1.MET Gene Abnormalityin Chinese Patients With Lung Cancer Andtreatment Efficacy With Crizotinib:ASingle Center Retrospective Study 2.betulinic Acid Induces Potent Antitumor Effects In Paclitaxel-resistant Resistant Human Lung Carcinoma Cells (H460) Via G

Part 1.MET gene abnormality in Chinese patients with Lung Cancer and treatment efficacy with Crizotinib:A Single center Retrospective Study.Objective To investigate the clinicopathologic characteristics,treatment,and prognosis oflung cancer patients with MET gene abnormality,including 66 cases with De NovoMET amplification and 25 cases with METexon14 mutation and to guide better treatment with crizotinib.Methods We retrospective collected data from 66 cases with De Novo MET amplification and 25 cases with MET exon14 mutationfrom January 2014 to August 2019 in National Cancer Center/Cancer Hospital,Chinese Academy of Medical Sciences.All MET amplification was identified with fluorescence in-situ hybridization.Real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR)andSanger sequencing was used for METexon14 mutation,followed by next-generation sequencing(NGS;hybrid-capture targeted DNA/RNA sequencing).We then analyzed the demographic characteristics,clinical stage,histological diagnosis,prognostic characteristics of such patients and treatment efficacy and safety information with crizotinib.Results A total of 91 patients with median age of 59 years old were included in the study.The male-to-female ratio was 2.5.66 patients with median age of 59 years old were amplification of the Denovo MET in the study.The male-to-female ratio was 2.3,52%were smokers.6 patients had high-level amplification,while 32 had intermediate-level amplification and 28 had low-level amplification.The predominant subtype was adenocarcinoma.30 patients were treated with crizotinib.Of these,7 patients had a partial response,12 stable disease,and 11 progressive disease.The response rate for patientswith high,intermediate-level and low-level amplification was75%,27%and 0%,respectively.METexon14 mutation was confirmedin 25 cases,these patients with median age of 67 years old were included in this study.The male-to-female ratio was 3.2.64%were smokers and 52%had family history of cancer.Under a median follow-up of 51 months,the median OS of the whole group was 18 months.The mainly pathological typeof METex14 mutationwas asarcomatoid carcinoma.11 patients were treated withcrizotinib.Among them,10 were eligible.5 patients had a partial response,5 stable disease.The total response ratewas 50%.The median OS with amplificationofthe DenovoMET and METexon14 mutation was 21 and 13 months.Univariate analysis for MET gene abnormalityshowed that age,pathologicaltype,clinical stage in diagnosis and treatment with crizotinib had significant influence on prognosis(all p<0.05).Multivariate analysis in demonstrated that pathological type(p<0.001),clinical stage(p<0.001),treatment with crizotinib(p=0.014)were associated with prognosis.Univariate analysis showed that pathological type,clinical stage in diagnosis and treatment with crizotinib had significant influence on prognosis(all p<0.05).Multivariate analysis for amplification of the DenovoMETdemonstrated that pathological type((p<0.001),clinical stage(p<0.001),treatment with crizotinib(p=0.014)were associated with prognosis.Univariate analysis for MET exon 14 mutation showed that clinical stage in diagnosis and treatment with crizotinib had significant influence on prognosis(all p<0.05).Multivariate analysis demonstrated that clinical stage(p=0.029),treatment with crizotinib(p=0.036)were associated with prognosis.Conclusion DeNovo MET amplification could be a predictor of responsiveness to treatment with crizotinib.Treatment with crizotinib is associated with improvement in survival of patients with METexon14 mutation.Part 2.Betulinic acid induces potent antitumor effects in paclitaxel-resistant resistant human lung carcinoma cells(H460)via G2/M phase cell cycle arrest,apoptosis and alteration of Bcl-2/Bax protein signalling pathway.Objective Betulinic acid is a pentacyclic compound plant obtained from the bark of white-barked birch trees and been shown to exhibit tremendous pharmacological properties.In the present study we evaluated its anticancer potential against paclitaxel-resistant lung cancer H460 cancer cell lineMethods In the present study,the anticancer potential of betulinic acidon paclitaxel-resistant lung cancer cell line(H460)was evaluated.Cell viability was evaluated by an MTT assay,and aclonogenic assay was performed to assess the effects on cancercell colony formation.DAPI staining using fluorescence microscopy and flow cytometry were employed to evaluate the effects of betulinic acid on apoptosis.The effects of betulinic acid on the cell cycle and mitochondrial membrane potential were also evaluated by flow cytometry.The effects of betulinic acid on the protein expression of B-cell lymphoma-2(Bcl-2)/Bcl-2-associated X(Bax)were evaluated by western blot analysis.Results Betulinic acid has an IC50 value of 50 ?M against paclitaxel-resistant lung cancer cells and inhibited the colony formation potential in a dose-dependent manner.The molecule exerted its anticancer activity through induction of apoptosis by regulating Bcl-2/Bax signalling pathway.Additionally,it caused cell cycle arrest of paclitaxel-resistant lung cancer H460 cancer cells at G2/M phase.Betulinic acid was also found to cause reduction in the mitochondrial membrane potential in a dose dependent manner.Conclusion Betulinic acid may useful in the management of drug-resistant lung cancers.