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Background: Gastric cancer (GC) is among the common malignancies in the world. Studies of the KPNAs proteins’ participation in gastric cancer are still lacking. Materials and methods: The transcript levels and differential expression relating to tumor stage of KPNAs were analyzed through oncomine dataset and the Gene Expression Profile Interaction Analysis dataset. Experimental validation of mRNA and protein of KPNAs in non-malignant gastric mucosal tissues versus gastric cancer tissues was carried out using RT-qPCR, WB, and IHC. The prognostic value of KPNAs was analyzed using Kaplan-Meier Plotter. The association of KPNAs with immune invasion was assessed via the TIMER dataset. Mutations in the KPNAs family of genes were analyzed using the cBioportal dataset. In addition, inter-protein interactions expressed by the KPNAs family factors were analyzed using the String dataset, GeneMANIA. Finally, R software was adopted to analyze GO/KEGG functional enrichment of the KPNAs family factors. Results: The study illustrated that the transcript levels of KPNAs factors were elevated in GC tissues, both in mRNA and protein expression. The increased expression level of KPNA1/4/5/7 was linked to poor OS, FP and PPS. Further, it was noted that all KPNAs were linked to invasion of B-cell and the invasion of CD8+ T-cell was linked to KPNA2/3/4 expression. GO/KEGG enrichment assessment data of the KPNAs family estimated the functional role of the target host genes and biological processes showed that the observed changes were mainly cellular components, and molecular functions. Further, the cBioPortal online tool indicated that overexpression of mRNA was the most frequent alteration. Lastly, the String dataset, GeneMANIA dataset revealed that the KPNAs family factors have interactions in nuclear protein import. Conclusion: The results of the present research work showed that KPNAs might provide novel insights into potential biomarkers of prognosis along with immune invasion in GC.

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