Molecular Diagnosis Of LKB1 Gene Mutations And Cell Autophagy In Lung Cancer

Lung cancer has become a disease with high incidence rate all over the world.Every year,about 10000 people are diagnosed with lung cancer,and 80000 people die of this disease.Although rapid progress has been made in the diagnosis,classification,and treatment of lung cancer,the overall survival rate of lung cancer is still very poor,and the prognosis of patients is still unfavorable.In the past few decades,many evidences have shown the importance of liver kinase B1(LKB1)as a tumor suppressor gene in the development and progression of solid tumors.LKB1,as a powerful biomarker representing the functional status of tumors,can guide the prognosis evaluation of patients.However,the diagnostic and clinical value of LKB1 in patients with lung cancer is controversial.Therefore,this article designs and constructs a laboratory LKB1 detection system,collects 22 lung cancer patients from January 2021 to June 2022,and extracts the genomic DNA of 22 patients after signing an informed consent form.Primers are designed for the design of LKB1 full exons,and the extracted genomic DNA is amplified in full exons.The first generation sequencing method is used for sequencing.The sequencing results are compared through NCBI-BLAST to identify mutations in the base sequence,Based on this,a laboratory mutation detection system for LKB1 gene was established,and the self built LKB1 gene mutation detection system was systematically validated.Using the original data of the gene chip numbered GSE108214 in the GEO database,download the original data and conduct bioinformatics analysis to study the analysis of the drug resistance and toxicity of cisplatin on LKB1 mutant cell lines.The LKB1 mutant cells were treated with cisplatin and isoginkgo biloba biflavones,and the autophagy related pathway proteins in the LKB1 mutant cells were detected by Western Blot method.The experimental results showed that the mutation detection system for LKB1 gene constructed in this experiment sequenced the entire exon of LKB1 in 2 2 patients,and detected nonsense mutations in the non coding region of 3 patients,while 19 patients had no mutations.This system can be used to detect mutations in LKB1 gene in lung cancer patients and detect clinical samples.At the same time,there was a non-toxic analysis of LKB1 mutations treated with cisplatin,with significant upregulation of differential genes.Functional analysis of GO genes mainly included glutamate receptor signaling pathways,ribonucleotide metabolism processes,and so on.KEGG signal pathway enrichment is mainly annotated to P53 signal pathways,etc.Analysis of cisplatin resistance and sensitivity of LKB1 mutant cells treated with cisplatin revealed significant upregulation of differential genes.Functional analysis of GO genes mainly included the regulation of cell morphology,cell growth,and so on.The enrichment of KEGG signaling pathways is mainly annotated with focal adhesion.In addition,the results of cisplatin combined with isoginkgo biloba biflavones on the regulation of autophagy in LKB1 mutant cell lines showed that.Cisplatin combined with isoginkgo biloba biflavone can upregulate and protect the expression of tumor suppressor gene p53,but Caspase3 gene regulation is not obvious,inhibiting Bcl-2 upregulation of Bax,and promoting cell apoptosis.After combined treatment,the expression of tumor suppressor LKB1 was upregulated,which could inhibit m TOR expression,thereby inducing autophagy,activating the expression of p-JNK and ERK.Cisplatin 10 u M could inhibit 4EBP1 and p-4EBP1,and inhibit the synthesis of cell growth proteins.The above results indicate that this study can detect mutations in the LKB1 gene in clinical samples of lung cancer through a laboratory built system.Cisplatin combined with isoginkgo biloba biflavones can promote cell apoptosis,autophagy,and protein synthesis.This will provide treatment options and new strategies and approaches for lung cancer with LKB1 mutations in the future.