The Effect Of Creatine Phosphate Sodium On The Growth In Mice S₁₈₀ Sarcoma And Its Mechanism

Objective: Creatine Phosphate Sodium(CP) is a medicine used to protect myocardium in many kind of heart disease. CP mainly reside in the organs needs much energy like cardiac muscle,skeletal muscle and brain.It supples the energy when cardiac cell received ischemia or hypoxia and protected cell membrane in order to avoid the injury from oxygen free radical. But because the tumor also at a high metabolism level,so there were a lot of debate on whether CP use to the person of cancer to protect the myocardium injure because of the Chemotherapy. To study the growth and apoptosis influence of CP, this experiment establish an S180 Tumor-bearing mice model and therapies with sodium chloride, CP 4㎎/d,CP8㎎/d and CP 16㎎/d ,to observe tumor weight and the tumor necrosis rate of each group were calculated,detect cell apoptosis and cell cycle and the expression of PCNA protein,Bcl-2 and Bax in the S180 transplant tumor tissue.The study is to reveal the influence of CP to the S180 transplant tumor tissue as well as to explore it's mechanism.Methods: 1.Establish an S180 Tumor-bearing mice model and treatment:60 healthy male KM mices were randomized into 4 groups : control group (sodium chloride), CP 4㎎/d group,CP 8㎎/d group and CP 16㎎/d group.Inoculate S180 cells to each mouse by right armpit subcutaneous injection.2.treatment:After the inoculation, the four groups were injected the drugs for 7 days from the next day of inoculated,once a day.All mouse were slaughted 24 hours later,subcutaneous tumors were processed for tumor weight and tumor necrosis rate.3.Use the FCM to analysis cell apoptosis,cell cycle and the expression of PCNA protein.4.The expression levels of Bcl-2mRNA and BaxmRNA were detected with semiquantity (RT-PCR).5.The expression of Bcl-2 and Bax protein in tumor tissue was qualitationly studied by immunohistochemistry method.6.Statistic analysis:All dates were showed by ( x±s) and analysis with the spss 13.0 software,including one-way ANOVA analysis and least significant difference(LSD) analysis.Results:1.Mouse solid tumer model was established successfully.2.The average weight of the tumors and inhibitory rates of the tumor: The average weight of the tumors in Control group,CP 4㎎/d group,CP8㎎/d group and CP 16㎎/d group are (2.51±0.35)g, (2.42±0.31)g,(2.35±0.28)g and (2.22±0.30)g,The weight of tumor on CP 16㎎/d group was smaller than the control group, the difference of tumor's weight between the two groups were significant (P<0.05).The difference of tumor's weight between each other groups is not significant(P>0. 05). The tumor inhibitory rate of the CP 16㎎/d group(11.60%) is the highest on the four groups, the CP 8㎎/d group(6.38%) is the second high and the CP 4㎎/d(4.25%) is the lowest。3.The result of the FCM: (1).In the treatment groups CP arrestted the cells in"G0/G1"phase, compere with the contral group (32.36±1.12%) the number of cells in"G0/G1"phase in the CP16㎎ /d group(33.30±1.34%) was ascensus, the difference is markedly(P<0. 05).The change between each other groups was not significant(P>0. 05). Cells in"S"phase were descent in the CP16㎎/d group , compere with the contral group(25.25±1.59%) there was statistically significant difference (P<0. 05) . The change between each other groups was not significant(P>0. 05). The cell apoptotic rate in CP16㎎/d group,CP8㎎/d group,CP4㎎/d group and contral group were 13.21±1.15%, 12.98±1.04%, 12.70±0.79% and 12.53±0.96%, but the difference between each group was not markedly(P>0.05). (2).The expression of PCNA protein at the saline control group(319.42±6.49) was higher than the CP 16㎎/d group(313.39±7.04),the number was statistically significant difference (P<0.05). But the difference between each group was not markedly(P>0.05). 4.RT-PCR: RT-PCR showed that with the increase of the drug,the expression of Bcl-2 mRNA and Bax mRNA in the four groups had nochange, the change between each group was not significant(P>0. 05). the ratio of Bcl-2 / Bax of CP16㎎/d group,CP8㎎/d group and CP4㎎/d group were 0.75±0.14, 0.74±0.17,0.74±0.17,(P>0.05). There are no marked change between each group(P>0.05). 5.Immunohistochemistry staining: With the increase of the drug the expression of Bcl-2 and Bax protein in the four groups also had no change, the change between each group was not significant(P>0. 05).the ratio of Bcl-2 / Bax in the control group,CP16mg/d group ,CP8mg/d group and CP4mg/d group were 0.34±0.13%, 0.31±0.11%, 0.31±0.09% and 0.32±0.12%. There were no marked change between each group(P>0.05).Conclusions: 1.High dose CP can lessen the average weight of the S180 transplant tumor; middle and low dose of CP have no effect on the average weight of the S180 transplant tumor . 2.High dose CP can inhibit the growth of the S180 transplant tumor.The cells are arrested at"G0/G1"phase in the treatment groups,"S"phase cells was descent,reduse the expression of PCNA,so inhibit the growth of S180 transplant tumor.But the middle and low dose of CP don't affect the growth of the S180 transplant tumor.3.The high dose middle dose and low dosee of CP do not affect the cell apoptotic rate,the expression of Bcl-2 or Bax and the ratio of Bcl-2 / Bax ,so CP do not influence the apoptotic of the S180 transplant tumor. 4.Provide experiment evidence for the of CP to treat the anthracycline-induced cardiotoxicity.