| Background Primary liver cancer is a common malignant tumor in China. It is the second cause which leads to death, especially in countryside. At present its prevalence has an increase tendency. Now the surgical operation is still the first choice, however, the effect isn't very satisfying because the rate of postoperation relapse is very high. On the other hand, radiotherapy and chemotherapy can't kill all tumor cells, and they are easy to impair the normal tissues. So we need to seek new treatments. Recently biological therapy has become the fouth model in treating tumors which mainly includes inhibiting cell proliferation and promoting cell apoptosis. Anyway, anti-sense gene therapy plays an important role because it is easy to operate and control. At present it means anti-sense oligonucleotides technology. Because the pathogenesis of hepatocellular cancer is characterized as multi-factor, multi-stage, multi-gene and multi-mutation.The choice of exact targeting gene is very crucial. Altieri et al first clones the gene of Survivin in 1997. The protein is the strongest apoptosis inhibitor at present called survivin protein. Its biological effects includ: â‘ inhibit cell apopt, â‘¡regulate cell proliferation, â‘¢induce angiogenesis. Survivin can express in embryonic tissues, but undetectable in differentiated adult tissues It can be detected in many human tumors. And it is the fouth of over 100 tumor-specific genes. Survivin can't express in normal tissues, so it is regarded as the ideal anti-sense therapeutic target. At present the research on the relationship of Survivin and the pathogenesis of liver cancer is few. Especially there is no report about the relationgship about Survivin, Caspase-9 and its effect in inhibiting liver cell apoptosis and angiogenesis. Anti-sense Survivin gene therapy in the field of liver cancer is very limited. This study proves that Survivin can stably express in liver cancer to inhibit cell apoptosis, and induce angiogenesis. It can function as a kind of poor prognosis index. We design anti-sense oligonucleotides sequences targeting Survivin mRNA translation initiation coden which can transfect SMMC-7721 cells to study the proliferation and apoptosis of liver cancer. This can open a new way for the comprehensive evaluation and biological therapy.Objective 1. We study the expression of Survivin and Caspase-9 in primary live cancer and their effect on cell apoptosis and angiogenesis, and discuss their effect on the pathogenesis of liver caner and biological action. Finally, we offer a new index for liver cancer comprehensive evaluation. 2.Anti-sense oligonucleotides sequences targeting Survivin can transfect SMMC-7721 with cation liposome to examine its effect on liver cancer cell proliferation and apoptosis, this offer a new way for biological therapy of liver cancer finally.Materials and Methods Materials 1.Target (1) tissues specimen: 42 primary liver cancer tissues were obtained by the surgical resection from the department of general surgery, the first hospital of Jilin University from1998 to 2000: 33 male and 9 female, 16 (<60yr) and 26 (>0yr). According to Edmondson Grade Criteria, Grade I is 8 cases, Grade II is 10, Grade III is 16, Grade IV is 8. The diameter of tumor (<5cm) is 16 cases, >cm is 26.Metastasis tendency (lymph node metastasis, portal vein cancer thrombosis, distant metastasis, the tumor amounts >2, satellite node around cancer) is 19 cases. 42 nontumor tissues were those which were 2cm away from the lesion, including 28 cirrotic tissues, 10 hepatitis tissues and 4 normal liver tissues. 12 normal liver tissues were obtained by autopsy from Faculty of medicine of Jilin University. All tissues were fixed in 10% formalin for slices staining. (2) Cell lines: SMMC-7721 is purchased from Shanghai cell research institute of Chinese Academy of science. 2.Equipment: Microtome, Incubator, Hyperclean Working platform, Ultraviolet Spectrophotometer, PCR Flycyte Meter, Enzyme Labelling Meter, Transmission Electron Mic...
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