Investigate the role of Podopalanin in lymphatic metastasis on rat C6 glioma cells

• Main Authors: Pei-yun Lin, 林沛雲
• Other Authors: K. Y.Chong
• Format: Others
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/16610203981018583782

碩士 === 長庚大學 === 醫學生物技術暨檢驗學系 === 99 === Tumor metastasis was considered as the major causes of death for cancer patients. Cancer cells acquire the ability to penetrate the walls of lymphatic and/or blood vessels for escape from the primary site. This process is known as lymphatic or hematogeneous metastasis. To date, scientists made promising progress towards inhibiting haematogenous metastasis of cancer patients, but little is known about lymphatic metastasis. Recent studies demonstrated that lymphatic metastatic routes were controlled by specific gene alternation of tumor cells, In additions, Podopalanin (pdpn) was involved in cancer development, lymphatic metastasis and poor prognosis a wide variety of cancer types. In our laboratory previous study indicated that using tail-vein injection of rat C6 glioma cells into nude mice, the cancer cells colonized at lung were isolated for establishing a C6 subline (C6-lung). Simultaneously, the circulating tumor cells were isolated for establishing another C6 subline (C6-blood). When these C6 sublines were inoculated into nude mice subcutaneously, we found that they have quite different in vivo metastatic potential and activity. Interestingly, we also noticed that the pdpn mRNA and protein was up-regulation in metastatic C6 sublines. Therefore, we hypothesize the C6 glioma cell which overexpress pdpn has potential of lymphatic metastasis. To investigate the relationship between pdpn and tumor lymphatic metastasis, we studied: (1) to characterize parental and metastatic from C6 glioma cells in vitro (2) to investigate the pdpn mediated cell physiological in parental and metastatic from C6 glioma cells. Our results shown that compared with parental cells, the C6-lung cells exhibited significant decreasing in cell motility but had a greater migratory capacity, whereas the C6-blood cells displayed moderate decreasing in cell motility. Furthermore, Stable clones overexpressed pdpn shRNA in C6-LG, were obtained by the blasticidin selection, the real-time RT-PCR and Western blot results shown that endogenous pdpn mRNA and protein expression was significantly inhibited from stably pdpn shRNA expressed clones. We also found that the lymphatic metastasis related marker VEGF-C was significant downregulated in pdpn shRNA of C6-blood, but significant upregulated in pdpn shRNA of C6-lung sublines. In conclusion, we demonstrated that pdpn overexpression in C6 subline was associated with tumour lymphatic metastasis, two C6 sublines displayed a totally different cellular physiological phenomena.