NCI-H446 SCLC Cell Chorioallantoic Membrane Tumor Forming Experiment And The Relationship With MMP-9

Objective Through the NCI-H446 type SCLC cells inoculated into chick embryo chorioallantoic membrane, observe the change of MMP-9 content in the process of tumor formation in embryos and dynamic observation of tumor tissue and tumor adjacent angiogenesis.Methods ①Change culture medium of NCI-H446 SCLC cells in primary culture of 24h and remove impurities and dead cells, the remaining adherent cells were morphologically normal and good. The cells were cultured and the logarithmic phase cells were collected to reserve.②Establish chorioallantoic membrane SCLC cell tumor model.③Using morphological method for dynamic observe of tumor tissue and tumor adjacent angiogenesis.④Collect various time points chick serum and the MMP-9 in serum was detected at different time points in the chick embryo model by enzyme-linked immunosorbent assay(ELISA).Results ①SCLC cell Subculture successful and grow with the increasing concentration of cells can be tumorigenic.② Successfully established NCI-H446 SCLC cell chick chorioallantoic membrane tumor models.③Compared with the control group, NCI-H446 type SCLC cell promote the formation of MMP-9 function.④MMP-9 levels of the SCLC cell counts were positively correlated.⑤MMP-9 content of a positive correlation with the number of blood vessels around the tumor.Conclusion ①SCLC cells of chick embryo chorioallantoic membrane tumor model is easy to operate, easy to control, easy to realize, is a better animal model for the study of tumor cells.②The model is a short experimental period, growth and tumor angiogenesis for dynamic observation of tumor invasion and metastasis of SCLC cells, the study has important significance.②With the increase of planting density of SCLC cells, Chick embryo chorioallantoic membrane gradually stabilizing tumo.When planting concentration reached 4×106mL-1, third days, could lead to content in serum increased significantly in MMP-9 chicken embryos, seventh days to reach the highest value., that SCLC cells can be generated through the MMP-9 to cell metastasis. Inhibition of MMP-9, is an important direction for future research of chemotherapeutic drugs.③There is a close relationship between the content of MMP-9 and tumor surrounding capillaries density. MMP-9 may be based on the angiogenesis promoting effect to aid in tumor cells metastasis. Further research needs to be specific mechanisms of this effect.