Department of Oncology, Wannan Medical College, Wuhu 241001, China.
Fitoterapia. 2011 Dec;82(8):1258-64. doi: 10.1016/j.fitote.2011.08.015. Epub 2011 Sep 1.
To investigate the anti-tumor effects of L-securinine inducing colon cancer SW480 cell autophagy and explore its potential molecular mechanism.
MTT method was used to detect the antitumor effect of SW480 cells cultured with L-securinine in vitro. Light and electron microscopy were used to observe SW480 cells treated with L-securinine morphological changes. Flow cytometry was used to observe the apoptoticratio and cell cycle inducing with the L-securinine in SW480 cells, and the autophagic apoptosis ratio was determined by FITC-conjugated annexin V by flow cytometry (FCM). FCM was applied to analysis cell cycle; the expression of autophagy gene Beclin-1 was examined by reverse transcriptase polymerase chain reaction (RT-PCR).
The generation depression effects of SW480 cells cultured in vitro were detected byMTT method (Pb0.05), and there were dosage-time dependent relationships. Numerous autophagic vacuoles and empty vacuoles were observed in SW480 cells treated with 2.5 μM L-securinine for 48 h by electron microscopy, and the process of cell division that got less was observed.Through flow cytometry, a number of observed autophagic cells were obviously increased, and G1/S phase was retarded. L-Securinine tended to arrest cells at the G1 phase of the cell cycle. The percentage of the apoptotic cells increased as treatment duration and concentrations increased. Beclin-1 expression enhanced with L-securinine concentration increased.
L-Securinine has an anti-tumor effect against colon cancer SW480 cell. The L-securinine can induce striking autophagy in SW480 cell in vitro. The autophagy induced by L-securinine is related with upregulating the expression of autophagy gene Beclin-1.
探讨 L-苦参碱诱导结肠癌 SW480 细胞自噬的抗肿瘤作用，并探讨其潜在的分子机制。
采用 MTT 法检测 L-苦参碱体外培养 SW480 细胞的抗肿瘤作用。光镜和电镜观察 L-苦参碱处理 SW480 细胞的形态变化。流式细胞术观察 L-苦参碱诱导 SW480 细胞凋亡率和细胞周期，用 FITC 标记的 annexin V 通过流式细胞术（FCM）检测自噬凋亡率。FCM 分析细胞周期；逆转录聚合酶链反应（RT-PCR）检测自噬基因 Beclin-1 的表达。
MTT 法检测到体外培养的 SW480 细胞的生成抑制作用（Pb0.05），且存在剂量-时间依赖性关系。电镜下观察到 2.5 μM L-苦参碱处理 48 h 的 SW480 细胞中出现大量自噬空泡和空泡，观察到细胞分裂过程减少。通过流式细胞术，观察到明显增加的自噬细胞数量，G1/S 期延迟。L-苦参碱倾向于使细胞周期停滞在 G1 期。随着处理时间和浓度的增加，凋亡细胞的比例增加。Beclin-1 的表达随着 L-苦参碱浓度的增加而增强。
L-苦参碱对结肠癌 SW480 细胞具有抗肿瘤作用。L-苦参碱能在体外显著诱导 SW480 细胞自噬。L-苦参碱诱导的自噬与上调自噬基因 Beclin-1 的表达有关。
