In conclusion, we revealed significant telomere shortening in HCC cells and CAFs compared with their normal counterparts. More importantly, telomere shortening in cancer cells or CAFs was identified as an independent prognostic indicator for reduced survival and increased recurrence in HCC patients, highlighting the critical role of telomere dysfunction in HCC progression. As such, telomere variation in tumor cells and CAFs within the tumor microenvironment of HCC should be considered as a valuable biomarker in future clinical practice.