Thechangeof P53/P21pathwayproteinexpressedat Liver Subcutaneous Transplanted Tumorsin H₂₂ Miceafter Macrothele Raveni Toxinandtraditional Chinesemedicine

Objective:To compare intervening effect of Macrothele raveni toxin and Traditional Chinese medicine to the p53/p21 signal pathways,the relevant protein factors of subcutaneoustransplantationtumorofmice H22,and relevantmolecularmechanism.Methods: The H22 mice hepatic tumor model was constructed by a subcutaneous grafting method in 30 KM mice. After the model was successfully constructed, mice were randomly divided into 3 groups: a model control group, a Macrothele raveni toxin group, and a Traditional Chinese medicine group.Three days later, each group receive the intervention of Macrothele raveni toxin and Traditional Chinese medicine.The model control group receive an intravenous injection in the tail of 0.3ml saline. The Macrothele raveni toxin group receive an intravenous injection in the tail of 0.3 ml Macrothele raveni toxin at the concentration of 0.2 ug/ul. The Traditional Chinese medicine group were fed mice0.4ml, once a day, were administered 20 days. Fasting the mice overnight after the experiment, then sampling the tumor tissue based on mice and calculate the inhibitory rate The hematoxylin and eosin(H&E) staining method was used to analyze the microstructure of hepatic tumor tissue and immunohistochemistry was used to detect the differential expression of mutant type P53, P21, Bax, and Survivin protein levels in tumor tissue.Real-time PCR was also used to detect the expression level of genes encoding tumor wild type P53, P21, Bax, and Survivin m RNA.Results:The Calculation of the inhibitory rate shows that the Macrothele raveni toxin group is higher than the Traditional Chinese medicine group;①H&E staining demonstrated the following results:(1) the group administered Macrothele raveni toxin had a large necrotic area in the tumor,(2) the group administered Traditional Chinese medicine had shrinkage of the cell nucleus in tumors and a necrotic area was observed,and( 3)the control group had no obvious tissue necrosis and abundant blood vessels could be seen.② Immunohistochemical detection reveals that thepositive expression of p21 and Baxproteinsforgroupof Macrotheleravenitoxin,groupof Traditional Chinese medicine display obvious rise when compared with that of the model group, on the contrary,mutantp53 and Survivinproteinsshowsevidentdecreasewhencomparedwith that of the model group. ③ Real-time PCR detection reveals that for the group of Macrothele raveni toxin, group of Traditional Chinese medicine, the expression of wild-type P53, P21 and Bax m RNA increase obviously when compared with that of modelgroupwhiletheexpressionof Survivinm RNAislowerthanthatofmodelgroup.Conclusion: Three intervention means as Macrothele raveni toxin, Traditional Chinese medicine could effectively improve the content of P21 and Bax-associated proteins and m RNAand wild-type P53 m RNAand reduce the content of mutant P53 and Survivin proteinsandm RNAvia P53/P21 signalpathtoexertthefunctionofrestrainingthetumor proliferationof H22 hepatomacell line.With regardstotheregulation effect,theeffect of Macrotheleravenitoxinisthemostobvious.