摘要
该研究的目的在于研究不同浓度华蟾素对人胃癌MKN-45细胞增殖及细胞周期的影响,分析其可能的分子机制。通过锥虫蓝拒染法检测华蟾素对细胞增殖的影响,激光共聚焦显微镜下观察细胞形态,流式细胞术检测细胞周期, qRT-PCR和Western blot分别检测细胞周期调控因子CCND1和CCNB1基因mRNA和蛋白表达情况。结果显示,在一定浓度范围内,华蟾素对人胃癌MKN-45细胞增殖的抑制作用呈时间和剂量依赖性。华蟾素作用人胃癌MKN-45细胞24 h、48 h和72 h的IC50值分别为102.797μg/L、68.939μg/L、60.563μg/L。不同浓度的华蟾素作用人胃癌MKN-45细胞48 h后,显微镜下MKN-45细胞出现明显的凋亡形态, G1期细胞所占比例降低, S期细胞所占比例逐渐升高,细胞被阻滞在S期。细胞周期调控因子CCNB1和CCND1 mRNA和蛋白表达水平显著降低。综上所述,华蟾素可以诱导细胞凋亡,阻滞细胞周期并抑制MKN-45细胞增殖。
The study investigated the effects of cinobufotalin on the proliferation of human gastric cancer MKN-45 cell.The trypan blue exclusion was adopted to detect the effect of cinobufotalin on cell proliferation,the changes of cell morphology was observed under laser scanning confocal microscopy,the change of cell cycle were detected by FACS,and the expression of CCNB1,CCND1 gene mRNA and protein were tested by qRT-PCR and Western blot,respectively.The result showed that in a certain range,cinobufotalin inhibited the proliferation of human gastric cancer MKN-45 cell,which was dependent on time and dosage.After 48 h treatment with different concentrations of cinobufotalin on human gastric cancer MKN-45 cells,the cells showed obvious apoptosis characteristics,the decrease of G1 phase and the increase of S phase cells.Compared with the control group,the mRNA and protein expression of CCND1,CCNB1 significantly decreased(P<0.05).Together all,cinobufotalin can induce apoptosis,arrest cell cycle and inhibit cell proliferation of MKN-45 cells.
作者
何孟奇
邵淑丽
冯元
焦凯贺
张伟伟
张珍珠
崔婷婷
李铁
He Mengqi;Shao Shuli;Feng Yuan;Jiao Kaihe;Zhang Weiwei;Zhang Zhenzhu;Cui Tingting;Li Tie(College of Life Science and Agriculture and Forestry,Qiqihar University,Qiqihaer161000,China;Heilongjiang Provincial KeyLaboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihaer161000,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2019年第4期674-680,共7页
Chinese Journal of Cell Biology
基金
黑龙江省自然科学基金(批准号:C201241)
黑龙江省教育厅基本业务专项重点项目(批准号:135109104)
黑龙江省教育厅基本业务专项(粮头食尾)(批准号:LTSW201737)资助的课题
黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(批准号:YSTSXK201809)~~
