Investigating The Expression And Significance Of Membrane Protein GPM6B In Lung Adenocarcinoma

ObjectiveGPM6B(Glycoprotein M6B)is a transmembrane glycoprotein.It can function in several systems,especially in the nervous system where it is widely distributed,and GPM6 B is involved in myelin synthesis as well as plays an important role in neuronal function.With the increasing research on GPM6 B in tumors in recent years,we will investigate the molecular mechanism of GPM6B’s role in the development of lung adenocarcinomaMethodsFirst in xiantao academic(https://old.xiantao.love/)analysis GPM6 B expression level in various cancers.Through TCGA(https://portal.gdc.cancer.gov/)and GEO(https://www.ncbi.nlm.nih.gov/geo/)database analysis GPM6 B differentially expressed in lung adenocarcinoma.Using Kaplan Meier-Plotter database(http://kmplot.com/analysis/)on GPM6 B expression level in patients with lung adenocarcinoma and explore the relationship between the patient survival.Subsequently,the TCGA data set of lung adenocarcinoma was downloaded and GSEA enrichment analysis was conducted to explore the biological function of GPM6 B in lung adenocarcinoma.The co-expression analysis of GPM6 B in lung adenocarcinoma was conducted by using the TCGA dataset,which is convenient for the subsequent exploration of the pathway that GPM6 B may be involved in regulation.Western blot was used to detect the basic expression level of GPM6 B in HBE,A549,H1975,HCC827 and Anip973 lung adenocarcinoma cell lines.Immunohistochemical analysis of GPM6 B was performed by obtaining pathological tissue sections of lung adenocarcinoma to further explore the difference in the expression of GPM6 B in cancer and adjacent tissues.A549 and H1975 cell lines were used to construct stable overexpression GPM6 B cell lines and control cell lines by lentivirus method.Western blot and quantitative real-time PCR(q RT-PCR)were used to detect the overexpression level of GPM6 B.Subsequently,the GPM6 B overexpressed cells and the control group were subjected to functional experiments.Western blot was used to detect the changes in the expression of genes related to cell proliferation and apoptosis at the protein level.ResultsExpression of GPM6 B in various cancers and its bioinformatics analysis in lung adenocarcinoma.GPM6 B expression was significantly lower in cancer tissues than in adjacent tissues(P<0.05).The expression of GPM6 B in lung adenocarcinoma was also low(P<0.01).Survival analysis showed that GPM6 B with high expression had a better prognosis(P<0.01).GPM6 B was related to a variety of genes by co-expression analysis.GPM6 B was involved in a variety of cytological functions or pathways based on TCGA database analysis.Basal expression was detected by immunohistochemistry and Western blot.IHC showed that GPM6 B expression was significantly higher in normal tissues than in cancer tissues.Subsequently,we selected HBE and A549,H1975,HCC827,Anip973 lung adenocarcinoma cell lines to detect the difference in GPM6 B expression at protein level,and found that the expression of GPM6 b in HBE was higher than that in other lung adenocarcinoma cells(P<0.05).The efficiency of GPM6 B overexpression was determined by Western blot and RT-PCR.The protein and m RNA levels of GPM6 B were significantly increased after overexpression(P<0.05).Overexpression of GPM6 B significantly inhibited the proliferation of lung adenocarcinoma cells.Overexpression of GPM6 B in A549 and H1975 cells significantly inhibited cell growth and proliferation.In CCK8 assay,the OD value of GPM6 B overexpression group was significantly lower than that of NC group(P<0.05).The colony formation rate of GPM6 B overexpression group was lower than that of NC group,suggesting that GPM6 B could significantly inhibit the growth and proliferation of A549 and H1975.Overexpression of GPM6 B promoted apoptosis of lung adenocarcinoma cells.Western blot was used to detect the changes in the apoptotic ability of A549 and H1975 cells overexpressing GPM6 B,and the results showed that the overexpression of GPM6 B group promoted the expression of Bax and cleaved-Caspase3(P<0.05),suggesting that GPM6 B was involved in the apoptosis of lung adenocarcinoma cells.ConclusionsGPM6B plays an oncogene role in a variety of cancers,and in lung adenocarcinoma GPM6 B is associated with tumor progression,and higher expression of GPM6 B has a better prognosis.In terms of basic experiments,GPM6 B expression was higher in normal tissues than in cancer tissues,and overexpression of GPM6 B could inhibit cell proliferation and apoptosis in lung adenocarcinoma.However,there is no evidence that GPM6 B has the ability to inhibit the migration and metastasis of lung adenocarcinoma.