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目的研究在三维细胞培养条件下前列腺细胞(RWPE-1)腺泡样结构的形成及电离辐射的影响。方法采用三维培养RWPE-1细胞,Westernblot检测自噬相关基因Beclin1及p53、p21的变化,制备Beclin1RNAi表达载体并感染细胞,免疫荧光染色和共聚焦法检测电离辐射作用后细胞形态学变化。结果三维培养条件下RWPE-1细胞聚积形成腺泡样结构(Acini),Acini组成细胞出现明显极化,2和5Gy照射后,Acini结构破坏,少量细胞发生死亡;经Beclin1基因沉默后,对照组细胞仍呈聚积趋势,2和5Gy照射后,Acini结构不再形成,细胞发生广泛性死亡。结论电离辐射抑制RWPE-1细胞Acini结构形成,Beclin1基因可能在其中发挥一定作用。
Objective To study the formation of acinar-like structures of prostate cells (RWPE-1) under three-dimensional cell culture and the effects of ionizing radiation. Methods RWPE-1 cells were cultured in three dimensions. Beclin1, p53 and p21 were detected by Western blot. Beclin1 RNAi expression vector was prepared and infected. The morphological changes of cells were detected by immunofluorescence staining and confocal microscopy. Results The acini-like cells (Acini) accumulated in RWPE-1 cells under three-dimensional culture conditions. Acini cells were obviously polarized. After 2 and 5 Gy irradiation, Acini structure was destroyed and few cells died. After Beclin1 gene silencing, The cells still showed a tendency of accumulating. After 2 and 5 Gy irradiation, the Acini structure was no longer formed and the cells died extensively. Conclusion Ionizing radiation can inhibit the formation of Acini in RWPE-1 cells, and Beclin1 may play a role in this process.