Jin Fukang Optimization Combined With Gefitinib In Reversing The Drug Resistance And Its Mechanism In Non-Small Cell Lung Cancer

Lung cancer is one of the most malignant tumors in the world.In China,the incidence and mortality of lung cancer account for the first of the malignant tumor.At present the main clinical treatment of lung cancer by means of surgery and chemotherapy for lung cancer early,due to the lack of specific symptoms,so most of the patients have been found in advanced lung cancer,there is no chance of surgery,surgery patients with postoperative complications and recovery are also facing problems.Epidermal growth factor receptor tyrosine kinase inhibitors(e.g.gefitinib)the emergence of lung cancer patients to see the hope,but unfortunately,the patients in the use of epidermal growth factor receptor tyrosine kinase inhibitors for 8～10 months,will inevitably appear the phenomenon of drug resistance.The current situation of lung cancer treatment needs a breakthrough.TCM has many advantages,such as multiple targets,side effects,high safety and uneasy resistance.Therefore,the combination of Chinese medicine and Western medicine is more and more applied to clinical practice.This paper is mainly to discuss and study the influence of Jin Fukang optimization prescription ALG-X-012 on drug-resistant H1975 cells,so as to provide evidence and basis for further development of Chinese herbal compound,and improve a better choice for patients with non-small cell lung cancer.1.Selection of drug-resistant cell lines of gefitinibSince the molecular targeting drug gefitinib is used for a period of time in patients,the body is resistant to gefitinib,making the treatment less effective.The drug resistance of gefitinib is divided into two species,one is primary drug resistance and one is acquired resistance.Primary resistance is patients with no response to the first use of EGFR-TKI therapy in improving symptoms and control of the disease and survival time did not obtain significant benefits to;acquired resistance refers to previously treated for at least 30 consecutive days of EGFR-TKI monotherapy,with obvious clinical benefit and has the EGFR gene associated with drug susceptibility mutation treatment of EGFR-TKI.Because most patients still have disease progression within 8～12 months after EGFRTKI treatment,resulting in secondary resistance.Therefore,it is necessary to study the cause of acquired resistance,so as to provide a theoretical basis for further treatment after.The main cause of acquired resistance is the mutation of the exon 790 of EGFR gene 20,that is,T790 M mutation,which accounts for the 50% of all drug resistance mechanisms.Non small cell lung cancer cell H1975 contains both T790 M and L858 R double mutations.The EGFR-TKI acquired resistance research model as a result of T790 M mutation is widely used in.It is found that compared with PC9 cells,H1975 cell lines not only have19 exons deletion,but also have 20 exons missing,the expression characteristics of mi RNA are obvious,and H1975 cells are 100 times more resistant to gefitinib than PC9 cells.So it is reasonable to use the gefitinib resistant NCI-1975 cell line as the research object.2.The antitumor effect of Jin Fukang oral solution on ALG-X-012 in vitro This article examines the proliferation effect of Jinfukang optimization ALG-X-012 combined gefitinib on non small cell lung cancer H1975 and explore its possible mechanism.By MTT assay,scratch assay,flow cytometry experiments: optimization of ALG-X-012 and effects of gefitinib monotherapy and combination of both on H1975 cell proliferation inhibition,apoptosis and cell cycle migration;detected by Western blot method of gefitinib and gold for optimization of administration and rehabilitation alone and combined drug related protein p-EGFR on EGFR-PI3k-AKT signaling pathway pAKT,p-mTOR expression,to explore the possible mechanism.The results show:optimization of ALG-X-012 and gefitinib combination,than monotherapy group can significantly inhibit the proliferation of H1975 cells,H1975 cells and retards the migration,induce apoptosis,arrest cells in G1 phase.There was no significant difference in the expression of p-EGFR,p-AKT and p-mTOR protein between gefitinib administration group and the optimized ALG-X-012 group.Compared with the blank control group,the combination group showed a significant reduction in p-EGFR,p-AKT and p-mTOR protein compared with the blank control group.It is suggested that the mechanism of its anti-tumor activity may be related to the downregulation of p-EGFR,pAKT,p-mTOR protein expression level,thus blocking the EGFR-PI3k-AKT signaling pathway.3.The antitumor effect of Jin Fukang oral solution on the optimization of ALG-X-012 in vivoConstruction of BALB/cNCI-1975 cells in nude mice lung cancer model,to study Jifukang optimization ALG-X-012 combined with Kyrgyzstan,for the effect of non small cell lung cancer H1975 tumor by packet delivery,and through HE staining,immunohistochemical analysis and Western blotting experiments to detect tumor epithelial mesenchymal transition EMT protein expression of E-cadherin,N-cadherin,vimentin the expression of EGFR-PI3k-AKT signaling pathway and multidrug resistance related protein p-EGFR,PTEN,p-Met and p-mTOR markers and to explore its possible mechanism.The results show: optimization of ALG-X-012 group combined with gefitinib group compared with model group,administered alone group and Jifukang combined with gefitinib group,the tumor volume is smaller and the deduction rate was high and the spleen index higher tumor nude nude mice,suggesting that Jifukang optimization group combined with gefitinib group to tumor nude mice immunity.From tumor tissue sections,Jifukang optimization group and gefitinib group compared to other single drug group,the volume is smaller,less tumor cell nuclear division,the opposite is also smaller.In addition to the positive drug group,Jifukang optimization group non Kyrgyzstan expression levels of imatinib group compared to other groups significantly increased E-cadherin and PTEN,N-cadherin,Vimentin,p-Met,down-regulation of pmTOR and p-EGFR expression level,suggesting that Jifukang optimized ALG-X-012 combined with gefitinib possible mechanism of imatinib inhibits tumor growth and development and suppression of EMT the activation and blocking of EGFR-PI3k-AKT signal transduction pathway.