Objective: To use microarray chip technology for screening of stem cell radiation related miRNAs in laryngealsquamous cell carcinoma; study and explore the relationship of miRNAs with radiosensitivity of laryngealsquamous cells. Method: After conventional culture and amplification of the laryngeal squamous carcinoma cellline Hep-2, CD 133+ cells were screened out with combination of isolated culture of stem cell microspheres andFACS for preparation of laryngeal cancer stem cells. After radiation treatment, miRNAs of laryngeal squamouscarcinoma stem cells before and after radiation were enriched and purified. After microarray hybridizationwith mammalian miRNA and scanning of fluorescence signal, the miRNAs of laryngeal squamous carcinomastem cells before and after radiation was subject to differential screening and clustering analysis. Real-timequantitative RT-PCR was used to verify part of the differentially expressed miRNAs. Results: 70 miRNAs relatedto laryngeal cancer stem cell radiation with 2-fold difference in expression were screened out, in which 62 weredown-regulated and 8 were up-regulated. Fluorescent quantitative RT-PCR results were consistent with miRNAschip results. Conclusion: Some miRNAs may be involved in self-regulation with laryngeal squamous carcinomastem cell radiation.