CAJ | 학술논문

[目的]通过观察冬凌草甲素对胰腺癌BxPC-3细胞形态和DNA变化的影响,研究其对Cyclin D/Rb/p16信号通路中基因表达的调节作用,分析冬凌草甲素对胰腺癌BxPC-3细胞的诱导与Cyclin D/Rb/p16信号通路之间的内在联系,为冬凌草甲素抗肿瘤提供可能的途径,为临床使用提供有利数据.[方法]冬凌草甲素作用BxPC-3细胞后,经瑞氏染色和Hoechst 33258荧光染色观察细胞形态,RT-PCR和实时荧光定量法(real time PCR)检测Cyclin D/Rb/p16信号通路基因的表达变化;甲基化专一性PCR试剂盒检测p16基因的甲基化.[结果]冬凌草甲素(32μg·mL-1)作用于BxPC-3细胞后,瑞氏染色可见细胞出现典型的凋亡小体,Hoechst33258荧光染色可见BxPC-3细胞出现特征性凋亡变化,冬凌草甲素(32μg·mL-1)作用BxPC-3细胞36 h时,CDK4的表达量达到最低,但p16的表达量达到最高;p16基因有甲基化.[结论]1、冬凌草甲素能诱导BxPC-3细胞形态改变.2、冬凌草甲素在一定程度下调了CDK4基因的表达,上调了p16基因的表达,不能改变p16基因的甲基化,可能通过影响Cyclin D/Rb/p16信号通路而抑制胰腺癌的生长周期.
[목적]통과관찰동릉초갑소대이선암BxPC-3세포형태화DNA변화적영향,연구기대Cyclin D/Rb/p16신호통로중기인표체적조절작용,분석동릉초갑소대이선암BxPC-3세포적유도여Cyclin D/Rb/p16신호통로지간적내재련계,위동릉초갑소항종류제공가능적도경,위림상사용제공유리수거.[방법]동릉초갑소작용BxPC-3세포후,경서씨염색화Hoechst 33258형광염색관찰세포형태,RT-PCR화실시형광정량법(real time PCR)검측Cyclin D/Rb/p16신호통로기인적표체변화;갑기화전일성PCR시제합검측p16기인적갑기화.[결과]동릉초갑소(32μg·mL-1)작용우BxPC-3세포후,서씨염색가견세포출현전형적조망소체,Hoechst33258형광염색가견BxPC-3세포출현특정성조망변화,동릉초갑소(32μg·mL-1)작용BxPC-3세포36 h시,CDK4적표체량체도최저,단p16적표체량체도최고;p16기인유갑기화.[결론]1、동릉초갑소능유도BxPC-3세포형태개변.2、동릉초갑소재일정정도하조료CDK4기인적표체,상조료p16기인적표체,불능개변p16기인적갑기화,가능통과영향Cyclin D/Rb/p16신호통로이억제이선암적생장주기.
Objective]To observe variety of morphological features, DNA band and expressions of Cyclin D/RB/p16 genes in BxPC-3 cel s treated with oridonin, investigate relationship between oridonin-induced apoptosis and Cyclin D/Rb/p16 signal pathway, which provided oridonin not only possible anti-tumor accesses but also favourable clinical data. [Methods]BxPC-3 cel s morphology was observed by Wright's staining, apoptosis was assayed by Hoechst 33258 fluorescence staining after treated with Oridonin in vitro, Cyclin D/Rb/P16 signal genes expression of BxPC-3 cel s were detected using real time PCR and a PCR kit specific of methylation for p16 gene methylation.[Results]Wright's staining showed characteristic apoptotic body in BxPC-3 cel s, Hoechst 33258 fluorescence staining showed characteristic change of apoptosis, the expression of CDK4 reduced to the minimum, and p16 gene reached the maximum; p16 gene presented methylation after treated with oridonin for 36h in BxPC-3 cel s. [Conclusion]1. Oridonin could induce BxPC-3 cel s into apoptosis. 2. Oridonin down-regulated CDK4, and up-regulated p16 and the methylation of p16 gene in BxPC-3 cel , but without effect on p16 gene methylation, which suggested that oridonin inhibited pancreatic cancer through Cyclin D/Rb/p16 signal pathway to a certain extent.