放疗对人涎腺腺样囊性癌细胞的miRNA-21与细胞生物学行为的影响

The effects of radiation on miRNA-21 expression and cell biology of salivary gland adenoid cystic carcinoma cells

目的:研究放疗对人涎腺腺样囊性癌细胞SACC-83和SACC-LM的影响以及与miRNA-21表达的相关性。方法:用Varian 23 EX对培养中的SACC-83和SACC-LM予从0(对照)、2、6、10 Gy照射,CCK-8检测、流式细胞仪观察放射处理后SACC-83、SACC-LM的细胞增殖和凋亡的变化;使用qRT-PCR法分析不同剂量放疗后SACC-83、SACC-LM细胞miRNA-21的表达变化。结果:SACC-LM细胞放射处理48 h后增殖能力随放疗剂量的增大而降低(F=1321.646,P=0.000),早期凋亡率(F=354.484,P=0.039)、晚期凋亡率(F=254.278,P=0.042)和异倍体比例(F=1562.991,P=0.001)随着放疗剂量的增大而升高,且miRNA-21的相对表达量随放疗剂量的增大而减小(F=177.964,P=0.011);SACC-83细胞放疗48 h后6 Gy放疗组细胞增殖能力明显低于其他各放疗组(F=1537.214,P=0.013),6 Gy剂量下早期凋亡率(F=88.579,P=0.006)、晚期凋亡率(F=1391.345,P=0.033)、异倍体比例(F=250.461,P=0.004)最高,miRNA-21的相对表达量最低(F=134.868,P=0.017)。结论:miRNA-21可能与SACC-LM、SACC-83细胞的放疗敏感性呈负相关。

Objective： To explore the effects of radiation on miRNA-21 expression and the biology of human salivary gland adenoid cystic carcinoma cell lines（ SACC-83, SACC-LM）. Methods： In vitro cultured SACC-83 and SACC-LM cells were radiated by Varian 23 EX with the dose of 0 （control）, 2, 6 and 10 Gy respectively. Cell proliferation, apoptosis and miRNA-21 expression were observed by CCK-8 assay, flow eytometry and qRT-PCR methods respectively. Results： 48 h after radiation the proliferation ability of SACC- LM cells decreased with the increasing dose of radiation （F = 1 321. 646, P = 0.000 ）, so did the miRNA-21 expression （F = 177. 964, P = 0. 011 ）. However, the early apoptosis rate（ F = 354. 484 ,P =0.039）, the later apoptosis rate（ F = 254. 278 ,P =0.042） and heteroploid ratio（ F = 1 562. 991 ,P = 0. 001 ） increased with the increasing dose of radiation. In SACC-83 cell line 48 h after 6 Gy radiation the cell proliferation（F = 1 537. 214, P = 0.013） and the miRNA=21 expression（ F = 134. 868, P = 0.017 ） were lower than that of other radiation doses. Moreover, the early apoptosis rate（ F = 88. 579, P = 0. 006）, the late apoptosis rate（ F = 1 391. 345, P = 0.033 ）, heteroploid ratio（ F = 250. 461, P = 0.004） were higher than that of other radiation doses. Conclusion ： The miRNA-21 expression in SACC-LM and SACC-83 cell lines is conversely associated with the radiation sensitivity.