| Objective:Lung cancer is characterized by uncontrolled cell growth in lung tissue that can be transferred to nearby tissue or spread beyond the lungs.The widespread use of antitumor drugs in lung cancer treatment has led cancer cells to develop resistance to drugs.Individual difference in their sensitivity to drugs,and adaptive cancer cells can survive from drug treatment and show greater resistance.Some antitumor drugs work by binding specific proteins or molecules that inhibit the development of cancer cells and ultimately make them self-destruct.Specific targets of antitumor drugs bind to a specific protein or molecule of cancer cells,so the drug can only inhibit one pathway of the cancer cell.Before drug resistance was developed,a pathway in cancer cells was continuously suppressed and the patient's condition was controlled.This is the main reason why many cancer patients take single-target therapy drugs when there is no drug resistance.When one pathway is inhibited,the cancer cells choose other pathways to promote its own growth.Eventually,antitumor drugs will lose their therapeutic efficacy completely,making it difficult to inhibit the development of cancer cells and creating drug resistance.WZB117 is an inhibitor of glucose transporter 1,a new anticancer drug.Studies have shown that WZB117 has effect on lung and breast cancer.WZB117 can reduce cells transporting glucose,thereby reducing glucose transporter protein 1 and ATP in the cell in cancer cells and glycolytic enzyme levels,these lead to cancer cell cycle arrest,senescence and necrosis.The purpose of this study is to establish the WZB117 resistant cell lines in human non-small cell lung cancer,and to explore the resistance mechanism.Method: 1.Pretesting drug resistance successfully constructs the WZB117 resistant cell lines of non-small cell lung cancer(H1299/WZB117).2.Observing the cell morphology of H1299 and H1299/WZB117 under the microscope.3.The effects of WZB117 on the proliferation and viability of H1299 and H1299/WZB117 are tested by plate cloning and CCK8 test.4.The changes of migration capacity of WZB117 on H1299 and H1299/WZB117 are detected by scratch test.5.The effects of WZB117 on the apoptosis of H1299 and H1299/WZB117 are detected by flow cytometry.6.Rna-seq sequencing is performed to RNA from H1299 and H1299/WZB117,and analysis of the sequencing results.7.According to GO analysis results,the hypothesis of drug resistance mechanism are verified by PCR.Results: 1.Developing H1299/WZB117 successfully.2.Under the microscope,the morphology of H1299/WZB117 is compared with H1299,with increased heterogeneity,extended tentacles and elongated cells.3.WZB117 significantly inhibits the clone formation ability and vitality of H1299,and the inhibition of H1299/WZB117 is not obvious.4.WZB117 significantly inhibits the migration of H1299 and has a little inhibitory effect on H1299/WZB117.5.WZB117 induces apoptosis in H1299,and the induction of H1299/WZB117 was not obvious.6.The resistance mechanism of H1299/WZB117 is related to gene splicing,up-regulation and reduction.In H1299/WZB117,there is a change in the splicing of NUMA1.SCD and FAM198 B increase,and ID3 and ICAM1 decrease.Conclusion: In this study,H1299/WZB117 is successfully constructed,and the cell phenotype experiment shows that WZB117 can significantly inhibit the proliferation,migration and apoptosis of H1299,and the effect of H1299/WZB117 is not significant.At the same time,the mechanism of drug resistance is preliminarily discussed,which provided theoretical support for solving the problem of WZB117 resistance in clinical treatment of non-small cell lung cancer. |
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