| Objective:Previously,author research group found that Jinfu’an Decoction has an inhibition of lung cancer metastasis and development through impoving hemorheology.And then,they confirmed that JFA inhibits the development of A549 cell line and Lewis lung cancer of mice through vitro experiment and zoopery.The mechanism of it may be the expression of P120 ctn was treated.Kaiso is a member of Zine finger proteins family and also is a transcription factor of P120 ctn.Only P120 ctn can combine with Kaiso and then play a role in inhibiting cancer’s development and metastasis.Therefore,we sumised that JFA Decoction has an influence of inhibiting tumor development and metastasis through regulating P120 ctn and Kaiso.This research observes the influence of JFA Decoction on A549 tumor mice including act on anti-transfer and inhibiting tumor and quality of mice life.And also we observed the expression of P120 ctn,P120ctn 1A,P120ctn(PS288)and Kaiso.Based on these,we can discuss the mechanism of JFA Decoction on inhibiting tumor metastasis.MethodsUpon continuously passaging A549 cell line and tumor cell suspension was prepared.Sixty BALB/c-nu mice of SPF grade were randomly divided into 6 groups: high dose group,middle dose group,low dose group,positive control group(cisplatin group,model group,normal saline group and healthy control group).In addition to the healthy control group,the other 50 mice were inoculated in the right anterior axil with 0.2 ml suspension of A549 tumor cells to make the A549 tumor bearing mice model.It was fixed at 9: 00-11: 00 daily and observed the living conditions of the mice.The body weight,the number of deaths and the incidence of adverse events were recorded every 3rd.The drug was given for 21 days.After the experiment,all the mice were dislocated,the tumor and lung were removed by ophthalmic scissors,the tumor mass was measured and the tumor inhibition rate was calculated.The effects of JFA decoction and cisplatin on tumor inhibition and quality of life of mice were studied by means of body weight,tumor mass and food intake.The effects of JFA decoction on the phosphorylation of P120 ctn and its subtype 1A and related transcription factor Kaiso and P120 ctn serine 288 in A549 tumor-bearing mice were investigated by immunofluorescence,immunohistochemistry and real-time fluorescent quantitative PCR,and the expression of P120 ctn subtype 1A and P120 ctn serine 288 site were studied.So as to further explore the mechanism of A549 intervention in lung cancer metastasis.Results1.The effect of JFA decoction / cisplatin on the quality of life in mice was shows that: The increase in body mass and the intake of mice in each dose group of JFA decoction were better than those in the positive control group(P < 0.05).The tumor weight in the positive control group was lower than that in the JFA decoction group(P < 0.05).The number of death and adverse events in the positive control group were more than those in the positive control group.The mice in each dosage group and positive group showed obvious wasting and abnormal behavior.2.The results of indirect immunofluorescence assay showed that the a bnormal expression of P120 ctnntntntntn P120ctn subtype 1A,including dele tion,cytoplasmic heterotopic expression and nuclear expression,were dec reased in various dosage groups of JFA decoction compared with the model group.A relatively complete and bright green fluorescence appears on the cell membrane.The abnormal expression of P120 ctn in the positive group was lower than that in the model group.The phosphorylation of P120 ctn se rine 288 in the positive group and JFA decoction group was lower than tha t in the model group.In the positive group,the expression of Kaiso prot ein was weak and diffuse in the cytoplasm,and the expression of green fl uorescence in the cytoplasm of each dose group of JFA decoction was signi ficantly lower than that in the model group,and almost no expression was found in the positive group.3.The abnormal expression of P120 ctn and its subtype 1 in JFA decoct ion group was lower than that in model group and positive group in immuno histochemical experiment,and the amount of abnormal expression in JFA decoction was the least.The positive expression of P120 ctnnnntntntntntntn tntntntntntntntntnh288 in the model group was the highest in the model gr oup,and decreased in all the dosage groups of JFA decoction.The differe nce between the model group and the model group was statistically signifi cant(P < 0.05),and the expression of P120 ctnnntntntntntnc288)was the least in the positive group.The difference between the model group and t he model group was statistically significant(P < 0.05).Kaiso was positi ve in both the model group and the low dose group,but the score score of the low dose group was lower than that of the model group.The positive expression of Kaiso in the middle dose group of JFA decoction was found i n the model group.Each dose group of traditional Chinese medicine was th e least,There were significant differences in the score and positive rat e of the model group(P < 0.05).The positive expression of Kaiso in the positive group was the least in each treatment group,and the difference in the model group was statistically significant(P < 0.05).4.The results of real-time fluorescence quantitative PCR showed that the expression of P120 ctn mRNA and the expression of Kaiso mRNA in different dosage groups of Jinfuan decoction were lower than those in the model group,and the mRNA expression in the medium dose group of P120 ctn was the least.The expression of mRNA in the low dose group of Jinfuan decoction was the same as that of P120 ctn mRNA in the traditional Chinese medicine group.There was significant difference in the expression of P120 ctn in the model group compared with the model group(P < 0.05)and the expression of P120 ctn in the positive group was slightly higher than that in the model group.The difference was not statistically significant(P > 0.05).However,the expression of Kaiso decreased significantly in the positive group(P < 0.05).Conclusion1.In animal experiments,JFA decoction has the effect of controlling tumor,although the effect of inhibiting tumor is not as good as that of cisplatin,it can improve the food intake and nutritional status of mice,thus improve the quality of life of mice.2.JFA Tang may interfere with the abnormal expression of P120 ctn and its subtype 1.Reduce the phosphorylation of P120 ctn serine 288 site and the cytoplasmic expression of P120 ctn transcription factor Kaiso,thus inhibit the development and metastasis of lung cancer. |
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