The Change Of RhoA/ROCK Pathway Protein Expressed At Liver Subcutaneous Transplanted Tumors In H₂₂ Mice After Macrothele Raveni Toxin And Traditional Chinese Medicine Intervention

Objective: To compare Macrothele raveni Toxin and Anti-cancer Chinese medicine influence on Rho A/ROCK pathway protein expressed at Liver Subcutaneous Transplanted Tumors in H22 Mice and study the molecular mechanism.Methods: With reference to subcutaneous injection method, conduct subcutaneous vaccination of cell suspension of mice’s hepatoma carcinoma cell allocated in the concentration of 5x106pcs/ml on the neck of each KM mouse and make a model. After successful modeling, randomly divide the mice into three groups: model group(M), Anti-cancer Chinese medicine group(A) and Macrothele raveni toxins group(R), each group with 10 mice. Three days later, Each group of(M) were injected saline solution 0.3ml via caudal vein, and group of(R) were injected Macrothele raveni toxins 0.3ml with the concentration of 0.2μg/μl, While Mice in(A) group will be gavaged once a day(0.4ml/time) and dosing.All of the mice were injected 20 days, once a day. Measure the tumor size each 2 days. And draw the tumor growth curve. Observed microstructure of tumor tissue by HE staining method and using immunohistochemical to detect the expression of Rho A/ROCK Signaling pathways related proteins Rho A, ROCK and P21 in tumor tissues.Results:(1) Calculate growth rate of gross tumor volume according to measuring method of tumor volume: growth rate of(R) group is the slowest;(A) group takes the second place;(M) group is the fastest.(2) Display of HE staining: mice of(R) group: cells arrangement is sparse and irregular; tumor cell nucleus is in pycnosis; heterocyst can be discovered. Laminar cavitation denaturation and large areas of necrosis appear.(A) group: tumor cells arrangement is loose and out of order; necrosis area appears; cell nucleus is small and the destruction is inconspicuous; small amount of cavitation denaturation can be seen.(M) group mice: tumor cells arrangement is structured and compact; nucleus staining is dark and atypia is conspicuous; nucleoplasm is relatively large; border between parenchyma and mesenchyma is not clear; no conspicuous necrotic tissues.(3) Result of chemical staining of immune tissues: p21 positive expression rates of tumor tissues of mice in both(A) and(R) treatment groups are inconspicuously higher than that of(M) group, but Rho A and ROCK positive expressions of both groups are lower than that of(M) group(P<0.01).Conclusion: This study finds that spider venom from Macrothele raveni and anticancer traditional Chinese medicines have obvious inhibiting effect on the growth and proliferation of H22 hepatoma subcutaneous transplanted on mice. They lower the expression of Rho A and ROCK while the express of P21 protein is raised. The effect is shown most vividly in the group of spider venom from Macrothele raveni. Spider venom from Macrothele raveni and anticancer traditional Chinese medicines could restrain the proliferation and transferrence of tumor cells by adjusting the expression of relevant cell factors such as Rho A, ROCK and p21 in Rho A/ROCK signal pathway.