细胞侵袭迁移检测（Cell invasion and migration detection）.docVIP

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细胞侵袭迁移检测（Cell invasion and migration detection）.doc

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细胞侵袭迁移检测（Cell invasion and migration detection）

细胞侵袭迁移检测（Cell invasion and migration detection） Colony formation of 1.2.3 soft agar: We observed the effect of PRL-3 and siRNA on the anchorage independent proliferation of colorectal cancer cells by soft agar colony culture method according to the reference method [10] 1.2.4 invasion: collect 48 h transfected cells, the cancer cells to detect Boyden cell model of [11] invasion. Boyden cell membrane of Matrigel cells through the upper chamber to the lower chamber of the membrane surface, the number reflects the invasion ability of cell size. 400 times the number of cells was counted under light microscope through the polycarbonate membrane, to the relative number of invasive cells expressed tumor cell invasion. The number of cells were counted in 5 in the field, taking the average value for statistical processing, each counting 3 samples. The invasion of colorectal cancer cell line 1.2.5 in nude mice: cells were collected without serum DMEM after 2 times washing, accurate count and adjust the cell concentration of 3 * 109/L, the cells were inoculated into nude mice, 0.2 mL/ rats. After 28 d, breaking the neck anatomy of nude mice, nude mice were killed, with 40 g/L formaldehyde fixed collection the organization, production, HE staining, observe the tumor cell metastasis and invasion in vivo. In the.2 soft agar colony forming assay, large intestine cancer HCT116 cells can spontaneously form colonies in vitro in semisolid culture system, whereas the cell growth by PRL-3 and siRNA transfection decreased in a dose-dependent manner (P0.05, Fig. 2) Fig. 2 Effect of PRL-3 siRNA transfection on anchorage independent proliferation of colorectal cancer cells 2.3 effects of PRL-3 siRNA on invasion of colon cancer cells, this study adopts Boyden model method using PRL-3 siRNA transfection on the invasion of colorectal cancer cells showed that: compared with the control group, siRNA group through the cancer cell membrane decreased significantly, and the relationship with concent