Effect Of XiaoTan SanJie Decoction On Associated Proteins Of PI3K/Akt/mTOR Signal Transduction Pathway In Gastric Cancer

OBJECTIVE1.To assess the anti-tumor effects and mechanism of Chinese compound Xiaotan Sanjie Decoction on human MKN-45 gastric cancer cells lines in vivo.2.To observe the effect of Xiaotan Sanjie Decoction on associated protein expression of PI3K/Akt/mTOR signaling pathway in human MKN-45 gastric cancer transplanted on nude mice by immunohistochemical methods.3.To provide the basic evidence for the Chinese medicine Tanzheng Theory material constitution through evaluating effcet of Xiaotan Sanjie Decoction on suppression mechanism of cell proliferation and differentiation of gastric cancer.METHODS1.The nude mice model of human MKN-45 gastric cancer in situ was established.Then the experimental animals were randomly divided into four groups 48h after model established:1.the physiological saline group(control group),2.the Xiaotan Sanjie group,3.5-Fu group,4.the 5-Fu + Xiaotan Sanjie group,15 of each group.The physiological saline group was administered 0.4 ml of 0.9%saline,once a day,by gavage.The Xiaotan Sanjie group was given 0.4ml(3.354 g/ml) medicine,once a day,by gavage.The medication time of the former two groups was 6 weeks.The 5-Fu group was administered 0.4ml(6mg/ml) medicine,once a week,by intraperitoneal injection with the medication time to be 3 weeks.The 5-Fu + Xiaotan Sanjie group was given Xiaotan Sanjie Decoction medicine(0.4ml,once a day,by gavage,for 6 weeks) and was given 5-Fu at the same time(0.4ml,once a week,by intraperitoneal injection,for 3 weeks).2.The values of nude mice body weight change,tumor weight,the lump damping rate and histological change were observed in each group six weeks after initial treatment. The PI3K,Akt,mTOR and PTEN proteins expression,related to PI3K/Akt/mTOR signaling pathway,influenced by Xiaotan Sanjie on the gastric cancer were determine. The data was analyzed by Statistic method and P<0.05 was considered to have a statistic difference.RESULTS1.The body weight variation of the physiological saline group,Xiaotan Sanjie group, 5-Fu group and 5-Fu + Xiaotan Sanjie group were(5.11±0.55)g,(4.50±0.74)g, (2.02±0.37)g and(2.94±0.39)g respectively.There was a significant difference among each group(P<0.05).The mean body weight of Xiaotan Sanjie group was lower compared with the contol group but was significantly higher than those of the other two groups.The Xiaotan Sanjie group has less effect on body weight.2.The mean tumor weights were(3.121±0.403)g,(1.810±0.410)g,(1.629±0.389)g, (1.292±0.344)g in the physiological saline group,the Xiaotan Sanjie group,5-Fu group and the 5-Fu + Xiaotan Sanjie group.The inhibition lump rate of the Xiaotan Sanjie group,5-Fu group,the 5-Fu + Xiaotan Sanjie group is 41.99%,47.82%and 58.58%respectively.The mean tumor weight of Xiaotan Sanjie group is lower than that of the control goup(P=0.000,P<0.01) but has no significant statistic difference compared with 5-Fu group(P=0.203,P<0.05).There has been a better efficacy in 5-Fu + Xiaotan Sanjie group than those of other two groups(P=0.01,P<0.05).3.The positive rate of PI3K protein expression in the Xiaotan Sanjie group,5-Fu group, the 5-Fu + Xiaotan Sanjie group is 53.33%,46.67%and 53.33%respectively,which are significantly lower than that of the control group(93.33%,P<0.01).There is no significant statistic difference among the three groups(P>0.05).The PI3K protein expression in stomach cancer decreased more in Xiaotan Sanjie Decoction group than those of the other two groups.4.Xiaotan Sanjie Decoction has the good stable effect in down-regulating the Akt protein expression on stomach cancer.The positive rate of Akt protein expression in the physiological saline group,the Xiaotan Sanjie group,5-Fu group,the 5-Fu + Xiaotan Sanjie group is 86.67%,53.33%,46.67%and 40%respectively.The positive rate of Akt protein expression in Xiaotan Sanjie group has no significant statistic difference compared with those in the physiological saline group,5-Fu group and the 5-Fu + Xiaotan Sanjie group(P<0.05).The positive rates of Akt protein expression in 5-Fu group,the 5-Fu + Xiaotan Sanjie group have significant statistic difference with the physiological saline group(P<0.01).And there is no significant statistic difference between 5-Fu group and the 5-Fu + Xiaotan Sanjie group(P> 0.05).5.Xiaotan Sanjie Decoction has the good stable effect in down-regulating the mTOR protein expression on stomach cancer.The positive rate in the physiological saline group,the Xiaotan Sanjie group,5-Fu group,the 5-Fu + Xiaotan Sanjie group is 86.67%,53.33%,53.33%and 40%.The Xiaotan Sanjie group has no significant statistic difference compared with the physiological saline group,5-Fu group,and the 5-Fu + Xiaotan Sanjie group(P<0.05).5-Fu group,the 5-Fu + Xiaotan Sanjie group has significant statistic difference compared with the physiological saline group(P< 0.01).And there is no significant statistic difference between 5-Fu group and the 5-Fu +Xiaotan Sanjie group(P>0.05).6.The positive rate of PTEN protein expression in the Xiaotan Sanjie group,5-Fu group,the 5-Fu + Xiaotan Sanjie group is 80%,86.67%and 86.67%respectively, which are significantly higher than that of the control group(33.33%,P<0.01).There is no significant statistic difference among the three groups(P>0.05).The PTEN protein expression in stomach cancer increased more in Xiaotan Sanjie Decoction group than other groups.CONCLUSION1.There is a good inhibition of Xiaotan Sanjie Decoction on the human gastric cancer in nude mice MKN-45 tranplanted in situ.2.Xiaotan Sanjie Decoction plays a role on the major associated proteins of PI3K/Akt/mTOR signaling pathway,which reduces the expression of PTEN protein and enhance the phosphorylation of PI3K that leading to the activity inhibition of PI3K/AKT/mTOR signaling pathway,inhibiting tumor cell proliferation, differentiation and enhancing cell killing effects of medicine on the tumor,which may be one of the mechanism of tumor proliferation inhibition caused by Xiaotan Sanjie Decoction.3.Xiaotan Sanjie Decoction plays a role on the major proteins related to PI3K/Akt/mTOR signaling pathway,correcting the cell signal transmission, eliminating the aggregation of Qi and Tan and suppressing tumor proliferation.