HSPA5-associated transcriptional programs and cellular phenotypes in follicular thyroid carcinoma

Background Follicular thyroid carcinoma (FTC) is difficult to distinguish from benign follicular lesions before surgery. This study examined heat shock protein family A member 5 (HSPA5), an endoplasmic reticulum chaperone, and its association with ferroptosis-related transcriptional programs and cellular phenotypes in FTC. Methods Five public transcriptomic cohorts were integrated for candidate discovery, external characterization, and paired-tissue evaluation. HSPA5 was silenced with two short-hairpin RNA constructs in FTC-238 and FTC-133 cells. Knockdown, metabolic activity, DNA synthesis, wound closure, migration, and invasion were evaluated using molecular and cell-based assays. Results In exploratory analyses, HSPA5 correlated with ferroptosis-driver and lipid-peroxidation scores (Spearman rho = 0.709 and 0.791; false-discovery rates = 0.023 and 0.013). Expression was higher in eight patient-matched primary FTC samples than in adjacent thyroid ( P = 0.0078). External discrimination between FTC and follicular adenoma was limited (area under the receiver-operating-characteristic curve = 0.51). In both FTC-238 and FTC-133 cells, HSPA5 silencing reduced the 24-h Cell Counting Kit-8 (CCK-8) signal and the numbers of migrating and invading cells, but increased the 5-ethynyl-2'-deoxyuridine (EdU)-positive fraction. Conclusion HSPA5 is associated with ferroptosis-related transcriptional programs, and its depletion reduces 24-h CCK-8 signals and Transwell cell counts in FTC models. The role of HSPA5 in ferroptotic cell death requires functional validation.