Study Of The Anti-tumor Activity Of Paris Polyphlla And The Effect Of Traditional Chinese Medicine Combimed With Fluorouracil Drugs

[Purpose]To research the safe dose of Paris polyphlla ethanol extract(PPE)through the acute toxicity of ICR mice.To investigate the application of PPE in the comprehensive treatment of cancer,and to find its antitumor effect and mechanism of PPE in vitro.To investigate the relationship between the preoperative level of serum tumor markers and cancer-related venous thromboembolism in patients with gastric cancer.To preliminary discuss the effect of fluorouracil application in clinical using chemotherapy combination with Traditional Chinese Medicine for gastric cancer.[Method]1.The acute toxicity experiment was carried out using ICR mice.2.MTT and cell cloning methods was used to determine the growth inhibition of the MKN-45 cells by PPE or PPE combined with radiotherapy.PI staining was applied to test cell cycle of the MKN-45 cells on PPE or PPE combined with radiotherapy.Western-blot was used to detect apoptotic protein levels of PPE.RT-PCR was used to detect changes of TS mRNA levels of the PPE.3.1005 patients with gastric cancer who underwent curative resection were finally enrolled in this study.Serum tumor markers including CEA,CA19-9,CA242,CA72-4,CA125 were tested within one week before curative surgery and also after 6 cycles of chemotherapy.The sensitivity,specificity,positive predictive values,and negative predictive values of five biomarkers and the combination group were calculated respectively.From that we find 270 patients need adjuvant chemotherapy.TS mRNA level and tumor markers was tested before and after chemotherapy.Comparing serum makers between the two groups,one year relapse-free survival,and peripheral blood TS mRNA level changes before and after chemotherapy.[Result]1.The LD50 value of ICR mice p.o.with the PPE was 12mg/ml.2.The growth inhibiton of human gastric cancer cells MKN-45 by PPE was manifested in a time and dose dependent manner.Cell cycle results suggestted that PPE could block cell in S period and G2/M period without or with radiotherapy.Furthermore,PPE could improve the apoptosis of MKN-45 cells after 48 hours treament.Besides,PPE could downregulate the expression level of bcl-2 protein and upregulate the expression level of Bax protein.The PPE could also reduce half levels of TS mRNA in MKN-45 cell line.3.Cancer-related venous thromboembolism was found in 540(53.73%)patients.The increase of each cancer marker was remarkably related with cancer-related venous thromboembolism.The more abnormal serum tumor markers come out,the higher the risk of positive cancer-related venous thromboembolism could be.The possibility of positive cancer-related venous thromboembolism would be 2.9 times as much as a normal patient as any of the tumor markers increases.With the increase of any 4 of the tumor markers can make the risk 16.4 times.The peripheral blood tumor markers increased by more than 20%was found in 27(19.3%)and in 34(26.2%)patients after six-cycles treatment in traditional Chinese medicine combination group and in mono-chemotherapy group,respectively(P = 0.178).In addition,relapse was found in 31(22.1%)and in 43(33.1%)patients in one year,respectively(P=0.044).No significant difference of peripheral blood TS mRNA before chemotherapy was observed between two groups(Comb vs.Mono:1.31 vs.1.28,P=0.93).After six cycles' treatment,the peripheral blood TS mRNA level increased in mono-chemotherapy group(before vs.after:1.28 vs.1.42,P=0.43)and decreased in combination group(before vs.after:1.31 vs.0.98,P=0.12).However,no significant difference was observed.[Conclusion]1.The toxicity of PPE may be caused by the change of white blood cell.Thus,attention should be paid to hematological test results during clinical application 2.The PPE revealed significant anti-tumor effect in vitro,the mechanism may be related to affecting the cell cycle,inducing cell apoptosis through down-regulation the protein expression level of Bcl-2 and up-regulation the level of Bax,sensitizing chemotherapy drugs through the improving of TS mRNA level.Besides,the combination of PPE and radiation can lead to the releasing of ROS which is relevant to the blockage of the cell cycle and apoptosis of tumor cells.3.The status of postoperative cancer-related venous thromboembolism can be predicted from preoperative serum tumor markers levels.For those patients with a high risk of positive cancer-related venous thromboembolism,more aggressive treatment should be arranged.4.Compared with chemotherapy alone,chinese traditional medicine in combination with chemotherapy can significantly reduce the risk of tumor recurrence in stage ? gastric patients who received adjuvant chemotherapy after surgery.The mechanism might correlated with reduced TS mRNA expression caused by chinese traditional medicine which associated with the sensitivity of fluorouracil.