ABSTRACT
BACKGROUND: Ectodermal mesenchymal stem cells can differentiate into other types of cells in a specific external environment. The expression of some specific proteins during differentiation may directly affect the direction of their fate. OBJECTIVE: To investigate the expression of tissue transgiutaminase 2 in rat nasal mucosal mesenchymal stem cells during osteoblast differentiation and neuron-like differentiation. METHODS: Rat nasal mucosal mesenchymal stem cells were isolated and cultured by tissue adherence method, and differentiated into neuroblast-like cells and osteoblasts. Immunofluorescence was used to detect the expression of neuro-related proteins GAP-43, TUBB3 and bone-associated proteins OCN and COLI at different time points. Western blot assay was used to detect neuro-related proteins GAP-43, TUBB3, bone-associated protein OCN and COLI, neurotrophic factor NT-3, BDNF, extracellular matrix FN, LN, and tissue transgiutaminase 2. RESULTS AND CONCLUSION: (1) After 3 days of neurogenic induction, immunofluorescence showed that GAP-43 and TUBB3 expressed positively. At 7 days after osteogenic induction, COLI and OCN were positive under immunofluorescence staining. (2) At 3, 5, 7 days after neurogenic induction, the expression levels of nerve cell-associated proteins GAP-43, TUBB3 and tissue transgiutaminase 2 were increased with time detected by western blot assay. At 7 and 14 days after osteogenic induction, the levels of osteoblast-associated proteins COLI, OCN and tissue transgiutaminase 2 were increased with time. (3) At 7 days after neurogenic induction, western blot assay showed that the levels of NT-3, BDNF and tissue transgiutaminase 2 were higher than those in the non-induction group. At 14 days after osteogenic induction, the expression levels of FN, LN and tissue transgiutaminase 2 in the extracellular matrix were higher than those in the non-induction group. (4) To conclude, tissue transgiutaminase 2 may regulate the differentiation of nasal mucosal mesenchymal stem cells into neuron-like cells and osteoblasts. Key words: nasal mucosal mesenchymal stem cells; osteoblasts; neural-like cells; tissue transgiutaminase 2 Funding: the National Natural Science Foundation of China, No. 81571830 (to ZZJ); the Clinical Medicine Science and Technology Development Project of Jiangsu University, No. JLY20180040 (to ZZJ).
ABSTRACT
OBJECTIVE: To explore the mechanisms of Runing Recipe II (a recipe composed of traditional Chinese herbs) in inhibiting the growth of breast cancer by observing its effects on the expressions of p53 and ras oncogene proteins and cell cycle of the transplanted Ca761 breast cancer in mice. METHODS: We established the breast cancer model by transplanting Ca761 cells in mice. The mice were randomly divided into 4 groups: normal saline control group, CTX-treated group, Runing Recipe II-treated group, and Runing Recipe II and CTX-treated group, with 12 mice in each group. We detected the cell cycle of the cancer cells in the mice's transplanted tumor with flow cytometry and measured the expressions of p53 and ras oncogene proteins in the transplanted tumor with immunohistochemical methods. RESULTS: The percentages of tumor cells in S-phase of the Runing Recipe II treated group, CTX-treated group and Runing Recipe II and CTX-treated group were significantly lower than that of the normal saline control group respectively (P<0.05). The percentage of tumor cells in G(0)-G(1) phase of the Runing Recipe II treated group was lower than that of the CTX-treated group (P<0.05), while the percentage of tumor cells in G(2)-M phase was higher than that of the CTX-treated group. The immunoreactive scores (IRSs) of p53 in the Runing Recipe II treated group and Runing Recipe II and CTX-treated group were significantly lower than that in the normal saline control group respectively (P<0.05). The effect of CTX on the expression of p53 was not significant. The IRSs of ras oncogene protein in the Recipe II-treated group, CTX-treated group and Runing Recipe II and CTX-treated group were lower than that in the normal saline control group respectively (P<0.05). CONCLUSION: Runing Recipe II can inhibit the growth of Ca761 breast cancer in mice by controlling the cell cycle of the transplanted tumor. This may be related to its effect on the gene expressions of p53 and ras in the tumor tissue.