| Objective To find the anticancer effects of Abnormal Savda Munziq(ASM)in Abnormal Savda syndrome liver cancer rat models by diong detection on 5 candidate genes.Methods Based on the Uyghur medicine theory,the Abnormal Savda rat model was induced by multi-factors,such as dry-cold feeding environment,intermittent planter point stimulation,doing forced swimming test and clipping tails for three weeks.On that basis,the liver cancer rat model with Abnormal Savda syndrome was induced by using diethylnitrosamine(DEN)for20 weeks.The rat model was intervened by high,middle and low dosages(6.0,3.0 and1.5g/kg)of Abnormal Savda Munziq(ASMq).At the end,the morphological changes of the liver specimen were observed under high power microscopic to determine whether animal models were successful,after using gene chip technology to screen out of the rat liver differentially expressed candidate p53,p21,Id4,ANG and G6 PC genes,the m RNA expressions was verified by RT-qPCR and the expressions of P53 and P21 proteins were assayed by immunohistochemical staining.Results The RT-qPCR results showed that,compared with the normal control group,the expression of p53 gene in control liver cancer group was up-regulated(P<0.01).Compared with the control liver cancer group,the expression of p53 in Abnormal Savda syndrome liver cancer group was up-regulated(P<0.05).Compared with the Abnormal Savda syndrome liver cancer group the expression of p53 was down-regulated all in 3 dosage groups,the difference was statistically significant in high and middle dosages group(P<0.05);Compared with the normal control group,the expression of p21 gene was up-regulated in control liver cancer group(P < 0.01);Compared with the control liver cancer group,the expression of p21 gene was up-regulated in Abnormal Aavda syndrome liver cancer group(P<0.05);Compared with the AbnormalSavda syndrome liver cancer group the expressions of p21 gene were down-regulated all in ASM intervened dosage groups,the middle and low dosages group were dramatic decline(P<0.01).Compared with the normal control group,the expression of Id4 gene was down-regulated in control liver cancer group(P<0.01);Compared with the control liver cancer group,the expression of Id4 gene was down-regulated in Abnormal Savda syndrome liver cancer group(P<0.01);Compared with the Abnormal Savda syndrome liver cancer group,the expressions of Id4 gene were up-regulated all in ASM intervened groups,the high and middle dosage groups were obviously up-regulated(P < 0.01);Compared with the normal control group,the expression of ANG gene in control liver cancer group was down-regulated(P <0.05);Compared with the control liver cancer group,the expression of ANG gene in Abnormal Savda syndrome liver cancer group were down-regulated(P<0.05);Compared with the Abnormal Savda syndrome liver cancer group the expressions of ANG gene were up-regulated all in 3 dosages groups(P <0.05).Compared with the normal control group,the expression of G6 PC gene in control liver cancer group was down-regulated(P<0.01);Compared with the control liver cancer group,the expression of G6 PC gene in Abnormal Savda syndrome liver cancer group were down-regulated(P < 0.05);Compared with the Abnormal Savda syndrome liver cancer group the expressions of G6 PC gene were up-regulated all in 3 dosages groups,the difference in high and middle dosages group was statistically significant(P<0.05,P<0.01).The immunohistochemistry results showed that,compared with the normal control group,the expression of P53 protein in control liver cancer group was up-regulated(P<0.01).Compared with the control liver cancer group,the expression of P53 in Abnormal Savda syndrome liver cancer group was up-regulated(P<0.05).Compared with the Abnormal Savda syndrome liver cancer group the expression of P53 was down-regulated all in 3 dosage groups,the difference was statistically significant in middle and low dosages group(P<0.01);Compared with the normal control group,the expression of P21 protein in control liver cancer group was up-regulated(P<0.01).Compared with the control liver cancer group,the expression of P21 in Abnormal Savda syndrome liver cancer group were up-regulated(P<0.05).Compared with the Abnormal Savda syndrome liver cancer group the expression of P21 was down-regulated all in 3 dosage groups(P<0.05);Conclusion(1)The occurs of Abnormal Savda syndrome liver cancer related with unusual expression of p53?p21?Id4?ANG and G6 PC genes,and the Abnormal Savda Munziq have effect on gene expression of those genes in some degree.(2)In this livercancer rat model,the Abnormal Savda Munziq plays anticancer role by doing impact on gene expression of p53?p21?ANG genes,by which the ASMq change the expression level of multiple oncogene transcription factor C-myc.(3)The Warburg effect is one of the characteristics of substance metabolism in tumor,and G6 PC is the last key enzyme in gluconeogenesis,so we sure that down-regulation of G6 PC expression in Abnormal Savda syndrome liver cancer is beneficial to the bio-synthesis of intermediates required for tumor uncontrolled growth by gluconeogenesis. |
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