Previous studies have shown that heat shock proteins (HSPs) were upregulated in various types of tumors and were associated with histological grade, recurrence and metastasis of malignant tumors. In this study, we investigated whether heat shock protein 70 kDa (HSP70) was associated with histological grade of laryngeal squamous cell carcinomas (LSCC). We also determine the role of HSP70 in LSCC radiation resistance using a laryngeal carcinoma xenograft model by antisense HSP70 RNA technique. Immunohistochemistry data showed that HSP70 was detected in 96% of LSCC tissues (48 out of 50). The expression level of HSP70 was significantly lower in early stage of LSCC than that in late stage (P = 0.015). Radiation treatment result showed that the volumes and weights of implantation tumors in the group injected with antisense HSP70 oligos were significantly reduced comparing to the group injected with random oligos(p < 0.05). In addition, cleavage and degradation of tumor nucleolin in antisense HSP70 oligos injection group was significantly higher than that in random oligos injection group. Our result suggested that HSP70 may play a role in LSCC radiotherapy resistance by inhibiting cleavage and degradation of nucleolin.