| Hepatocellular carcinoma (HCC) is the most common liver malignancy. The main treatments are resection, embolism chemical therapy or ablation, but the 5 years survival rate of HCC patients is lower than 25%. The reason of bad prognosis is mostly related with the recurrence and metastasis after treatment. This study aimed to determine the differences of prognosis between these treatments and the changes of the cytobiological characteristics between the primary and the recurrent tumor.Objectives:1. To figure out the difference of survival time of the tested mice which were treated by resection, percutaneous ethanol injection (PEI), Argon-Helium cryotherapy or untreated.2. To study the different cytobiological characteristics of the tumors after these three kinds of treatments by detecting the expression of residual tumors on p27, bcl-2 and survivin.3. To evaluate the differences of the cytobiological characteristics between the recurrence tumors and the primary tumors of the tested mice by detecting the expression of p27 , bcl-2 and survivin after the three different sectional treatments.Methods:1. The hepatoma H22 cells were subcutaneous injected to the KM mouse. The mice were divided into four groups: untreated, partial resection, PEI, and Argon-Helium cryotherapy. The survival time were analyzed by the method of Kaplean-Meier.2. The tissues from the edge of the residual tumors after three kinds of therapies were stained by the antibodies of p27, bcl-2 and survivin with the immunohistochemistrical method. The morphological changes of the cells were observed. The positive units (PU) were examined by image analysis system, and then were analyzed by the method of One-Way ANOVA and LSD.3. The tissues from both the edges of the recurrent and residual tumors after three kinds of therapies were stained by the antibodies of p27, bcl-2 and survivin with the immunohistochemistrical method. The positive units (PU) were examined by image analysis system. The differences between the positive units of the recurrent and residual tumors were compared by the method of one sample t test.Results:1. The treated mice survived for an apparently longer period after local treatments than the untreated ones. The survival time of different groups arranged from the longest to the shortest were the Argon-Helium cryotherapy group lived for 43±1 days, the PEI group for 40±l days, and the partial resection group for 36±1 days while the group without any treatment lived for 25±1 days. The results showed a significant difference among the survival time of four groups after analyzed by the method of Kaplean-Meier.2. Coagulation necrosis and cell apoptosis, shown as pyknosis, karyorrhexis and karyolysis, could be observed in some parts of the tissues after treated. The coagulation necrosis and tissue vacuole could be easily found in the sections of the Argon-Helium cryotherapy group, and a few cancer cells were alive in the necrotic area. The p27, bcl-2 and survivin proteins were positive stained as yellowish brown corpuscles. p27 was mainly found in nucleolus, while bcl-2 and survivin in cytoplasm. In the results analyzed by One-Way ANOVA, the expressions of the proteins in eachgroup were significantly different (p<0.05). The expression of p27 in cryotherapy group was higher than those in partial resection or PEI group. The expressions of bcl-2 and survivin proteins in cryotherapy group were lower than those in partial resection group. The expression of survivin protein in PEI group is lower than that in partial resection group.3. Significant difference between the recurrent and primary tumors stained by p27, bcl-2 and survivin proteins can be found in partial resection group and ethanol group (p<0.05). Among them, the expression of p27 in the recurrent tissue was lower than the primary tumor, while the expressions of bcl-2 and survivin proteins of recurrent tumors were higher. In the cryotherapy group, the expression of bcl-2 and survivin proteins of the recurrent were higher than the primary tumors (p<0.05). No significant difference of p27 was found in the cryotherapy group (p=0.056).Conclusions:1. These results showed that the left tumors in treated mice were much fewer than those in the untreated mice. Although residual tumors were still growing, the survival time of the treated mice was much longer than that of the untreated ones. Tumors in the untreated mice were larger than the treated groups at the same time and these untreated mice died more quickly. That is to say, no matter which kind of treatment is, it is very helpful to the survival time of the tumor-transplanted mice. The ones treated by Argon-Helium cryotherapy lived the longest; then comes the PEI group, and the partial group was the shortest of the three.2. After therapy, the expression of p27 of the Argon-Helium group was higher than the other two groups, while the expressions of bcl-2 and survivin were lower. That means the proliferation of the tumors after Argon-Helium cryotherapy decreased while the apoptosis increased compared with the other two groups. We presume that the malignant phenotype of the residual tumor after Argon-Helium cryotherapy would meliorate compared with the PEI or partial resection ones.3. After partial therapy, the residual tumors still grew. It was shown that the expression of P27 decreased while bcl-2 and survivin increased, which means in the recurrent tumors proliferation increased and apoptosis decreased compared with the primary ones. The cytobiological characteristics in the recurrence may have changed,and the recurrent tumors may grow faster.4. The defect of the trials is that we should be more cautious when the control groups were designed in the second and the third phase. If we had taken the samples at the same time, the results would be more stringent.5. The next study can take more indexes such as MMP-2, ICAM-1 or VEGF which can reflect more cytobiological characteristics. And the more quantized detecting methods should be used, so that the results can show more significations.
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