The expression and clinical significance of microtubule associated protein 1 light chain 3 beta in gastric cancer
Objective To analyze the expression of microtubule associated protein 1 light chain 3 beta(LC3B)in gastric cancer and its correlation with patients'prognosis based on single cell RNA sequen-cing technology.Methods Three pairs of gastric cancer and paired adjacent tissue samples were selected for single-cell RNA sequencing(scRNA-seq).CellTypist classified the filtered single cell expression data into subgroups,and Wilcoxon rank sum was used to perform quasi temporal analysis on the sequencing data to explore the differential expression of LC3B in different cell clusters.The expression of LC3B in gastric cancer tissue was analyzed using the cancer genome atlas(TCGA)and immunohistochemistry(IHC)tech-niques.Univariate and multivariate Cox proportional risk regression models were used to determine inde-pendent factors affecting the prognosis of gastric cancer patients.Real-time quantitative reverse tran-scriptase-polymerase chain reaction(RT-qPCR)was used to analyze the expression level of LC3B in gastric cancer cells,cell counting kit-8(CCK-8)assay,clone formation assay,and Transwell assay were used to detect changes in the proliferation,migration,and invasion ability of gastric cancer cells after changes in LC3B expression.Student's t-test was used to compare two groups.Results ScRNA-seq analysis revealed a significant increase in the expression of LC3B in gastric cancer tissue(2.001±2.706 vs.1.010± 1.634,t=44.094,P＜0.01).The expression of LC3B in endothelial cells and monocytes were higher than that in other subgroups(3.610±3.294,3.214±3.889,F=1 039.267,P＜0.01).Through TCGA and IHC analysis,it was found that the expression of LC3B in primary gastric cancer tissue was significant-ly higher than that in normal tissue[67.983(53.874,83.827)vs.48.114(39.147,78.174),P＜0.01;60.3％vs.19.1％,x2=9.299,P＜0.01].Patients with high LC3B expression tended to have poorer overall survival[hazard ratio(HR)=4.143,95％confidence interval(CI):2.352-7.298,P＜0.05].Multivariate Cox regression analysis showed that the expression of LC3B was a risk factor related to o-verall survival(HR=4.966,95％CI:2.241-11.005,P＜0.001).CCK-8,Transwell and clone formation analysis showed that compared with the empty body group,overexpression of LC3B significantly increased the vitality,migration,invasion and proliferation ability of gastric cancer cells[(1.837±0.057)vs.(1.287± 0.091),(338.000±39.950)vs.(152.333±4.509),(287.667±44.230)vs.(148.667±27.465),(202.333±12.503)vs.(104.667±11.240),t=8.883,7.999,4.624,10.062,P＜0.05],while inhibi-tion of LC3B expression significantly decreased the vitality,migration,invasion and proliferation ability of gastric cancer cells[(0.810±0.064)vs.(1.460±0.058),(129.667±37.859)vs.(389.667±43.822),(126.000±60.531)vs.(290.000±70.682),(54.667±16.166)vs.(109.667±3.055),t=12.963,7.776,3.052,5.790,P＜0.05].Conclusion LC3B is highly expressed in gastric cancer and promotes the proliferation and metastasis of gastric cancer cells.
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