CAJ | 학술논문

目的:探讨紫草素是否通过诱导自噬来抑制胰腺癌细胞增殖.方法:采用噻唑蓝比色法(MTT法)研究紫草素对胰腺癌细胞Bxpc-3的增殖抑制作用.利用透射电镜观察紫草素作用胰腺癌细胞Bxpc-3后的细胞内具有双层膜结构的自噬体结构.利用荧光定量PCR(Real-time PCR)(荧光定量聚合酶链反应)和Western-blot法检测紫草素作用后胰腺癌Bxpc-3细胞自噬标记蛋白Beclin-1和LC3B的表达变化.结果:紫草素对胰腺癌细胞Bxpc-3具有明显的生长抑制作用,6μg/mL紫草素作用Bxpc-3细胞24h,其抑制率可达50%左右,且具有时间剂量依赖效应.透射电镜结果可见紫草素诱导后细胞内可见典型的双层膜结构的自噬体.Real-time PCR结果显示紫草素诱导后Bxpc-3细胞自噬相关标志物Beclin-1和LC3B表达增高.Western-blot结果显示与对照组相比,诱导后Beclin-1表达显著增高,LC3-Ⅰ表达变化不明显,但是LC3-Ⅱ/LC3-Ⅰ表达明显增高.结论:紫草素通过上调Beclin-1和LC3-Ⅱ/ LC3-Ⅰ的表达来增加胰腺癌细胞Bxpc-3的自噬活性,从而抑制胰腺癌细胞的增殖作用.
목적:탐토자초소시부통과유도자서래억제이선암세포증식.방법:채용새서람비색법(MTT법)연구자초소대이선암세포Bxpc-3적증식억제작용.이용투사전경관찰자초소작용이선암세포Bxpc-3후적세포내구유쌍층막결구적자서체결구.이용형광정량PCR(Real-time PCR)(형광정량취합매련반응)화Western-blot법검측자초소작용후이선암Bxpc-3세포자서표기단백Beclin-1화LC3B적표체변화.결과:자초소대이선암세포Bxpc-3구유명현적생장억제작용,6μg/mL자초소작용Bxpc-3세포24h,기억제솔가체50%좌우,차구유시간제량의뢰효응.투사전경결과가견자초소유도후세포내가견전형적쌍층막결구적자서체.Real-time PCR결과현시자초소유도후Bxpc-3세포자서상관표지물Beclin-1화LC3B표체증고.Western-blot결과현시여대조조상비,유도후Beclin-1표체현저증고,LC3-Ⅰ표체변화불명현,단시LC3-Ⅱ/LC3-Ⅰ표체명현증고.결론:자초소통과상조Beclin-1화LC3-Ⅱ/ LC3-Ⅰ적표체래증가이선암세포Bxpc-3적자서활성,종이억제이선암세포적증식작용.
Objective: To investigate whether Shikonin inhibits the proliferation of pancreatic cancer cells through the autophagy pathway. Methods: After Shikonin effect,the proliferation of pancreatic cancer cell Bxpc-3 was mea?sured by the MTT method, the morphologic changes of autophagy was observed with transmission electron micro?scope.After treatment with Shikonin for 24h,the expression of autophagy related genes,such as Beclin-1 and LC3B, both in mRNA and protein level,were measured by using Real-time PCR and Western-blot methods. Results: Shi?konin significantly inhibited the proliferation of Bxpc-3 cells in a dose and time dependent manner. When Bx?pc-3 cells were treated with 6μg/ml Shikonin for 24h, the inhibition rate amounted to about 50%; the typical double membrane structure of autophagosome was found under transmission electron microscope; the expression of Beclin-1 and LC3B were up-regulated compared with negative control group. Western-blot analysis further showed that the Beclin-1 and LC3B(LC3-Ⅱ/LC3-Ⅰratio) were also significantly up-regulated after treated with Shikonin for 24h. Conclusions: Shikonin can inhibit the proliferation of pancreatic cancer cells with an autopha?gy activity in pancreatic cancer cell Bxpc-3 by increasing the expression of Beclin-1 and LC3-II/LC3-I ratio.