高，且与组织学分级相关。分析膀胱癌组织中p53基因的表达及其与膀胱癌临床病理特征的关系。浸润性肿瘤表达的p53水平明显高于浅表性肿瘤。以上结果表明，uPA、uPAR、派-1、派-2和PAI-1在膀胱癌的发生发展中起重要作用.本研究采用zelatin酶谱法研究了MMP-2的活化形式的表达，以确定其在尿路上皮癌中的作用。MMP-2的活化形式的表达水平在浸润性肿瘤组织中显著较高。高水平的MMP-2活化形式与缩短的病程特异性生存期密切相关。这些结果提示MMP-2的活化形式在尿路上皮癌的侵袭中起重要作用.建立了非侵袭性和侵袭性尿路上皮癌细胞系，并将侵袭性尿路上皮癌细胞系移植到SCID小鼠膀胱内。我们分析了MT 1-MMP和MMP-2 mRNA在浸润性尿路上皮癌细胞系转移到SCID小鼠膀胱形成的膀胱肿瘤组织中的定位。MT 1-MMPmRNA主要表达于肿瘤的进展期，MMP-2 mRNA主要表达于肿瘤间质细胞。此外，我们现在正在分析从手术中获得的组织中的mRNA定位。

1. we investigated the expression of urokinase-type plasminogen activator (uPA), urokinase-type plasminogen activator receptor (uPAR), and plasminogen activator inhibitor-1, 2 (PAI-1, PAI-2) in bladder cancer to determine the role of their gene expression in invasion and progression of bladder cancer. Expression levels of uPA, uPAR, PAI-1 and PAI-2 mRNA was significantly higher in invasive tumor, and was correlated with histological grade. We also analyzed the gene expression of SPARC in bladder cancer and its relationship with clinical-histopathological manifestation. Invasive tumors expressed significantly higher level of SPARC than superficial tumors. These results showed that uPA, uPAR, PAI-1, PAI-2, and SPARC played an important role in the tumor progression of bladder cancer.2. The present study investigated the expression of the activated form of MMP-2 using zelatin zymography in order to define its role in urothelial cancer. Expression levels of activated forms of MMP-2 were significantly higher in invasive tumor tissue. High levels of activated forms of MMP-2 were strongly associated with shortened course-specific survival. These findings suggest that the activated form of MMP-2 plays a significant role in invasion of urothelial cancer.3. We established non invasive and invasive urothelial cancer cell lines, and transplanted invasive urothelial cancer cell line to the bladder of SCID mice. We analyzed the mRNA localization of MT1-MMP and MMP-2 mRNA in bladder tumor tissues formed from invasive urothelial cancer cell line to the bladder of SCID mice. MT1-MMP mRNA was observed in the advanced part of tumor, and MMP-2 mRNA was observed in interstitial cells . Furthermore we are now analysing mRNAs localization in tissues obtained from surgery.