| BACKGROUND:Primary liver cancer (PLC) has become the second cause of malignant tumors, which is one of the most common malignant tumor. Statistics showed that the new cases about 45 percent of liver cancer cases all over the world each year are in the mainland of our country, the incidence of liver cancer mortality was increased by 41.7% in the past 20 years. But in recent years, the study showed that treatment of traditional Chinese medicine with conventional Western medicine for advanced primary liver cancer has significant improvement in clinical symptoms and signs and improve quality of life, prolong survival time. In recent years, many Chinese herbal medicine have been developed. The components filtered and purified from Chinese herbal medicine, such as traditional Chinese medicine polysaccharide, have been used in anti-tumor experiments. Some progress has been made. It has provided a way for anticancer medicine.Trametes robiniophila is medicinal fungi in aged Chinese Trametes tree (Trametes robiniophila Murr). Trametes robiniophila obtained by hot water contains a variety of organic ingredients and more than ten kinds of mineral material elements. The main active ingredient for anticancer protein is polysaccharide (PS-T). Now there are its products Jinke Huaier Granule (National Class One drug) for clinical application. Modern study reports show Huaier Granule clinically has been widely used in a variety of tumor treatment, and good effects have been achieved. 1. For digestive system tumors: 5years survival rate of gastric cancer has significantly increased;united chemotherapy drug can improve immune function in colorectal cancer patients, reduce the toxicity of chemotherapy and improve the hematopoietic function;2. For respiratory system cancer: Huaier can significantly improve the non-small cell lung cancer patient's immune function;3. For blood tumor: the treatment of chronic myelogenous leukemia have a total efficiency up to 80%;4. other cancer: Trametes robiniophila also has good effect for Breast Cancer. In explaining its anti-tumor mechanism, many experiments also have been carried out. Trametes robiniophila basic role in liver cancer experiments show that in vitro Trametes robiniophila has direct use on liver cancer cells and induces apoptosis. This experiment will be on this basis, further research Trametes robiniophilainduced apoptosis of hepatoma cells. OBJECTIVE:1.In vitro observing growth inhibition of human hepatocellular carcinoma SMMC-7721 cell lines on Trametes robiniophila;2. To observe inhibition of hepatocell ular carcinoma cells in mice transplanted tumor and to explore its mechanism on Trametes robiniophila.METHODS:1. MTT assay the inhibitionrate of in hepatoma cells SMMC-7721 cell line by Trametes robiniophila in vitro.2. To observe inhibition and induction of apoptosis of mouse transplanted tum or on Trametes robiniophila.Subcutaneous injection of H22 hepatocarcinoma cells to mice, creating a model tumor-bearing mice , they were divided into A group (control group), B group (5-Fu group), C group (Group Trametes robiniophila) and D group (Trametes robiniop hila+5-Fu group), tumor weight was weighted at the end of treatment, calculating t he inhibition rate, HE staining is to observe pathological changes in tumor tissue; apoptosis in tumor tissue in each group rate by TUNEL assay;transplantation tum or related apoptosis protein Bcl-2, Bax expression, to explore its possible mechanis m of induction of apoptosis by immunohistochemical method.RESULTS:1.Inhibition of cell proliferation in vitro experiments: hepatocellular carcinoma SMMC-7721 cells were significantly inhibited by Trametes robiniophila and the concentration increases with the dose effect relationship, 10 mg/ml Trametes rob iniophila group and 5 mg/ml Trametes robiniophila group 48 h inhibition rate reached 65% and 55% respectively.2. In vivo tumor growth inhibition experiment: the weight of tumor-bearing mice in A group was significantly is greater than B group (5-Fu group), C group (T rametes robiniophila) and D group (Trametes robiniophila +5-Fu group) (P<0.05, P<0.05, P <0.05), B, C, D Group inhibitory rates were 31.2% and 27.1%, 47.5% respectively, suggesting that Trametes robiniophila in the body can inhibit the growth of tumor;and the inhibition rate in D group was significantly higher than B, C group, (P<0.05), prompting Trametes robiniophila United 5-Fu drug, 5-Fu can enhance the effect of inhibiting tumor . 3.Tumor cell apoptosis rate by Tunel, detection showed that the apoptosis rate high to low as follows: Trametes robiniophila United 5-Fu group, Trametes robiniophila group, 5-Fu group and the control group, the difference has statistical significance (P<0.05).4. Related apoptosis protein Bcl-2 determination, immuneohistochemical results showed that the control group of Bcl-2 positive expression rate is 43.02±2.05%, is significantly higher than 5-Fu group, Trametes robiniophila group, Trametes robiniophila United 5-Fu group.The positive expression rate, the difference has statisti cal significance (P<0.05), showed that clearance of Trametes robiniophila paste can be reduced tumor-bearing mice tumor cell Bcl-2 protein expression.5.Related to the determination of apoptotic protein Bax, immunohistochemical results showed that: Trametes robiniophila united 5-Fu group Bax positive expression rate is 87.88±9.72%, which was higher than 5-Fu group, Trametes robiniophila group, (P <0.05);difference between the control group and Trametes robiniophila Group has no statistically significant (P>0.05). It showed that Trametes robiniophila united 5-Fu can enhance the expression of Bax protein.CONCLUSIONS:1.Trametes robiniophila in vitro have inhibited growth of hepatoma SMMC-7721 cells2.Trametes robiniophila in vivo can inhibite tumor growth in mice by induction of apoptosis;3.Trametes robiniophila can reduce Bcl-2 gene protein expression, united 5-Fu, Bax gene protein expression increase apparently, Trametes robiniophila may inhibit tumor growth in tumor-bearing mice by inducing apoptosis of transplanted tumor.
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