Study On The Mitochondria Damage And Cell Apoptosis Of Hepatocarcinoma Transplanted In Nude Mice In Vivo Induced By Cadmium Chloride

Cadmium chloride is a very important heavy metal which has environmental pollution. It has a significant anti-cancer effect with a promising clinic therapy perspective, but the mechanism is still unclear. In this study, the hepatocarcinoma transplanted nude mouse model were established in vivo to study on the inhibition of the tumor by Cadmium chloride. This study can provide theoretical basis for looking for the target site of apoptosis induced by Cadmium chloride and developing low side-effect, efficient medicine.1. The anti-cancer effect of Cadmium chlorideThe hepatocarcinoma transplanted nude mouse model, were established in vivo. The average diameter of the hepatocarcinoma can achieve 1.0 cm 21 days after being transplanted. The mice were divided into 5 groups randomly. The mice in negative control group were administered with sodium chloride, the mice in positive control group were administered with 5-Fu 25 mg·kg-1, and mice in experiment groups were administered with CdC12 respectively 0.5,1.0,2.0 mg·kg-1 3 days continuously. According to the observation, the body weight of the nude mice in 1.0 and 2.0 mg·kg-1 dosage were decreased slightly in the 1st and 2nd day after administration. Then the body weight of the nude mice in 1.0 and 2.0 mg·kg-1 dosage were increased day by day; the body weight of the nude mice in other groups were increased gradually. The rate of anti-tumor tended to increase along with the elevating of CdC12 dose, and they increased significantly compared with the positive group (P<0.05),but they didn't increase significantly compared with the positive group (P>0.05).Observed under the optical microscope, the pathological changes of the hepatocarcinoma transplanted in the nude mouse were obvious with the increase of administration dosage. The hepatocarcinoma in negative group proliferated actively, just few cell thanatosis on the edge; there were many cell death in the positive group; the pathological changes of the hepatocarcinoma in mice were more and more obviously in different groups. The pathological slices were observed under the optical microscope, there were few effects on the liver, kidney, heart, lung, spleen and testis. No obvious pathological changes were observed in each slice.The mice were sacrificed by decapitated on the 10th day after administration, and took out the liver, kidney, lung, heart, spleen and testis quickly and entirely, then calculated the organ index (the organ index=the weight of each organ/the weight of body). The results revealed that compared with the PC and NC groups, The organ indexes of all experiment groups didn't vary greatly, but the kidney organ index changed significantly compared with the positive group (P<0.05).2. The biochemical changes of blood in mice administrated with Cadmium chlorideThe blood biochemical analyzer was used to analyze the blood biochemical indexes. The results were that the ALT,AST,GGT,TBIL,DBIL,A/G,UA and GLU were greatly changed in the PC group and each group administered with cadmium chloride compared with the NC group (P<0.05). These results revealed that cadmium chloride had little effect on other organs except the hepatocarcinoma, and there just were slightly biochemical changes.3. Effects of cadmium chloride on apoptosis of hepatocarcinomaTUNEL kits were used to detect the on apoptosis of hepatocarcinoma induced by cadmium chloride. Compared with the NC group, the apoptosis rate of the experiment groups increased greatly; in the 1.0 and 2.0 mg·kg-1 dosage groups the apoptosis rate changed significantly compared with the PC group. The rate of cell apoptosis tended to increase along with the elevating of CdC12 dose.4. Effects of cadmium chloride on the activity of LDH and ATPase of mitochondriaLDH exists widely in each organ of the body, and it is an important enzyme took part in the metabolism of energy. The results of this study revealed, compared with NC group, the activity of LDH decreased significantly (P<0.05) in experiment groups. The ATPase exist in the membrane of the cell and mitochondria, it can disintegrate ATP into ADP and Pi. It took important part in the information transfer, materials transportation and energy conversion. The activity of ATPase in experiment groups were higher than the negative group, and in the 1.0 and 2.0 mg·kg-1 dosage groups the activity of Ca2+-Mg2+-ATPase increased significantly compared with the negative group(P<0.05), the activity of Na+-K+-ATPase varied significantly compared with the positive group(P<0.05).5. Changes in expression of Bax, Bcl-2, Caspase-9, Caspase-3, Cyt-C of hepatocarcinomaThe immunohistochemical method was performed to detected the expression of apoptosis related proteins of the hepatocarcinoma. The expression of Bax,Bcl-2,Cyt-C,Caspase-9,Caspase-3 proteins were all increased with the dose of Cadmium chloride, and the Bax/Bcl-2 also increased gradually. Compared with the NC group, the expression of these proteins changed significantly(P>0.05).