| Background:ALK+ NSCLC is a subtype lung cancer with a defined driver mutation after EGFR.Several m ethods,including FISH,reverse transcription PCR(RT-PCR)coupled with direct sequen cing and imm unohistochemistry(IHC),are currently used for detecting ALK rearrang ements in cytology or tissue s amples.Each technique is associated with specific strengths and weaknesses for screening,and the above methods all require histological specimens.However,it is sometimes not easy to obtain tissue sam ples in clinical practice,and it is not easy to undertake dynam ic monitoring of gene mutation status.Objective: 1.To improve the diagnostic m ethods for ALK,we compared the peptide profiles of ALK+ NSCLC and ALK-NSCLC to identif y new seru m biomarkers using MALDI-TOF-MS proteomics technology.2.We explored the use of MALDI-TOF-MS for mass spectro metric analysis of serum sa mples from advanced NSCLC patients treated with crizotinib and looked for dif ferential proteins or polypeptides that could predict efficacyMethods: 1.A total of 101 serum samples were collected: 50 sam ples from ALK+ NSCLC patients and 51 samples from ALK-NSCLC patients,all of which were examined by FISH.The peptides in the supern atants from the 50 ALK+ sam ples and 51ALK-samples were isolated with weak cation exchange m agnetic beads,and peptide peaks in the m/z range of 800–10,000 Da were analysed by MALDI-TOF-MS.The 50 ALK+ samples and 51 ALK-sam ples were randomly assigned to the training group or test group based on a 3:2 proportio n.In the training group,we screened specific serum biomarkers and established a MALDI-TOF-MS classification for ALK by comparing the protein profiles of 30 ALK+ serum samples and 30 ALK-se rum samples.The test set included 20 ALK+ serum samples and 21 ALK-serum samples to verify the results..Fin ally,we checked the accuracy,sensibility and specificity of th e diagnostic model2.Serum specimens were obtained from the advanced NSCLC patients before the trea tment of crizotinib,th e effects were evalu ated by im aging 4weeks after treatm ents,then every 8 week s.PFS was the m ain study endpoint,the patients were divided into control group and disease progression group according to the curative ef fects.Peptides were ex tracted from the sam ples using m agnetic bead-based immobilized metal ion affinity,and their mass spectra were obtained using an automated MALDI-TOF-MS system.Then we found the dif ferent peptides between control group and disease progression group and established the classification model.Results:1.The MALDI-TOF-MS identified 90 significantly dif ferent peptide peaks(P<0.05)when comparing the 30 ALK+ samples and 30 ALK-sam ples in the training group.Among the 90 peaks,the intensities of 59 peaks were upregulated in the ALK+ group,whereas the other 31 were downregulated in the ALK-group.ClinProTools software was used to sele ct 5 peptide peaks(7,477 Da,7,406 Da,7,655 Da,6,053 Da,9,307 Da)to con struct the ALK classification.The sens itivity,specificity and accu racy of the cla ssification were 7 0%,100%,and 85.7 3%,respectively,after blinded validation of the test group.2.A total of 50 patients who underwent crizotinib treatment of NSCLC patients was incl uded in the statistics,they were divided into control group(25patient s,CR+PR+SD)and disease progression group(25patients,PD)according to the curative effects.And more,the specimens were randomly divided into two groups with the ratio of 3:1.The training group was used to establish the classification m odel,this group included serum sam ples from 15 control group cases and 15 progression group cases.The test group for validating the proposed model was composed of the remaining serum samples from 10 control group cases and 10 progression group cases.There were 93 dif ferent peptide peaks,ranging from m/z 800 Da to 10,000 Da,in 15 sam ples of control group and 15 sam ples of disease progression group.41 peaks were significantly higher in the control group,as compared with the disease progression group(P<0.003,AUC?0.8).The classification model was established according to 5 peptide peaks(2,660.79?3,883.91?3,891.06?4,644.26?7,776.19?9,307.12Da),which were selected by the st atistical software.Blind testing revealed that the proposed method had 80%(8/10)sensitivity,60%(6/10)specificity,and 60%(6/10)accuracy.Conclusions: 1.In this study,we found that there were significant differences in the serum levels of ALK+ and ALK-patie nts with non-small cell lung cancer,and the different peptides may be valuable biom arkers for ALK de tection.At the sam e time,this study established a diagnostic model of se rum ALK by MALDI-T OF mass spectrometry.The model has high sensitivity and specificity and has certain clin ical application prospect.2.Application of MALDI-TOF-MS technique can select a group of serum peptides,these serum peptides expression is positively related to the efficacy of crizotinib.Background:Lung cancer is one of the most common malignancies in the world.Most patients were diagno sed are incu rable advanced or locally advanced.Lung cancer is divided into two groups,non-small cell lu ng cancer and s mall cell lung cancer,according to histo logy and m olecular biology,of which non-sm all cell lung can cer accounts for about 80-85% of the total lung cancer.It is v ery important to patients prognosis of early diagnosis.People at high risk of low dos e spiral CT screening can reduce the mortality rate of 20%.Surgical resection of the tum or remains a recommended treatment for early lung cancer.However,about 70% of patients w ith non-small cell lung cancer have been found to ha ve invasive local or distant metastasis,and therefore lose the opport unity of sur gical treatment.The progress of the disease(IIIB / IV),poor progno sis,5 years survival rate was estim ated to be about 15.9%.The clinical guidelines for patients with advan ced or locally advan ced non-small cell lung cancer recommend the two drug co mbination regimen as first-line therapy.The thirdgeneration of chemotherapeutic drugs(paclitaxel,gemcitabine,vinorelbine Changchun,docetaxel)therapy combined with platinum has becom e a standard treatm ent for advanced non-small cell lung cancer.Pemetrexed is a novel multitar geted inhibitor can inhibit the m etabolism of folic acid,f olic acid series(including enzymes of the thymidylate synthase dihydrofolate re ductase,and glycinam ide ribonucleotide formyltransferase),thus inhibiting the proliferation of tumor cells.Pemetrexed is currently recommended as first-line treatm ent of advanced non-sm all cell lung cancer drug.Purpose: To com pare pemetrexed with lipos ome pacilitaxel as f irst-line therapy in advanced non–small-cell lung cancer NSCLC.Methods: Retrospective analysis was carried on with 88 NSCLC patients of 307 hospital from July 2012 to July 2014,47 cas es of pemetrexed group and 41 cases of Paclitaxel liposome group,all first-line treatment.Study endpoint are short-term effects,PFS?OS and Main adverse events.Results:1.Short term efficacy of pem etrexed and paclitaxel liposom e in the treatm ent of non-small cell lung cancer is equivalence.The total ef fective rate is 31.9% and 21.9% respectively with no statistical diffe rence(P= 0.295);Disease control rate is 72.3%and 60.9% respectively with no st atistical difference(P = 0.258);2.PFS o f Pemetrexed in the first-line treatment of non-sm all cell lung cancer is better than paclitaxel liposome.The median PFS in the two groups was 7 m onths and 4.8 months,respectively,with statistically significant difference(P = 0.020);3.The main adverse reactions of two groups were hematologic toxicity and gastrointestinal toxicity,with no statistical difference.The incidence of alopecia were 10.6% and 39%,respectively which was lower in pem etrexed group than that of paclitax el liposome group(P<0.05).Conclusion: Short term efficacy of pem etrexed compared with paclitaxel liposome platinum in the treatment of advanced non-small cell lung cancer is equivalence.But the pemetrexed group had a survival advantage in treatment.The main adverse reactions of two groups were hematologic toxicity and gastrointestinal toxicity. |
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