Src的過度表現與尿路上皮癌細胞的上皮-間質轉化和侵襲偽足形成及對化療藥物抗藥性及幹細胞特性有關
葉碧雯1,2、尤亮恩1,2、李經家1,2、李威明1,2,4、李香瑩1,2,5、柯宏龍1,2,3、李健逢6、吳文正1,2,3*
高雄醫學大學 泌尿科1，高雄醫學大學附設醫院 泌尿科2，高雄醫學大學 醫學研究所3，衛福部屏東醫院 泌尿科4，高雄市立大同醫院 泌尿科5，台南奇美醫學中心 病理科6
Overexpression of SRC is associated with epithelial mesenchymal transition, invadopodia formation, drug resistant and stemness characters in urothelial carcinoma
Bi-Wen Yeh1,2, Liang-En Yu1,2, Ching-Chia Li1,2, Wei-Ming Li1,2,4, Hsiang-Ying Lee1,2,5, Hung-Lung Ke1,2,3, Chien-Feng Li6 and Wen-Jeng Wu1,2,3*
Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung1; Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung2; Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung3; Department of Urology, Ministry of Health and Welfare Pingtung Hospital, Pingtung4; Department of Urology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung5; Department of Pathology, Chi Mei Medical Center, Tainan, Taiwan6
Purposes: Development of cancer cells invasiveness, drug resistance and stem-cell characters are the major problem lead to cancer treatment failure. However, the encompassed molecular mechanisms underlying these processes are complexity and not fully understood. The proto-oncogene c-Src (Src) is a nonreceptor tyrosine kinase whose expression and activity is correlated with advanced malignancy and poor prognosis in a variety of human cancers. Src interacts with multiple RTKs and further triggers multiple cell signaling pathways to promote proliferation, survival, adhesion, migration/invasion and metastatic spread. However, the role Src involved in the progression and metastasis in UC is not yet concluded. Our study was designed to specify the roles of Src in the maintenance of stemness properties in UC cells and to elucidate the correlation of Src expression and prognosis in patients with UC.
Patients and Methods: Src expression in UC cell lines was analyzed by RT-PCR and western blotting. In vitro characterizations of the cellular function of recombinant SRC in epithelial-mesenchymal transition (EMT) and tumorigenic behaviors were evaluated by trans-well assay and colony formation assay, respectively. The role of Src signaling on regulates invadopodia formation and ECM degradation was assessed by gelatin matrix degradation assay. The role of Src signaling on regulates stemness and drug resistance markers were assessed by side population flow cytometry assay. Forty-eight UTUC NT paired specimens were analyzed for Src expression by real-time PCR.
Results and Conclusion: Overexpression of Src increased migration/invasion and ECM degradation capacity (~7 folds) in UC cells. Besides, up-regulation of Src can increase side population characters and increased stemness markers (sox2 and ABCG2) positive cell presented in UC cell lines. Enforced expression of Src is sufficient to up-regulated of EMT markers (vimentin, twist, slug) expression, cancer stemness (cd133 and sox2) and drug resistance related proteins (ABCG2 and MRP1) expression that resulting in increasing invasiveness of UC cells. Collectively, it demonstrated that Src overexpression is associated with an aggressive biological behavior of UC cells. Src expression status represents a prognostic prediction for patients with UTUC after surgery. Further investigating the potential therapeutic role of Src inhibitors in the treatment of UC/UTUC is ongoing.