| 【Objective】: Compare the biological characteristics of gastric cancer stemcells(CSC-G)and gastric cancer cell line(SGC7901), the aim is to study the invasionand metastasis role in cancer stem cells, and to investigate the angiogenesis of cancerstem cells.【Methods】: To detect the expression level of stem cell markers, such as OCT4,SOX2, c-Myc and Klf4in CSC-G cells and SGC7901cellsby Real-time PCR,Western-blot and cell immunohistochemistry. Sphere formation assay, plate cloningexperiments, transwell and nude mice experiments were test for the tumorigenic andinvasive ability of CSC-G cells and SGC7901cells. Test the mRNA and proteinexpression levels of CD44and E-cadherin to compare the metastasis of two groups ofcells.The L-OHP and5-FU resistance were tested by MTS experiments. The ability ofangiogenesis of CSC-G cells and SGC7901cells were detected by wound healing,transwell migration and vessel formation assay.【Results】: The Real-time PCR showed that the relative expression levels of OCT4,SOX2, Klf4and CD44mRNA were significantly increased in CSC-G compared withSGC7901group (P <0.05), but the difference of c-Myc mRNA relative expressionhad no statistically significant (P>0.05).The mRNA expression of E-cadherininCSC-G was statistically significant down regulation than SGC7901group(P <0.05).The results of Western-blot and immunohistochemistry consistent with Real-timePCR. Spere and plate cloning experiments suggested that the number of clones ofCSC-G cells were much more than SGC7901cell group, and the difference wasstatistically significant (P <0.05). Transwell experiments and wound healing showedthat the ability of migration and invasion of CSC-G cells was enhanced thanSGC7901cells,(P <0.05). MTS assay showed that the IC50to chemotherapeuticdrugs L-OHP and5-FU of CSC-G cells were higher than SGC7901group, and thedifference was statistically significant (P <0.05). In vivo tumorigenicity assay, wefound that the tumorigenesis ability of CSC-G cells were significantly improvedcompared with SGC7901cells, and the difference was statistically significant (P <0.05). Vessel formation assay showed that the ring numbers formed by the CSC-G were much more than SGC7901group, and the difference was statistically significant (P <0.05).【Conclusion】: According to our research, we found that the invasiveness,tumorigenicity and drug resistance of gastric cancer cells was significantly higherthan normal cancer cells, as well as migration and angiogenesis capabilities. Therefore,gastric cancer stem cells play a key role of gastric cancer recurrence, metastasis, anddrug resistance. And CSC-G had an obvious migration and angiogenesis ability,which account for the development and progression of gastric cancer by angiogenesis. |
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