Mouse Model for Tumor Transplantation (TT)

1.Collect the logarithmic growth phase of esophageal cancer cell line( TE-1), wash with PBS, a total of 5 * l07 cell suspension per milliliter, inoculated subcutaneously in nude mice subcutaneous (0.2 ml/per mice) and then feed in the SPF environment.
2. Regular observation of tumor growth in nude mice, and use vernier caliper to measure the tumor size and draw the tumor growth curve.
3. About 21 days, mice were sacrificed and the tumor tissue removed in l0% formalin fixed, paraffin embedded sections, conventional HE staining and immunohistochemical staining.

1. Cancer tumor growth:
To observe the changes in tumor growth and build tumor growth curve: the size of tumor was measured 2 times per week, the tumor volume was calculated according to the formula, V=Π/6[(A+B)／2]^3 (A, B respectively for tumors of the long diameter and short diameter.

2. HE staining:
The tumor cells become false palisading, oval, irregular shape, cytoplasm stained red or violet red nucleus in violet or purple red, irregularly shaped, oval, unequal in size, clear membrane, chromatin with unequal thickness and deep staining. The tumor cells were arranged densely, unequal in size, uniform distribution of patchy; tumor tissue showed visible focal necrosis, visible cell debris and debris, a small amount of inflammatory cell infiltration, tumor and normal tissue but no clear demarcation, capsule formation.
3. VEGF and CD31 immunohistochemical staining:
VEGF-C and CD31 promote angiogenesis in angiogenesis. Immunohistochemistry results showed that VEGF-C and CD31 positive cells and positive vessels were expressed in tumor tissues

4. Fluorescent quantitative PCR (Q-PCR) detection:
Take control of oesophagus and tumorigenic mouse tumor to make the Q-PCR detection, detection index: VEGF-C, Lymphatic vessel endothelial hyaluronan receptor 1(LYVE-1), GAPDH as reference.

SPSS software is used for statistical analysis, measurement data to mean ± standard deviation (x ±ｓ), using t test and single factor analysis of variance for group comparison , P<0.05 indicates there was a significant difference, P<0.01 indicates there are very significant differences.