CAJ | 학술논문

金钗石斛生物碱(Dendrobine)对肿瘤细胞具有抑制作用,但其对于乳腺癌细胞还未见报道.通过MTT实验,发现乳腺癌细胞的生长活力受到金钗石斛生物碱的明显抑制.对乳腺癌细胞进行Annexin V-FITC/PI染色流式细胞术(Flow cytometry,FCM)检测细胞凋亡率,进一步说明金钗石斛生物碱对乳腺癌肿瘤细胞生长的抑制作用,且具有明显的促凋亡效应.实验结果显示金钗石斛生物碱抑制了mcf-7细胞分裂过程中从G1期向S期的转换.JC-1探针荧光共聚焦显微镜观察细胞线粒体跨膜电位变化,红绿荧光比例变化由低变高.荧光免疫印迹法(WensternBlot)测定细胞半胱氨酸天冬氨酸蛋白酶(caspase-3)凋亡基因蛋白表达量发现,procaspase-3和被剪切的cleaved caspase-3蛋白表达条带均清晰可见,说明通过线粒体途径激发了细胞凋亡执行基因半胱氨酸天冬氨酸蛋白酶,使得癌细胞发生凋亡.实验结果显示金钗石斛生物碱能使mcf-7细胞通过线粒体途径发生细胞凋亡,且有细胞色素C释放,并激活procaspase-3启动了细胞的死亡程序.
금차석곡생물감(Dendrobine)대종류세포구유억제작용,단기대우유선암세포환미견보도.통과MTT실험,발현유선암세포적생장활력수도금차석곡생물감적명현억제.대유선암세포진행Annexin V-FITC/PI염색류식세포술(Flow cytometry,FCM)검측세포조망솔,진일보설명금차석곡생물감대유선암종류세포생장적억제작용,차구유명현적촉조망효응.실험결과현시금차석곡생물감억제료mcf-7세포분렬과정중종G1기향S기적전환.JC-1탐침형광공취초현미경관찰세포선립체과막전위변화,홍록형광비례변화유저변고.형광면역인적법(WensternBlot)측정세포반광안산천동안산단백매(caspase-3)조망기인단백표체량발현,procaspase-3화피전절적cleaved caspase-3단백표체조대균청석가견,설명통과선립체도경격발료세포조망집행기인반광안산천동안산단백매,사득암세포발생조망.실험결과현시금차석곡생물감능사mcf-7세포통과선립체도경발생세포조망,차유세포색소C석방,병격활procaspase-3계동료세포적사망정서.
Dendrobine from Dendrobium nobile could wield an inhibitory effect on cancer cell,but the in-formation on its treatment of breast cancer is not available so far.By the MTT experiment,it was found that den-drobine could obviously restrain the growth of the breast cancer cells mcf-7.The apoptosis rate was determined with Annexin V-FITC/PI staining based on flow cytometry (FCM) on mcf-7.The inhibitory effect and apopto-sis-promoting effect of dendrobine on mcf-7 were uncovered further.The results indicated that dendrobine inhib-ited the transition from G1 to S in the process of the division of mcf-7 cells division.Mitochondrial transmembrane potential variation in the cells was observed with JC-1 probe and fluorescence confocal microscopy.It showed that the Red/Green fluorescence ratio of mitochondrial membrane potential in MCF-7 cells changed from low to high.The amount of protein expression of apoptotic gene caspase-3 was detected with Western Blot.The bands of pro-caspase-3 and cleaved-caspase-3 could be clearly observed.The results indicated that the cell apoptosis could be induced by apoptotic gene caspase-3 and it involved the activation of mitochondrial pathway.It is concluded that dendrobine can cause apoptosis of the cells of breast cancer,and the main pathway is through mitochondria towards apoptosis,where the cells released the cytochrome C and invoked the procaspase-3.