The purity and focus of isolated total RNA was measured by ultra-violet spectrophotometry. Before getting used in the experiment, the cells were washed three times in PBS, added Annexin V/PI stored in 4 C, stood at room temperature without light for 3 min, and were filtered in 300 mesh filter traps. Flow cytometry was used to evaluate cell apoptosis. 1. 8 Reverse transcribed MAPK activation quantitative PCR detection of IGF 1R, PDGFA, NGF, NF T, and JNK2 mRNA expression in primary breast cancer cells and breast cancer cell line MDA MB 231 Cells were inoculated into four 75 mL flasks and cultured for 48 h in RPMI 1640 culture medium plus 10 % fetal bovine serum. After removing the original medium, cells were treated for 48 h with medications as described in 1. 5. Total RNA in all experimental groups was isolated with RNAiso Plus following instructions. The cDNA was then reverse transcribed based on the guidelines within the reagent kit and amplified via PCR with w actin and glyceraldehyde 3 phosphate dehydrogenase Skin infection as interior consults. Primer design computer software Primer 5. 0 from Shanghai Biotechnology was used to create the primer. The primer sequence was the following. The optical band concentration was recorded and analyzed using the Gel Analysis System. Recognition of general protein strength was displayed within the percentage of the optical protein band concentration towards the internal gene t actin. 1. 10 Detection of protein expression in xenografted cyst tissue in nude mice by immunohistochemistry Xenografted cancers from diminished nude mice were obtained for immunohistochemical analysis. The look of brown granules in the cytoplasm was considered positive for protein. The integral optical concentration of slides in each group was examined via Image Pro Plus. All data were analyzed with SPSS 18. 0 and represented as. A totally randomly developed E2 conjugating analysis of variance was used to examine the data among groups, and variations of P 0. 05 were considered statistically significant. 33. 1 Growth, morphology, and appraisal of breast carcinoma cells The cultured breast carcinoma cells showed secure proliferation after two weeks by staying with the wall in long taxi shapes, although some interstitial cells showed in polygon extending growth, often the cell fragments and dross included there. Differential adhesion was used to eliminate the interstitial cells and fibroblasts. Breast carcinoma cells were those whose cell stability reached 3 months as found by trypan blue stain and that achieved positive results for cytoplasmic glycoprotein in immunocytochemical staining. Major breast carcinoma cells were treated with UTI, TXT, or UTI TXT for 72 h, and the results showed that UTI, TXT, and UTI TXT significantly inhibited the proliferation of breast carcinoma cells. These inhibitory effects were statistically significant in contrast to the control group. Additionally, the inhibitory effect was increased after prolonged treatment, which shows a time dependent effect. UTI, TXT, and UTI TXT also dramatically inhibited the growth of MDA MB 231 cells in contrast to the control group, and the inhibitory effect was enhanced after extended treatment.