| Lung cancer is one of the most common human malignancies,It has been ranked the top cause of death of all malignant tumors in many cities in China.The majority of our areas of lung cancer morbidity and mortality in the past 20 to 30 years grew rapidly,and will continue to maintain this growth trend,Lung cancer has become one of the most important diseases threating the health of our people.Chemotherapy is the major mean to treat the advanced cancer,the shortcoming of chemotherapy is to produce great damage not only to the tumor cells,but also the normal cells,especially to blood cells,bone marrow cells,and so on.Cisplatin(DDP) has stronger role in the antitumor drugs,it has significant clinical effect to the various solid tumors,but its serious side effects impact the quality of life of patients greatly.It is extremely important to find some natural treatment of cancer drugs with low toxicity,and fewer side effects and have a certain effect,or use them to increase the sensitivity of chemotherapy drugs.These substances have two main sources currently: one is plant secondary metabolites in the plant foods,and the other is the anti-cancer substances in Chinese herbal medicine.Gegen isoflavone is the ingredient extracted of dried legumes kudzu root.In recent years,researchers have found that its anti-cancer role,but its anti-cancer activity is lower than puerarin crude extracts.Our pueraria crude extract is the main biologically active ingredients group from the drug edible plants puerarin extracted with alcohol extraction,compared to standard of pure puerarin,its extraction method is relatively simple and retain more of the active ingredients,The purpose of this study is to discuss these materials can enhance chemotherapy with cisplatin or not.This experiment use of human Small Cell Lung Cancer H446 cells as target cells, using MTT assay,flow cytometry,immunohistochemistry to explore cisplatin alone or combined with pueraria crude extract and standard of pure puerarin changing the cell proliferation of Small Cell Lung Cancer H446 cells.To observe the role of the puerarin puerarin extracts and different concentrations of cisplatin alone or jointly by cell proliferation and apoptosis aider the change,and provide experimental basis of pueraria crude extract and standard of pure puerarin sensitizing effect to the chemotherapy cisplatin.ObjectiveThe purpose of this study is to study the different effects of standard of pure puerarin(SP),pueraria crude extract(CP) of low concentrations on the in vitro growth of H446 cells of Human Small Cell Lung Cancer when employed independently or jointly with DDP.By measuring the relationships between cell growth,cell cycle and their relative genetic expressions,I further discuss the related mechanism of tumor cell apoptosis and provide theoretical evidences for the making of rational clinical treatment plan with integrated chemotherapy and the seeking of optimum curative effect.MethodsIn this experiment,8Oμg/mlSP,CP was employed independently or jointly with DDP in the H446 cell lines of Human Small Cell Lung Cancer.Methabenzthiazuron (MTT) colorimetry is adopted to examine the inhibition effect of drugs on cell growth. The rate of cell apoptosis is examined by flow cytometry method.The expression of bcl-2 and bax are investigated by immunohistochemical techniques. Results1.SP,CP of five different concentrations(40μg/ml,80μg/ml,160μg/ml,320μg/ml and 640μg/ml) can inhibit the growth of cells in various degrees after they were added to in vitro growing H446 cell lines for 24 h.The respective inhibition ratios differ markedly due to difference in concentration(P＜0.05).They were 0.09%,2.03%, 10.90%,17.03%,22.64%for SP;1.70%,3.49%,12.17%,19.29%,27.03%for CP.2.When SP,CP was employed jointly with DDP in H446 cells,the effect was stronger than When DDP is used independently(P＜0.05),showing concentration dependency;CP is stronger than SP when used independently or jointly.3.When the SP,CP was jointly used with different concentration DDP for 24 hours. The apoptosis inhibition ratios were 6.1%,17.5%,24.4%,26.6%for SP and 15.6%, 21.2%,25.4%,37.7%for CP.the effect was stronger than When DDP is used independently.4.When testing bcl-2 and bax by immunohistochemical techniques,their express rates changed markedly with the increase of concentration(P＜0.05).The expressive rates of bcl-2 are decreased and the expressive rates of bax are increased.Conclusion1.SP,CP has no effect on H446 cell growth when the concentration is below 80μg/ml.2.SP,CP can enhance DDP on inhibiting the H446 cell growth in Human Small Cell Lung Cancer.3.SP,CP can enhance DDP block H446 in the G0/G1 phase,induced cell differentiation.4.The inducement of SP,CP can inhibit bcl-2/bax expression.This may attribute to one of the molecule mechanisms that SP,CP can resist the multiplication of cells and accelerate cell apoptosis.SP,CP have an enhanced sensitivity effect on DDP.5.CP combined with cisplatin has stronger role than SP in cell growth inhibition,cell differentiation. |
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