Heat-shock proteins (Hsps) are a family of proteins essential in preserving the vitality and functionality of proteins under stress conditions. Cucumber (Cucumis sativus) is a widely grown plant with high nutritional value and is used as a model organism in many studies. This study employed a genomics, transcriptomics, and metabolomics approach to investigate cucumbers' Hsps against abiotic stress conditions. Bioinformatics methods were used to identify six Hsp families in the cucumber genome and to characterize family members. Transcriptomics data from the Sequence Read Archive (SRA) database was also conducted to select CsHsp genes for further study. Real-time PCR was used to evaluate gene expression levels under different stress conditions, revealing that CssHsp-08 was a vital gene for resistance to stress conditions; including drought, salinity, cold, heat stresses, and ABA application. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of plant extracts revealed that amino acids accumulate in leaves under high temperatures and roots under drought, while sucrose accumulates in both tissues under applied most stress factors. The study provides valuable insights into the structure, organization, evolution, and expression profiles of the Hsp family and contributes to a better understanding of plant stress mechanisms. These findings have important implications for developing crops that can withstand environmental stress conditions better. •Heat Shock Protein Family members in Cucumis sativus genome were identified and characterized with bioinformatics methods.•Real-time PCR showed that the expression of CssHsp-08, CsHsp40-70, and CsHsp70-06 genes significantly increased in response to different stress conditions.•Metabolite level analysis revealed that the accumulation of amino acids in leaves and sucrose accumulation in both tissues were observed under most stress factors.•The work gives crucial knowledge on the organization, metabolite, and expression profiles of the cucumber heat shock protein family under stress conditions.