Natural Anti-tumor Effect Of Pregnant Mice And Newly Born Mice And The Mechanism Of Anti-tumor Effect

Objective:⑴To investigate the natural anti-tumor effect of pregnant mice and newly born mice.⑵To investigate the effect of mice embryo extractive on proliferation, differentiation, apoptosis of H22 cells.⑶To investigate a new plan of drug combination for hepatocacinoma treatment Methods:⑴Experiment of Anti-tumor effect on pregnant mice: 15 pregnant Kunming mice were used as experimental group, and 15 non-pregnant female Kunming mice were used as control group. About 1×105 H22 cells were injected into the peritoneal cavity of each mouse, and then, the tumor growth conditions and mice's survival period were observed.⑵Experiment of Anti-tumor effect on newly born mice: 10 newly born Kunming mice were used as experimental group, 10 adult female Kunming mice were used as control group. About 750 H22 cells were injected into the peritoneal cavity of each mouse. Then, the tumor growth conditions and the mice's survival period were observed.⑶Mice hepatocarcinoma(H22) cell line was cultured in vitro. H22 cell was treated with all-trans retinoic acid (ATRA), extractive (E), 5- flurouracil (5-FU). Proliferation inhibition effect was tested by MTT assay and cell number count. In order to observe morphologic change of H22 cells, Giemsa stain was used. The activities of alkaline phosphatase(ALP) andγ-glutamyltransferase(γ-GT) were determined with colorimetry. Flow cytometry was used to detect apoptosis rate. Results : ⑴In experiment of Anti-tumor effect of pregnant mice : 5 mice of pregnant group were cured absolutely,there were no tumor growth in 60 days,the another 10 mice's mean survival period of experimental group was 36.3±1.95 days, the mean survival period of control group was 20.67±2.35 days. The experimental group's survival period is longer than the control group's (P<0.001).⑵In experiment of Anti-tumor effect of newly born mice, the mean survival period of newly born mice was 34.9±1.45 days, the mean survival period of adult mice was 24.8±2.30 days ,the newly born mice's survival period was shorter than the adult mice's (P<0.001).⑶a. Morphologic change of H22 cells: In control group, nuclear size of H22 cells is large and the nuclear-cytoplasmic ratio is high. The nuclear-cytoplasmic ratio and nuclear size of H22 cells were declined after treated with 2.5%E contrast with control group and apoptosis cells were found.b. The effect of mice embryo extractive on proliferation, differentiation, and apoptosis of H22 cells: The result of MTT experiment showed that extractive (E) could effectively inhibit the proliferation of H22 cells in a dose-dependent and time-dependent manner. The activity ofγ-GT decreased(P<0.05)and the activity of ALP increased(P<0.05)in the cells treated with 2.5% E after 120 hours compared with control group ,which means that the H22 cells differentiation degree is higher than control group's . FCM analysis showed that apoptosis rate of E group was higher than control group's (P<0.01).c. The effect of proliferation inhibition, differentiation induction and apoptosis promotion is best when 5-FU and ATRA and E used together: Cell count results show that the cell number is lowest compared with other groups when 5-FU, ATRA and E used together. The cells treated with FAE(5-FU and ATRA and E used together) had lowerγ-GT activity compare with the other groups(P<0.01 ) and the ALP activity was higher than the other groups(P<0.05 or P<0.01)and apoptosis rate was highest in all groups(P<0.01).Conclusions: ⑴There is an natural antitumor(H22 cell) effect in the body of pregnant mice and newly born mice .⑵2.5%E induced significant growth inhibition , differentiation and apoptosis of H22 cells in vitro.⑶The effect of proliferation inhibition, differentiation induction and apoptosis promotion was better when 5-FU , ATRA and E were used together.