Immunomodulatory effect of adriamycin in mouse 4T1 model of breast cancer / 生物工程学报

To explore the immunomodulatory effect of adriamycin on 4T1 breast cancer. We used a tandem mass tag-based quantitative proteomic method to detect differential proteins in breast cancer tissues, and multiple bioinformatics databases to analyze the differentially expressed proteins in the proteome. Also, we used enzyme-linked immunosorbent assay to detect the effects of adriamycin on helper T cells 1 and 2 in breast cancer tissues, and flow cytometry to detect CD4+ T cells, CD8+ T cells and regulatory T cells. We discovered the immunomodulatory targets of adriamycin in differential proteins. In total 170 differential proteins were significantly up-regulated, whereas 58 were markedly down-regulated. In addition, 73 proteins were involved in immune regulation. Kyoto encyclopedia of genes and genomes enriched important protein pathways related to cytokines and factor receptors, interleukin 17 pathway and cancer transcriptional regulatory pathways. These pathways and important differential proteins related to immunomodulatory functions were ultimately regulated by adriamycin on CD4+ T cells, CD8+ T cells and regulatory T cells, thereby affecting the prognosis of breast cancer. Moreover, adriamycin significantly increased interleukin 2, CD4+ T and CD8+ T (P<0.01) and markedly reduced regulatory T cells (P<0.05). The function of adriamycin against triple-negative breast cancer was closely related to the immunoregulation process of the differential proteins Ighm, Igkc, S100A8, S100A9 and Tmsb4x. Adriamycin could regulate the content of helper T cells 1 cytokines, CD4+ T and CD8+ T lymphocytes in breast cancer and reduce the number of regulatory T cells to produce immunomodulatory effects.

In vivo and in vitro effects of Miao Medicine Indigofera stachyoides extracts on breast cancer 4T1 cells / 中草药

Objective To study the effects of Indigofera stachyoide extracts on the breast cancer cells (4T1) in vivo and in vitro. Methods MTT method was used to detect the antitumor activity of I. stachyoides extracts in 4T1cells in vitro, the inhibition rate of cell proliferation, and half inhibition concentration (IC50). The established mice model with 4T1 solid tumor were randomly divided into model, extracts of I. stachyoides (petroleum ether phase, ethyl acetate phase, n-butanol phase, water phase, and ethanol extracts) groups, and cisplatin group. After being administered for 15 d, mice body weight and victera index were measured; The observation of tumor pathology and the calculation of tumor inhibition rate were performed. Results IC50 of ethyl acetate phase, n-butanol phase, ethanol extracts of I. stachyoides on 4T1 cells in vitro reached 228.9, 323.4, and 322.6 μg/mL, respectively. The tumor inhibition rates of petroleum ether phase, ethyl acetate phase, n-butanol phase, water phase, and ethanol extract of I. Stachyoides, and cisplatin group on 4T1 mice were (55.88 ± 6.68)%, (66.67 ± 14.32)%, (65.71 ± 12.38)%, (53.81 ± 16.17)%, (43.73 ± 25.73)%, and (76.85 ± 11.38)%, respectively. In the different extraction parts of I. stachyoide, the petroleum ether group had the effects of reducing the spleen index, increasing the thymus index and IL-2 level, and the ethyl acetate part was the best partaccording to tumor volume and the tumor suppressor rate. HE staining showed that the tumor cells in petroleum ether extract group were less than that in the model group, the cell arrangement was loose, the pathological mitosis and tumor cell infiltration were less than those of model group, and there was a small amount of lymphocytes and macrophages infiltration. Conclusion The extracts of I. stachyoides can inhibit the growth of 4T1 tumor cells in vivo and in vitro, and its mechanism may enhance the body immunity, so as to inhibit the tumor growth.

Effect of M-CSF and GM-CSF on migration and VEGF-A expression of breast cancer cell line 4T1 / 肿瘤研究与临床

Objective To investigate the effect of M-CSF and GM-CSF on migration and expression of VEGF-A in breast cancer cell line 4T1.Methods Real-time PCR was used to detect VEGF-A mRNA expression in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF or GM-CSF.Ability of migration and metastasis of 4T1 cells were analyzed by scratch and Transwell assays.Results The relative expression of VEGF-A mRNA at 12 h and 24 h in 4T1 cells treated by 5 ng/ml or 10 ng/ml M-CSF were 17.81±2.49 and 17.48± 5.43,5.15±2.59 and 5.45±4.28,respectively,while those treated by GM-CSF were 9.77±2.39 and 7.61±2.80,6.53±2.41 and 6.30±2.89,respectively.M-CSF and GM-CSF can promote the expression of VEGF-A in 4T1 cells (P ＜ 0.05).The relative expression of VEGF-A was higher in 4T1 cells treated for 12 h than that for 24 h (P ＜ 0.01).M-CSF,GM-CSF and VEGF-A can promote metastasis of 4T1 cells (all P ＜ 0.05),whereas no gross migration of 4T1 cells was showed by VEGF-A treatment.Conclusion M-CSF and GM-CSF can promote the migration and expression of VEGF-A in breast cancer cell line 4T1.