Study On Aidi Injection And PAMAM/miR-144 Synergistic Inhibition Of Gastric Cancer

Gastric cancer(gastric cancer,GC)is a malignant tumor originating from gastric mucosal epithelium,which seriously threatens human life and health.Previous studies have confirmed that the expression of miR-144 in gastric cancer is significantly reduced,has the role of tumor suppressor,can be used in gene therapy.Chinese medicine Aidi injection,including ginseng,astragalus,cantharidae,Acanthopanax senticosus,clinical often used in the treatment of malignant tumors.The combination of traditional Chinese medicine and gene therapy can enhance gene expression or increase drug sensitivity.To explore whether the Aidi Injection synergistically miR-144 anti-gastric cancer,this study first observed the effect of the Aidi Injection in patients with gastric cancer.Secondly,in order to protect the miR-144 from the degradation of nuclease,a new biological delivery system PAMAM/miR-144 was constructed by using polyamide-amine dendritic polymer material.Finally,combined with in vivo and in vitro experiments,the effects of Aidi Injection on the proliferation and migration of gastric cancer cells and the growth of subcutaneous transplanted tumor of gastric cancer in nude mice were observed.This study is divided into four parts.Objective:To collect the clinical cases of advanced gastric cancer treated with Aidi injection evaluate the curative effect from two aspects:curative effect index and TCM syndrome.Method:From October 2018 to December 2020,46 patients with advanced gastric cancer treated by Yangzhou traditional Chinese Medicine Hospital were collected.Divided into the combined treatment group of 23 cases,14 males,9 females,average age 65.09 years,treatment with Aidi Injection,Oxaliplatin and Tegafur.Chemotherapy group,23 cases,12 males,11 females,average age 67.91 years,treatment with Oxaliplatin and Tegafur.Observe the curative effect from the index and TCM syndrome evaluation.Results:There was no statistically significant difference in gender,age and TNM stage of the two groups between the two groups(P>0.05).Compared with the control group,the combined group had a higher quality of life score(P<0.05),a higher effective rate of TCM syndrome scores(P<0.05).Conclusion:Aidi injection can increase the therapeutic effect of chemotherapeutic drugs on advanced gastric cancer and improve patient’s symptoms.Objective:To construct a stable PAMAM/miR-144 delivery system of encapsulated miR-144 and to detect its physicochemical properties and biological activities.Methods:Polyamide-amine-type dendritic polymer materials(PAMAM,1.25,2.5,5,10 mg/ml)were mixed proportionally with 10 nmol/ml miR-144,and the expression of miR-144 was detected by qRT-PCR to determine the optimal proportion.The shape and particle size of the PAMAM/miR-144 of the biological delivery system were detected under transmission electron microscopy.At the same time,the potential and charge of the PAMAM/miR-144 complex are detected.The expression of mammalian target protein(mTOR)in human gastric cancer AGS cells and human gastric cancer HGC-27 cells was detected by qRT-PCR.Results:The PAMM/miR-144 is a homogeneous and circular stable complex with a particle size of(254.66±5.67)nm.And PAMAM/miR-144 significantly decreased the expression of mTOR in human gastric cancer AGS cells and human gastric cancer HGC-27 cells.Conclusion:PAMAM/miR-144 complex can deliver miR-144,stably and effectively and has biological activity.Objective:To study the effect of Aidi Injection synergistic PAMAM/miR-144 on the proliferation and the migration in human gastric cancer HGC-27 cells,and to explore its molecular mechanism.Methods:MTT method was used to detect the effects of different concentrations of Aidi Injection(7.5,15,30,60,120 mg/ml)on the proliferation of human gastric cancer HGC-27 cells.The effect of Aidi Injection combined with PAMAM/miR-144 on the migration ability of human gastric cancer HGC-27 cells was observed.The protein expressions of PI3k,Akt,mTOR,and 4EBP1 were detected by using Western Blotting in human gastric cancer HGC-27 cells.Results:Aidi Injection significantly inhibited the proliferation of human gastric cancer HGC-27 cells in a time-and concentration-dependent manner.On the basis of the IC50 value,the concentration of Aidi Injection was determined to be 18 mg/mL in the subsequent experiment.The migration ability of human gastric cancer HGC-27 cells was significantly inhibited by Aidi Injection combined with PAMAM/miR-144.The protein expression levels of PI3k,Akt,mTOR,and 4EBP1 were significantly decreased.Conclusion:Aidi Injection combined with PAMAM/miR-144,significantly inhibits the proliferation and migration of human gastric cancer HGC-27 cells,and its molecular mechanism may be related to the inhibition of PI3k/Akt/mTOR signal transduction pathway.Objective:To study the effect of Eddie injection combined with PAMAM/miR-144 on the growth of subcutaneous transplanted tumor of gastric cancer in nude mice.Methods:The concentration of human gastric cancer HGC-27 cells was adjusted to 107/ml.0.2 ml cell suspension was mixed with matrix glue and inoculated under the back of nude mice.After the transplanted tumor model was established successfully,the mice were randomly divided into sterile saline control group,Aidi Injection group,PAMAM/miR-144 group,Aidi Injection combined with PAMAM/miR-144 group,10 mice in each group.Each group was given according to the requirements.Observe the general condition of spirit and diet of nude mice.At the end of the experiment,nude mice were sacrificed,the tumor was removed and the tumor volume,size and weight of each group were compared.Results:Compared with the control,Aidi Injection combined with PAMAM/miR-144 significantly inhibited the weight of tumor more significantly(P<0.05),and the volume of tumor was smaller(P<0.05)..Conclusion:Aidi Injection combined with PAMAM/miR-144 can inhibit the growth of subcutaneous transplanted tumor in nude mice.