Effects Of Fei Ai Fang With Chemotherapy On The Treatment Of Non-small Cell Lung Cancer And Its Mechanism

Objective: To evaluate the effects of Fei Ai Fang with chemotherapy on the treatment of Non-small Cell Lung Cancer and to explore the primary mechanism of its main drug, Paris polyphylla, by observing its clincal impacts.Methods: This study was divided into two aspects: the clinical observation and its mechanism. In the clinical research, the treatment of “Fei Ai Fang” was deemed as an interventional factor. From July 2013 to May 2014, 66 patients who drove up the criterions of inclusion were enrolled and classified into two groups. Each group contained 33 cases. Patients who received the treatment of Fei Ai Fang with chemotherapy were defined as treatment group while the patients who received chemotherapy only were defined as control group. The chemotherapy consisted of TP regimen(including paclitaxel 135mg/m2 ivgtt d1 & cisplatin 75mg/m2 ivgtt d1). 21 days was 1 period. A week before chemotherapy, the treatment group began to take Fei Ai Fang for 14 days. After 3 cycles, we evaluated its comprehensive influences on the treatment of lung cancer patients by the criterions of tumor sizes, the living qualities, physical conditions, CEA, the phenomenon of medulla ossium suppression and the weight changes. In terms of the clinical results, we tried to find out the mechanism of anticancer activity of Fei Ai Fang. In research of its mechanism, we chose the large cell cancer line, NCI-H661 cells, as our subjects. WST-1 assay was used to examine the influences of different Chinese herbs in Fei Ai Fang on the cells, finding that the effectiveness of paris polyphylla was at the top. So the polyphyllin I(the main component of paris polyphylla) was selected as the interventional agent and we would study its mechanism. WST-1 assay was used for testing the IC50 of polyphyllin I and of the positive control drug(paclitaxel) on the cells. In the light of the results, the concentrations of the polyphyllin I to the experiment were determined. We divided the NCI-H661 cells into five groups at random, including the control group, positive control group(IC50 of the paclitaxel), 1, 5, 10μM polyphyllin I group. WST-1 assay was used to detect the growth rate of the cells and its growth curve was described. The cell cycle parameters and apoptotic rates were determined by means of FCM(Flow Cytometry) and TUNEL assay. The membrane permeability was measured via LDH kit. Western blotting was applied for assessing the expressions of caspase-3, caspase-8, caspase-9, cytochrome C, bim, bad, bax, bcl-2、p53 as well as cyclin D1 of the cells. With the intervention of different kinds of the blocking agents, the apoptotic rates were assessed by means of FCM(flow cytometry).Results: In the clinical research:(1) Comparsion of the general conditions: No statistic significance was found in the aspects of the ages, sexes, and TNM periods in two groups;(2) Comparsion of the tumor sizes: Based on the comprehensive explanations of imaging, the efficiency and stability rate of the patients in the treated group were higher than the patients in the control group in the aspect of tumor size;(3) Comparsion of the quality of lives: With the assession of the EORTC QLQ-LC43 scale, we found that the patients in two groups had improvements in the fields of physical functions, emotional functions, social functions, symptoms, the specific module of lung cancer and the overall survival qualities, compared with the former(p<0.05). Especially in the fields of physical functions, symptoms, the specific module of lung cancer and the overall survival qualities, patients in the treatment group improved more remarkable(p<0.05);(4) Comparison of physical improvement: KPS scores of the two groups were both higher than before and the score of the treatment group had more significance than the control one(p<0.05);(5) Comparison of tumor maker, CEA: After the treatment, the level of CEA in patients of two groups decreased obviously(p<0.01), and the treatment group’s decreased more significantly, presenting the obvious statistical difference(p<0.05);(6) Comparsion of the conditions of medulla ossium suppression: With the treatment, the condition of the medulla ossium suppression of the treatment group had a more notable alleviation than the control(p<0.05);(7) Comparsion of weight change: The patients in treatment group gained more weight than the control after treatment(p<0.05). In its mechanical investigation, the NCI-H661 cells’ behaviors of proliferation were depressed obviously with the treatment of polyphyllin I. The FCM(Flow cytometry) results indicated that the polyphyllin I could induce apoptosis but had specific effect on the cell cycle parameters. The morphology of the cells detected by TUNEL assay manifested the phenomenon of karyopyknosis and karyorrhexis. LDH kit helped us to learn that polyphyllin I could enhance the membrane permeability with a dose-dependence. Western blotting analysis showed that the levels of caspase-8、caspase-9、p53、bim、bcl-2 were decreased, the levels of bad、bax、cyclin D1、caspase-3 changed slightly and the level of cytochrome C seemed to have a small increase, however, without significance. In addition, the above impacts might be blocked by Pifithrin which could block the function of p53. On the other hand, 616452 and LY294002 seemed to be effective in blocking the functions of polyphyllin I, which, however, were weaker than Pifithrin. Taking the cell’s morphology into consideration, Pifithrin could protect cells from the exposure to polyphyllin I while 616452 and LY294002 could not.Conclusion: The therapy of Fei Ai Fang combined with chemotherapy will enhance the conditions of sufferers with NSCLC in many fields, such as tumor sizes, the quality of lives, physical conditions, tumor markers as well as the phenomenon of medulla ossium depression. Its main drug, the paris polyphylla, will depress the multiplication of NCI-H661 cells, whose mechanism may be related to the regulation of the functions of p53 and mitochondria as targets which will induce apoptosis. Therefore, there are more obvious advantages such as effect-enhancing and toxicity-reducing in the treatment of Fei Ai Fang with chemotherapy for the sufferers with NSCLC. It doesn’t only enlarge the application fields of Traditional Chinese Medicine but also provides clear evidences in order to promote its clinical extension in such field as tumor treatment.