Objective: To investigate the effects of thermal induction on transcriptional activity and targeting specificity of human telomerase reverse transcriptase promoter (hTERTp) modified with eight tandem repeats of consensus heat shock element (8HSE). Methods: The hTERTp was cloned from the genome of colon cancer cells by PCR amplification, and the hTERTp or hTERTp modified with 8HSE fragments were recombined into the dual luciferase reporter vector pGL4.2, which were then co-transfected with the internal control plasmid pGL4.74 (containing TK promoter) into the high hTERT expressing SW480 cells and low hTERT expressing MKN28 cells, respectively. The transcriptional activity of hTERTp in the two types of cells at the temperature of 37 ℃ and 43 ℃ were analyzed by dual-luciferase reporter assay system. Results: As identified by PCR, enzyme digestion and sequencing, the hTERTp cloning, 8HSE synthesis and plasmid vector constructions were all successful. Under the condition of 37 ℃, the transcriptional activity of the unmodified hTERTp in SW480 cells was significantly higher than that in MKN28 cells (P<0.05), while 8HSE modification exerted no enhancing effect on transcriptional activity of hTERTp, and it even inhibited the transcriptional activity of hTERTp in SW480 cells; under the condition of 43 ℃, the transcriptional activity of unmodified hTERTp in either type of cells showed no significant change (P>0.05), while compared with unmodified hTERTp, the transcriptional activity of 8HSE modified hTERTp in SW480 cells was significantly increased (P<0.05), but showed no significant change in MKN28 cells (P>0.05). Conclusion: Thermal induction has no obvious effect on transcriptional activity of hTERTp, but can significantly enhance the transcriptional activity of 8HSE modified hTERTp, which also specifically targets tumor cells with high hTERT expression.