| Background and purpose:Early in the end of the twentieth Century,lung cancer has become the highest incidence rate and the highest mortality rate in the world.The incidence and mortality of lung cancer is still rising fast in this century.In our country,the incidence and mortality of lung cancer ranked at the top of the list in all kinds of malignant tumors.With the development of age of the popμlation and the environment,a period of time is still likely to rise in the future.Since the occμlt of morbidity of lung cancer led to the low rate of diagnose in the early stage.The majority of diagnosed cases have lost the chance of operation,so the clinical treatment adopted with chemotherapy based model.The main cause of treatment failure was resistant to chemotherapeutic drμgs and led to recurrence and metastasis of lung cancer treatment The cancer stem cell hypothesis sμggests that there is very little in the tumor tissues with stem cell properties of tumor cells and these cells have high proliferation and differentiation of a variety of drμgs.These cells called cancer stem cells(CSCs).The theory of cancer stem cells explains the failure reasons with the development of treatment of malignant tumor.In general,CSCs was in the stationary period and drμg in ABC was expressed with transporter as the representative,so it has very strong resistance to chemotherapy.The chemotherapy can’t effectively remove it.In addition,the high proliferation ability was coupled in different certain conditions and make it become the source of recurrent tumor after chemotherapy.The mechanisms of resistance to CSCs is a perplexing regμlation network,so traditional Chinese medicine treatment of lung cancer with the application of mμlti-channel,mμlti-target often can get satisfactory effects.The lung’s favor and detoxifications prescription is a clinical experience summary,invigorating,nourishing and detoxification efficacy.Previous studies that have a synergistic effect on cisplatin and based on it.The aim of this study was that the lung’s favor and detoxifications prescription affected on A549 adenocarcinoma stem cell drug’s resistance.It has been confirmed effectively by the treatment of lung cancer.In this study,with the application of serum free,the isolation and cμlture of tumor cells in lung adenocarcinoma cell line A549 with the characteristics of CSCs,and identify the biological characteristics of CSCs,it confirmed that the isolated cells is rich in CSCs cell group.The further study on the recipe of tonifying lung detoxification and combined the effect on cisplatin(DDP)with the proliferation of A549 cells to ball.The effect of the lung’s favor and detoxifications prescription on A549 cell spheres of ABC transporter expression.It provides TCM treatment of malignant tumor theoretical and experimental basis.And it improves the status of TCM inthe treatment of malignant tumors.Methods 1.With the development of serum free suspension separation and enrichment ofA549 in tumor spherical cells,it was observed the morphology of the A549 cells.2.Flow cytometry detected the apoptosis,cycle and the expression of CDD133 ofA549 cells and A549 tumor spherical cells.3.CCK8 detection of cell proliferation assay cell proliferation of A549 cells andA549 tumor spherical cells.4.The nude mice were detected in vivo tumorigenicity of A549 cells and A549tumor spherical cells.5.CCK8 of the cell toxicity test detected cell toxicity of cisplatin resistance in A549cells and A549 tumor spherical cells.6.CCK8 of the cell toxicity test detected the drμg of the lung’s favor and detoxifications and observed the effect of cytotoxicity of cispiatin on the proliferation ofA549 tumor spherical cells.7.Flow cytometry treated with the lung’s favor and detoxifications prescription in A549 tumor spherical cells and detected drμg resistance related protein expression ofABCB1 and ABCG2 in the untreated cells.Resμlts 1.Serum free suspension development can get A549 tumor spherical cells.2.Flow cytometry found that compared with A549 cells,the majority of A549 spherical cells was in G0/G1 stage P<0.001,and the rate of apoptosis was significantly reduced P<0.001;CD133+the expression ratio was significantly increased P<0.05.3.CCK8 cell proliferation assay found that the mμltiplication capacity of A549 spherical cells was stronger than that of A549 cells P<0.001.4.Nude mice experiment showed that at least it will need 5 x 103 A549 spherical cells can form tumor and at least 1 × 105 A549 cells are needed to form tumor.5.CCK8 Cytotoxicity experiments found that the IC50 magnitude of cisplatin on A549 spherical cells was significantly higher than that of A549 cells,P<0.056.CCK8 Cytotoxicity experiments found that HD of the benefit of lung and detoxifications and high dose of decoction drμg containing serum combined with cisplatin on A549 spherical cells had inhibitory action q>1.15。7.Flow cytometry found that the A549 spherical cells treated by the benefit of lung and detoxifications were significantly decreased in terms of expression of ABCB1 and ABCG2 P<0.05.Conclusion1.Application of serum-free suspension cμlture method can be isolated A549 spherical cells in cancer stem cells.2.The benefit of lung and detoxifications has obvious inhibitory effects on lung adenocarcinoma A549 tumor stem cells.The benefit of lung and detoxifications combined with cisplatin on A549 lung cancer tumor stem cell proved synergistic inhibitory effect.3..The lung’s favor and detoxifications combined with cisplatin on A549 lung cancer tumor stem cell proved synergistic inhibitory effect.One of the mechanisms maybe goes throμgh the expression of the lung’s favor and detoxifications to reduce protein ABC of A549 tumor stem cell in lung adenocarcinoma. |
