Study On ING5 Inhibition Of Epithelial-mesenchymal Transition In Breast Cancer

Objective:ING5 expression was studied in breast cancer tissues,We analyzed the correlation of ING5 with the parameters of clinical cases and aimed to study the expression of ING5 was correlated with the occurrence and development of breast cancer.Method:(1)We collected 68 cases of breast cancer tissue specimens,ING5 m RNA expression was detected by real-time PCR,and analyzed the correlation between it and the parameters of clinical pathology in breast cancer.At the same time,ING5 m RNA and protein expression were detected by real-time PCR and western blot in HMEC-1,MDA-MB-231 and MCF-7 cell line.(2)the cells line overexpressing ING5 was successfully constructed,invasion and metastasis activity was detected by Transwell chamber assay,apoptosis was detected by flow cytometry,and cell proliferation was detected by MTT assay.(3)We have successfully constructed ING5-overexpressing MDA-MB-231 and MCF-7 cell lines,PCR and western blot were performed to detect the expression of E-cadherin and N-cadherin m RNA and protein.Moreover,western blot was used to detect the expression of p-PI3K、 PI3K、p-Akt and Akt protein in MDA-MB-231 cells.We further use Image-Pro Plus 6.0 software to analyze the gray value of p-PI3 K and p-Akt protein,Results:(1)The expression of ING5 m RNA in breast cancer tissues was significantly lower than corresponding adjacent tissues,was correlated with tumor size,clinical stage in patients with breast cance.ING5 m RNA and protein in MDA-MB-231 and MCF-7 breast cancer cell line expression was significantly lower than in the expression in HMEC-1 cells.(2)Real-time PCR and western blot test results showed that the expression of ING5 m RNA and protein was significantly higher in the MDA-MB-231 and MCF-7 cells overexpressing ING5,as compared with the control group.These results suggest that overexpression ING5 of MDA-MB-231 and MCF-7 cells were successfully constructed.Test results Transwell chamber method showed that the migration and invasion ability in MDA-MB-231 cells and MCF-7 cells that overexpressing ING5 was significantly lower than the control group;Flow cytometry results showed significantly higher the number of apoptotic cells ING5 overexpressing MDA-MB-231 cells and MCF-7 cells,prompted ING5 induce breast cancer MDA-MB-231 and MCF-7 cell apoptosis.MTT assay test results showed that compared with the control group,the survival rate of MDA-MB-231 cell and MCF-7 cells overexpressing ING5 at each time point was significantly reduced;compared with the control group.(3)Compared with the control group,the expression of E-cadherin m RNA and the corresponding protein were significantly higher in MDA-MB-231 and MCF-7 cells which overexpressing ING5;and N-cadherin m RNA and the expression of the corresponding protein was significantly decreased;p-PI3 K and p-Akt protein expression was significantly reduced in ING5 overexpressing cells.the software results showed that the expression of p-PI3 K and p-Akt protein in ING5 overexpressing cells was significantly lower than the control group.The expression of PI3 K and Akt protein had no significant differences in the ING5 overexpressing cells.Conclusion:ING5 was abnormally low expression in breast cancer.When raised its expression can inhibit cell proliferation and induce apoptosis,invasion and migration ability of cells to make the drop.ING5 can upregulate the expression of E-cadherin and the expression of N-cadherin inhibited,showed that ING5 could inhibit EMT,and this inhibition was by inhibiting the phosphorylation of PI3 K and Akt,thereby inhibiting the PI3 K / Akt signaling pathway.