Abstract: Objective :To assess the effect of RNA interference on the human telomerase reverse transcriptase (hTERT) mRNA in treatment of laryngeal squamous carcinoma. Methods :Two types of plasmids, pshRNAl and pshRNA2, as will as the involved ffuorescein gene, were synthesized accordingto the structures of hTERT cDNA. The tumor model of nude mice was established by subcutaneous implantation of Hep-2 cells. shRNA plasmids wire transfected into Hep-2 cells and the transplantation tumors. Fluorescence expression in Hep-2 cells and the tumor was detected by confocal microscopy.The proliferation of Hep- 2 cells was detected by M7 assay. Confirmation of hTERT protein expression in vitro and in vivo was performed by Western Blotting and immunohistochemistry, respectively. Results :After the Hep-2 cells and the tumors were transfected by pshRNAl and pshRNA2, many greenfluorescent cells were observed by confocal microscopy. The viability of Hep-2 cells was inhibited by pshRNAl and pshRNA2. pshRNAl and pshRNA2 could detect a low level of TERT protein expression in Hep-2 cells and the transplantation tumors. Studies of in vivo experiment demonstrated thatpshRNAl significantly inhibited the growth of the established tumors in nude mice. Conclusions :results demonstrate shRNA plasmids, which express specific sequences of hTERT gene, are effectively transfected into Hep- 2 cells in vivo and in vitro. This data can also support the potential application ofRNA interference to treat the laryngeal squamous carcinoma.