Preliminary Study On The Mechanism Of CCR5 Receptor Antagonist Promoting Apoptosis Of A549 Cells In Non-small Cell Lung Cancer

BackgroundAt present,malignant tumors are the main causes of global morbidity and mortality.In 2012,lung cancer accounted for 20% of all deaths and 8 million 200 thousand of all deaths caused by malignant tumors,accounting for the highest death rate of all cancers.By 2015,the number of new cancers around the world is 14 million 100 thousand,and it is estimated that the number of newly increased patients will increase by 70% in the next 20 years.Due to the growth of age,poor living habits,environmental pollution and the low cure rate of traditional means,China has become a major country of cancer and death.2 million 810 thousand cases died,about 75% of the patients had progressed to advanced,and the survival of not more than 5 years.At present,the effect of treating non-small cell lung cancer is not ideal.The malignant proliferation of cancer cells and metastasis of cancer foci bring great difficulties to clinical treatment.The tumor microenvironment has attracted more and more attention.Many studies have reported that there are a lot of chemokines,and the interaction between interstitial cells and cancer cells induced by the internal environment,resulting in the immune inflammatory response,has promoted the invasion of cancer cells.Recent studies showed that chemokine receptor CCR5 binds to chemokine CCL5 and promotes cancer cell invasion and metastasis,although there are some reports of CCL/CCR and apoptosis of tumor cells,and CCL cells and an important tumor suppressor gene of P53 signal pathway related,while inhibition of CCR5 receptor can promote apoptosis the tumor cells has not been reported.Therefore,the study of the relationship between the CCR5 receptor inhibitor and the apoptosis of non-small cell lung cancer cells can provide a good basis for the clinical treatment of lung cancer.ObjectiveObjective to study the mechanism of CCR5 receptor antagonists in promoting the apoptosis of non-small cell lung cancer A549,and to provide new research ideas for the prevention and cure of non-small cell cancer of lung,and to offer a theoretical basis for helping develop new targeted anti-tumor drugs.Methods1.The use of CCR5 receptor antagonist nifeviroc,at the same time and concentration,in non small cell lung cancer cell A549,non-small cell lung carcinomaH520 cells,normal bronchial epithelial cells HBE,breast cancer cells MCF-7,HePG2 cells,and the proliferation of MTT was detected by cell inhibition rate.2.Using CCR5 receptor antagonist nifeviroc,at the same time and concentration,to impact cells which are different kinds,and the flow cytometry is used to test the rate of apoptosis in different kinds of cells.3.Flow cytometry was used to detect the expression of CCR5 receptor on the surface of the different species.4.By using Western blotting(Western Blot),a preliminary inquiry nifeviroc mechanism to promote the apoptosis of A549 cells.Results1.In nifeviroc effect,using MTT method to detect with nifeviroc concentration and the prolongation of time,A549 and MCF cells have greater growth inhibition and HBE inhibition rate of HePG2 is relatively low,while H520 cells were almost not affected.2.In nifeviroc effect,using flow cytometry to detect the apoptosis of different cell types,the rate of apoptosis of A549 cells,which was the highest,MCF cells,HBE and apoptosis rate of HePG2 cells was low,almost no apoptosis of H520 cells.3.By flow cytometry,the expression of CCR5 receptor on the surface of cells from high to low was A549 cells,MCF cells,HePG2 cells,HBE cells and H520 cells.4.In nifeviroc after using Western Blot technology to detect the inhibition of PI3K/Akt signaling pathway,reduce the energy of protein Akt activity(m TOR),rapamycin receptor expression,tumor suppressor gene P53 protein content increased,cyclin Cyclin decreased D1 content.Conclusion1.Flow cytometry was used to detect the expression of A549 cell surface receptor CCR5 was the highest,A549 cell apoptosis after nifeviroc effect was the highest,suggesting that the rate of apoptosis was correlated with CCR5 expression.2.Nifeviroc can promote A549 cell tumor suppressor gene P53 expression increased,and the phenomenon contributes to cancer cells apoptosis.3.In nifeviroc role,an important signal transduction pathways of PI3K/Akt cells in A549 cells was blocked,the physiological function is inhibited,causing apoptosis.