Research On The Role And Mechanism Of Glaucocalyxin A In Human Cervical Cancer Nude Transplantation Tumor Mice

BackgroundCervical cancer whose morbidity and mortality is at the forefront in developing countries, has been a threat on the health of women. Recently, with the improvement of traditional Chinese medicine extraction conditions, developing high efficient while low-toxic traditional Chinese medicine is the hottest research spot in the treatment of cervical cancer. Glaucocalyxin A which is one kind of extensive activity pharmacological two terpenoids, had been proved to inhibit a variety of tumor cell proliferation, however on cervical cancer there were no literature reports at home or abroad.ObjectiveTo build HeLa transplantation tumor model in nude mice, simulate the growth of cervical cancer in the human body environment, to explore the different concentration of Glaucocalyxin A in the body before and after cervical cancer inhibition, to explore different concentration of Glaucocalyxin A transplanted tumor cells before and after a prime role in the expression of p53 and Bax protein and gene level, in order to explore the mechanism of Glaucocalyxin A inhibition of cervical cancer and provide for the animal experiment basis for application in clinic.MethodsHeLa cells are injected subcutaneously into nude mice to build transplanted tumor model. When the tumor volume reached 0.22mm3,15 tumor-burdened rats are randomly divided into four groups before killing the mice, take blood from its eye to detect biochemical indices and tumor markers,in order to measure the changes of tumor markers, Transplanted tumor growth inhibition test, HE staining, TUNEL are used to observe the mice’s transplanted tumor morphological, histological patterns, cell apoptosis, immunohistochemistr, Western blot and RT-RCR, are used to analyze assay Glaucocalyxin A of different concentrations before and after transplantation tumor in the expression of P53 and Bax protein and mRNA expression.Results1. The tumor formation rate of nude mice was 100%.2. The results of biochemical index and tumor markers showed that there were no significant difference in Cr, BUN, ALT, AST, the WBC between the different Glaucocalyxin A concentration group and control group(P> 0.05), tumor marker CA125, CEA gradually reduced, with the incrsese of drug concentration of the Glaucocalyxin A, the comparative difference was statistically significant (P< 0.05).3. The transplanted tumor growth inhibition experiment showed that with the increase of the Glaucocalyxin A levels, tumor weight and volume reduce gradually, comparing differences between each group had significant statistical significance (P<0.01).4. The transplanted tumor pathological morphology observation showed that HeLa form was similar, before and after tumor tissue structure transplantation which indicates that the biological characteristics of approximate cervical cancer.5. TUNEL results showed, that the Glaucocalyxin A could induce the apoptosis of transplanted tumor cells, with the increase of drug concentration cell apoptosis index increased, comparing differences between each group had significant statistical significance (P<0.01).6. Immunohistochemistry, Western blotting and RT-PCR results showed that with the increase of the Glaucocalyxin A concentration, p53 Bax protein and mRNA expression gradually enhanced, the comparison between each group also had significant statistical significance (P< 0.01).Conclusions1. Glaucocalyxin A can restrain the growth of HeLa nude mice subcutaneous transplan- tation tumor and can induce apoptosis of transplanted tumor cell and can reduce the effect of tumor-burdened mouse tumor markers, and showed concentration dependence as well.2. Glaucocalyxin A may induce the expression of tumor suppressor gene P53 and apoptotic factor-Bax, thus inhibiting cervical cancer HeLa proliferation and promoting cell apoptosis.