胃癌细胞株SGC-7901诱导凋亡过程中细胞外RNA测定

Detection of extracellular RNA in supernant of artificially-indued apoptosis of SGC-7901 cell line

目的:研究胃癌细胞株SGC-7901细胞外RNA稳定性,探讨细胞外RNA来源。方法:将胃癌细胞株SGC-7901分为正常培养组、过氧化氢(H2O2)诱导坏死组和地塞米松(DEX)诱导凋亡组,巢式PCR测定各组不同时间点细胞上清液中RNA含量;应用RNA酶等处理上述细胞上清液,观察每组释放到细胞外RNA的稳定性。结果:正常培养组细胞外RNA在30 m in即可被检测到,随后逐渐增加,8 h后维持稳定;诱导坏死组和凋亡组在4 h之后细胞外RNA的量均有明显的增加,12 h后在高水平维持稳定。细胞凋亡组细胞外RNA有很强的稳定性,正常培养组和坏死组细胞外RNA则容易降解。结论:凋亡细胞释放的RNA有着特殊的稳定性,可能是被检测到的血液中RNA的主要来源。

Objective： To observe the stability of extracellular RNA（exRNA） in supernant of culture SGC-7901 cell line,explore the origin of exRNA.Methods： The cell were divided into three groups,the normal group,H2O2-induced necrosis group and DEX-induced apoptosis group.Nested-PCR was used to detected changes of RNA quantity at different time points in supernatant of normal culture and artificially-induced necrosis and apoptosis.In order to study the stability of the exRNA,RNAse were added to the samples and incubated for 1 hour at 37 ℃ before RNA extraction and subsequent RT-PCR.Results： ExRNA was detectable at 30 min after changing the medium and reached a plateau at 8 h.Compared with the normal group,the amount of exRNA in artificially-induced necrosis and apoptosis groups had a significant increase at 4 h after changing the medium and reached a high plateau at 12 h.The exRNA had high stability in apoptosis group,and was easily degradable in normal and necrotic cell group.Conclusion： The RNA released by apoptotic cells was stable and may be the main source of exRNA in blood.