The Study Of The PI3K-Akt Signaling Pathways Regulate The Process Of Invasion And Metastasis In Human Breast Cancer Cell Mediated By The Osteopontin

Breast cancer is the most serious cancer disease does harm to the global women health.With diagnosing the breast cancer earlier,and as soon as possible to take surgical treatment methods can prolong patients’ life effectively or cure the disease.However,due to earlier symptoms of breast cancer is not easy to be perceived by patients,so there are a lot of patients at the time of diagnosis,the malignant degree of breast cancer has been aggravating,and the cancer cells had shifted.At present,the treatments of metastatic cancer cells have poor effect.In the patients with cancer metastasis,their survival rate are very low.Therefore,the research of the treatments to inhibiting cancer metastasis have much attention.TCGA database is using the method of genome sequencing and bioinformatics methods to classify,summarize and integrated the data of the cancer gene expression,the gene mutation and the clinical.So as to improve the ability of the diagnosis,treatments and prevention of cancer.To make the researchers to understand the genetic basis of cancer better.To provide the basis for the study and treatment of cancer.PI3K is playing an important role in regulating the cell growth,proliferation and differentiation process.But in breast cancer cells,because of the high amount of the expression of PI3 K,it’s occurring excessive activation of Akt,and it often lead the PI3K-Akt signaling pathways to imbalance.This has closely related with the breast cancer occurrence and developmen.Osteopontin(OPN)is a kind of phosphorylated glycoprotein.OPN can play a role in cells,and can also secrete to develop its physiological function outside the cells.In the breast cancer,the OPN has prompted many kinase activation,so as to promote the process of invasion and metastasis of cancer cells.Studies have shown that the PI3K-Akt signaling pathway have the relationship with the OPN expression.Therefore,this study has selected breast cancer as experiment object to investigate whether PI3K-Akt signaling pathways are played an important role in the regulation of the expression of OPN and have the impact on the process of invasion and metastasis of breast cancer.It has important practical significance for the prevention and treatment of breast cancer invasion and metastasis of tumor proliferation.Firstly,we obtain the gene expression data and clinical data of the patients with breast cancer from the TCGA,and analyze the PIK3 CA and SPP1 gene graphic in cancer tissue and normal tissue adjacent to carcinoma,cancers with different levels and different stages of cancers.Then we analyze the expression of these genes influence on patient surial and analyzes the relationship between the expression of these genes.According to the results of the analysis,we think that the over-expression of the PI3 K can make the PI3K-Akt signaling pathways excessive activation,and then make the OPN over-expression and promoting the breast cancer invasion and metastasis.PI3K-Akt signaling pathways play an important role in regulating for the expression of OPN in breast cancer.In order to prove these points,we select the normal mammary gland cell lines,low metastatic breast cancer cell lines,high metastatic breast cancer cell lines and obtain the relevant clinical samples from hospital as the experimental materials for experiment.First of all,we use real-time fluorescent quantitative PCR,Western Blot,cell climbing immunohistochemical experiment and immunohistochemical experiment to determine the differences of the gene and protein expression level of the PI3 K,Akt and OPN in the different types of breast cancer cells and tissues,and in the normal breast cell and normal tissue adjacent to carcinoma.And then,we chose a specific inhibitors for PI3 K catalytic subunit p110α,it’s called A66.To deal with the MCF-7 and MDA-MB-231 cells respectively.We use real-time fluorescent quantitative PCR,enzyme linked immunosorbent assay,Western Blot and cell climbing immunohistochemical experiment to detect the change of the gene and protein expression level of OPN after the two types of cells were processed by the inhibitors.We use the MTT experiment and cell wound healing experiment to testing the change of the ability of proliferation and migration of the two types of cell after processing by the inhibitors.We use Transwell cell invasion experiment and soft AGAR cloning formation experiment to examining the change of the ability of invasion and the un-anchoring growth of the MDA-MB-231 cells after processing by the inhibitors.According to the results of these experiments,it’s proved that the PI3 K is over-expression in the breast cancer,and then promote the Akt phosphorylation,let the PI3K-Akt signaling pathways excessive activation and the gene of OPN over-expression.The genes expression of PI3 K and OPN and the generation of p-Akt will rise with the increase of breast malignant degree.It’s shown that the PI3K-Akt signaling pathways have played an important role in the regulation of the expression of OPN,and have the influence on the process of invasion and metastasis of breast cancer.This study provides a new idea for the researches of the mechanism of the breast cancer cells invasion and metastasis,and offers a new basic data for the treatments of the patients with breast cancer.