Screening for regulation genes of radioresistance of oral squamous cells using lentiviral shRNA library combined with next generation sequencing / 中华放射医学与防护杂志

ABSTRACT

Objective:To screen the kinase genes related to radioresistance by high-throughput method and provide a theoretical basis for clinical treatment strategy of oral squamous cell carcinoma.Methods:The tongue squamous carcinoma cell line Cal-27 was infected with lentivirus shRNA kinase library that contains 4 675 different shRNAs regulating 709 human kinase genes. The uninfected cells were removed by puromycin screening. Then, the cells were irradiated with different doses (0, 10, 15 cGy) and continued to culture for 3 d to enrich the differences among groups. Afterwards, the cell genomic DNA was extracted and the complete shRNA sequences were obtained by PCR.Different tags were labeled in shRNAs of each group. An illumina platform was used for next generation sequencing to acquire the shRNA abundance, and then the differentiated expressed genes among different groups were identified.Results:A total of 5 kinase genes (PKLR, IPMK, AURKB, ITPKB and DLG2) were screened out from irradiated cells. Knockdown of PKLR and IPMK increased radiation resistance, while knockdown of AURKB, ITPKB and DLG2 increased radiation sensitivity, and the high expression of these genes would lead to radiotherapy tolerance.Conclusions:In this study, 3 genes relative to radioresistance of oral squamous cell carcinoma were identified by using shRNA lentivirus library combined with next generation sequencing, but the underlying mechanism requires further exploration.