The Preparation Of Mouse Model For Primary Hepatic Carcinoma And The Study Of Intervention For Huazhengsanjikeli

Primary liver cancer (Primary hepatic carcinoma, PHC) is the primary in the liver cells or intrahepatic bile duct cancer cells. Its incidence in China, second only to cancer, esophageal cancer. Currently, the high incidence of primary liver cancer, high mortality and its increasing trend year by year, has become an extremely serious international health problem. Clinical to liver pain, fatigue, poor appetite, abdominal distension, weight loss, fever, jaundice and other symptoms, to AFP, CEA, GGT, ALP and other abnormalities as features. Primary liver cancer is a common clinical cancer, the disease is difficult to detect early, the patient may be asymptomatic, patients with rapid progression of the disease in the late, highly malignant and prone to multiple organ metastasis, treatment more difficult, advanced liver cancer shortened survival in patients with natural features. The proportion of men and women with primary liver cancer is 2-5:1, clinical confirmed cases, mostly in patients with advanced, if not promptly treated, survival does not exceed six months. In medicine, "the liver volume, accumulation, Zheng Jia, jaundice, bulging, rock, swelling of the liver qi" and other liver disease attributable to modern medicine, the scope of application of Chinese medicine treatment can effectively control their primary liver cancer recurrence or metastasis, can improve the immunity of patients, can improve quality of life in patients with advanced, prolong life, and many other advantages, for the current comprehensive treatment of primary liver cancer an important component of treatment.1.Objective:Strains of mice with cancer, the subject of the passage with such complex modeling methods, preparation of primary liver cancer in mice animal models. Through the liver, spleen index, liver pathology and immunohistochemistry, gene expression, etc., as observed. Study of patients with primary liver particles scattered plot the intervention of a mouse model, in-depth study of its anticancer mechanism. For clinical treatment of hepatocellular carcinoma provides a theoretical basis, but also for the development of high efficacy, high-quality, high standards of traditional Chinese medicine preparation lay the foundation for liver cancer.2.Methods:Preparation of animal models of transplanted liver cancer H22, the animals were randomly divided into model group, group and disease for cantharidin product particles scattered low, medium and high dose group 5. Scattered plot of particle disease is low, medium and high dose group were administered the amount of 1.95g/kg, 3.9g/kg and 7.8g/kg; cantharidin dose group was 0.19g/kg; model group were given equal amount of sterile saline. Continuous administration, water supply for 14 days, 24 hours after the last administration, blood eyeballs, mice were killed, peeling the tumor, detection of liver, spleen index, and tumor tissue sections by HE staining, TUNEL, immunohistochemistry, RT-PCR and other methods to detect IL-2, Bcl-2, Bax, Caspase-9, Caspase-3 gene expression levels, and to plot the initial bulk of disease for the intervention of particles on the mechanism of liver cancer.3.ResultsThe results showed that:①disease for bulk product particles, high-dose group and cantharidin in liver, spleen index compared with model group were significantly different (P <0.01);②disease for bulk product particles, high-dose and cantharidin WBC Interleukin 2 (IL-2) expression was significantly higher than model group (P <0.05);③Compared with model group, positive particles in the bulk of disease, high dose group and cantharidin group of HE staining, tumor foci decreased, and with Normal organizational boundaries clearer.④TUNEL staining results: Mylabris group of disorders bulk product particles, high-dose group and model group with significant difference (P <0.01, n = 5))⑤immunohistochemical results: Mylabris group of patients scattered plot Particles, high-dose group and model group Caspase-9, Caspase-3, Bax has a strong positive expression, on the contrary there are weak Bcl-2 positive or negative.⑥RT-PCR, tissue Bcl-2, Bax mRNA expression was found in tumor tissue levels of Bcl-2 mRNA plot of scattered particles in the disease, the high dose group and cantharidin were significantly lower than model group (P <0.05), Bax mRNA level was significantly higher than the model group; of patients positive granules scattered Bcl-2 mRNAlow-dose group compared with the model slightly, Bax mRNA levels slightly higher compared with model group, but no significant difference (P> 0.05) .⑦Western-blot detection of Bcl-2, Bax, Caspase-9 and Caspase-3 expression can be seen that cantharidin group of disorders bulk product particles, high-dose group, Bax, Caspase-9, Caspase-3 protein expression was significantly higher than model group (P <0.05). In contrast, Bcl-2 protein was significantly lower than model group (P <0.05). 4.ConclusionCantharidin product group and disease for bulk grain, the high dose group can effectively inhibit tumor growth in situ carcinoma model, induced apoptosis of hepatoma cells. Cantharidin product group and disease for bulk grain, the high dose group may induce apoptosis of hepatoma cells by enhancing immunity, inhibition of Bcl-2 gene expression, while increasing the expression of Bax, followed by activation of Caspase-9, so Caspase-9 activation fragments increased, the final activation Apoptosis effector proteins Caspase-3, thus triggering a series of apoptosis cascade.