Study On The Effect Of Yiqi Jianpi Kangcan Prescription On Gastric Cancer-bearing Nude Mice Based On JAK2/STAT3 Signaling Pathwa

Objective:The effects of Yiqi Jianpi anticancer formula and Its Anticancer Traditional Chinese medicine on Janus activated kinase 2（JAK2）,signal transducer and activator of transcription 3（STAT3）,phosphorylation signal transducer and activator of transcription 3（p-STAT3）and hypoxia inducible factor 1 in gastric cancer（GC）bearing nude mice were studiedα（hypoxia inducible factor-1α,HIF-1α）To explore the mechanism of Yiqi Jianpi anticancer formula and its anticancer drugs on GC,so as to provide experimental basis for the clinical application of Yiqi Jianpi anticancer formula and its anticancer drugs in the treatment of gastric cancer.Materials and methods:Human gastric cancer cell SGC-7901 cell suspension was subcutaneously inoculated into the right armpit of nude mice to establish the subcutaneous transplanted tumor model of nude mice.After the model was successfully established,the nude mice were divided into model control group,S-1 group,traditional Chinese medicine 1combined with S-1 group（traditional Chinese medicine 1:Yiqi Jianpi anticancer formula reduction protection formula）,traditional Chinese medicine 2 combined with S-1 group（traditional Chinese medicine 2:Yiqi Jianpi anticancer formula constant formula）;The model control group was treated with oral gavage of normal saline,0.2ml each,once a day for 15days;In S-1 group,0.1ml of S-1 was administered by gavage(10mg·kg^-1·D^-1)once a day for 15 days;According to the equivalent dose relationship converted from human and rat body surface area,traditional Chinese medicine 1 combined with S-1 group(the concentration of traditional Chinese medicine 1 is 403mg·kg^-1·D^-1)was gavaged once a day for 15days;Traditional Chinese medicine 2 combined with S-1 group(the concentration of traditional Chinese medicine 2 is 481mg·kg^-1·D^-1)was administered by gavage once a day for 15 days,and the materials were taken the next day after the last administration.Test indexes:the weight change of nude mice before and after administration and the tumor weight after stripping tumor tissue were weighed by electronic scale,and the tumor inhibition rate was calculated;Data were used to analyze the volume of transplanted tumor at 1,3,6,9,12and 15 days;Western blot（WB）and immunohistochemistry were used to detect JAK2,STAT3,p-STAT3 and HIF-1αin transplanted tumor tissues of gastric cancer in nude mice Protein expression.SPSS 26 statistical software was used to analyze the experimental data,and there was significant difference（P<0.05）.Results:1.Tumorigenesis rate:on the 10th day after inoculation,the maximum tumor diameter of nude mice is≥5mm,which is regarded as successful modeling.35 of 40 nude mice have tumorigenesis,and the tumorigenesis rate is 87.5%.2.General state of nude mice:at the later stage of gavage,the nude mice showed obvious weight loss,poor spirit,reduced activity and reduced diet,similar to cachexia.With the increase of volume,the subcutaneous tumors of individual nude mice broke and scabbed on the surface.3.Changes in body weight and tumor volume of nude mice before and after administration:after the drug intervention,nude mice in each group showed varying degrees of weight loss,decreased activity and diet,and reduced body weight compared with those before administration（P<0.05）.After gavage,the tumor volume of each group was:model control group（792.58±64.646）mm³;S-1 group（673.503±60.366）mm³;TCM 1 combined with S-1 group（652.095±56.474）mm³;Traditional Chinese medicine 2 combined with S-1 group（479.43±33.677）mm³.Compared with the model group,the tumor volume in the drug intervention group was significantly reduced（P<0.05）,4.Tumor weight and tumor inhibition rate:after treatment,the tumor weight of each group was:model group（2.048±0.225）g;S-1 group（1.815±0.185）g;Traditional Chinese medicine 1 combined with S-1 group（1.807±0.175）g;Traditional Chinese medicine 2combined with S-1 group（1.583±0.156）g;Compared with the model control group,the tumor weight of each treatment group was significantly reduced（P<0.05）.Compared with single drug S-1,the tumor weight of traditional Chinese medicine 1 combined with S-1 was slightly reduced,but there was no statistical significance（P>0.05）;Traditional Chinese medicine 2 combined with S-1 group was less than other groups（P<0.05）.The tumor inhibition rates of S-1 group,TCM 1 combined with S-1 group and TCM 2 combined with S-1 group were 11.2%,11.6%and 22.6%respectively.5.Protein expression:the results of Western blot and immunohistochemistry showed that JAK2,STAT3,p-STAT3 and HIF-1αin tumor tissues of each administration group compared with the model control group The protein expression decreased（P<0.05）;There was significant difference between S-1 group and TCM 1 combined S-1 group（P<0.05）;Compared with the other two groups,the protein expression of traditional Chinese medicine 2combined with S-1 group decreased（P<0.05）;The protein expression of TCM 1 combined with S-1 group and TCM 2 combined with S-1 group was statistically significant,which may be that the dose of Anticancer Traditional Chinese medicine affected the results.Conclusion:In the transplanted tumor model of human gastric cancer in nude mice,S-1group,traditional Chinese medicine 1 combined with S-1 group and traditional Chinese medicine 2 combined with S-1 group inhibited the growth of subcutaneous transplanted tumor in nude mice to varying degrees,that is,S-1 and Yiqi Jianpi anticancer formula played an anti-tumor role to a certain extent;Compared with S-1 alone,the effect of the group containing traditional Chinese medicine was significantly improved,indicating that traditional Chinese medicine can play a synergistic role;The effect of traditional Chinese medicine 2 is more obvious than that of traditional Chinese medicine 1,which further shows that the anti-tumor effect of Yiqi Jianpi anticancer constant formula is more obvious;The body weight of nude mice in TCM 2 plus S-1 group is higher than that in TCM 1 plus S-1 group,which shows that the combination of Yiqi Jianpi anticancer constant formula and S-1 can play a good role in expelling evil and strengthening health;The results of immunohistochemistry and Western blot showed that Yiqi Jianpi anticancer formula could down regulate JAK2,STAT3,p-STAT3 and HIF-1αThe possible mechanism of protein expression level is to down regulate JAK2,STAT3,p-STAT3 and HIF-1αby affecting JAK2/STAT3 signaling pathway The level of protein expression,so as to inhibit the proliferation,invasion and metastasis of gastric cancer cells.