防己诺林碱对宫颈癌HeLa细胞的作用及其机制研究  

Effects and mechanisms of fangchinoline on cervical cancer Hela cells

作　　者：蔡鑫 熊蓉 范佳杨 梁婷 汤亚兰[1] 刘康[2] 刘先平[1] 岳秋菊 罗岳西[1] CAI Xin;XIONG Rong;FAN Jia-yang;LIANG Ting;TANG Ya-lan;LIU Kang;LIU Xian-ping;YUE Qiu-ju;LUO Yue-xi(Department of Obstetrics and Gynecology,The Second Clinical Medical School of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Institute of Tissue Engineering and Stem Cells,The Second Clinical Medical School of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China)

机构地区：[1]川北医学院第二临床医学院妇产科,四川南充637000 [2]川北医学院第二临床医学院组织工程与干细胞研究所,四川南充637000

出　　处：《中国临床药理学杂志》2021年第12期1531-1534,共4页The Chinese Journal of Clinical Pharmacology

基　　金：四川省科学技术厅科技计划基金资助项目(2016JY0155);四川省教育厅基金资助项目(16ZA0239);四川省医学科研青年创新基金资助项目(Q17048);南充市校科技战略合作基金资助项目(18SXHZ0425,18SXHZ0361);川北医学院科研发展计划基金资助项目(CBY16-A-YB42)。

摘　　要：目的探索防己诺林碱(FAN)对宫颈癌He La细胞的作用及分子机制。方法人宫颈癌He La细胞随机分为空白组(0μmol·L^(-1)FAN)和低、高剂量实验组(7.5,15μmol·L^(-1)FAN)。通过CCK-8法、细胞划痕和Transwell检测FAN对宫颈癌He La细胞增殖、迁移和侵袭的影响,采用流式细胞术检测FAN对细胞周期和凋亡的作用。用Western Blot法检测细胞周期蛋白D1(Cyclin D1),P27,P53,B细胞淋巴瘤/白血病-2(Bcl2),细胞外调节蛋白激酶(ERK)和磷酸化细胞外调节蛋白激酶(p-ERK)的表达。结果FAN对宫颈癌He La细胞有明显抑制作用,呈现浓度和时间依赖关系(P<0.05)。给药48 h后,空白组、低、高剂量实验组划痕愈合率分别为(48.10±5.19)%,(22.68±3.36)%,(12.31±2.79)%;侵袭细胞数量分别为(248.60±33.34),(186.60±24.58),(53.00±13.04)个。给药24 h后,空白组、低、高剂量实验组G0/G1期细胞比例分别为(68.67±2.32)%,(77.43±3.51)%,(85.90±1.31)%;细胞凋亡率分别为(4.07±0.40)%,(5.48±1.36)%,(10.25±1.88)%;Cyclin D1蛋白相对表达量分别为0.75±0.07,0.45±0.07,0.20±0.03;P27蛋白相对表达量分别为0.01±0.00,0.17±0.02,0.55±0.04;P53蛋白相对表达量分别为0.08±0.01,0.12±0.03,0.34±0.08;Bcl-2蛋白相对表达量分别为0.40±0.03,0.17±0.02,0.06±0.01;p-ERK蛋白相对表达量分别为0.61±0.03,0.34±0.03,0.15±0.04;低、高剂量实验组的上述指标与空白组比较,差异均有统计学意义(均P<0.05)。结论FAN能抑制宫颈癌He La细胞的增殖、迁移及侵袭能力,同时阻滞细胞周期并促进细胞凋亡,其作用机制可能与抑制ERK信号通路有关。Objective To explore the effect and molecular mechanism of fangchinoline(FAN)on cervical cancer He La cells.Methods The human cervical cancer He La cells were randomly assigned to blank group(0μmol·L^(-1)FAN)and experimental-L,-H group(7.5,15μmol·L^(-1)FAN).The effects of FAN on the proliferation,migration and invasion of cervical cancer He La cells were detected by CCK-8,cell scratch and Transwell assay.And the effects of FAN on cell cycle distribution and apoptosis was detected by flow cytometry.The expressions of Cyclin D1,P27,P53,B-cell lymphoma-2(Bcl2),extracellular regulated protein kinases(ERK)and phosphorylated extracellular regulatory protein kinase(p-ERK)were detected by Western Blot.Results FAN had obvious inhibition on proliferation of cervical cancer He La cells and showed a dose and time dependence(P<0.05).There were statistically significant differences between the blank group and experimental-L/-H groups in scratch healing rates[(48.10±5.19)%,(22.68±3.36)%,(12.31±2.79)]and the number of invasive cells[(248.60±33.34),(186.60±24.58),(53.00±13.04)]at 48 h after administration;in G0/G1phase cells[(68.67±2.32)%,(77.43±3.51)%,(85.90±1.31)%],the apoptotic rates[(4.07±0.40)%,(5.48±1.36)%,(10.25±1.88)%],the relative expression of Cyclin D1 protein(0.75±0.07,0.45±0.07,0.20±0.03),the relative expression of P27 protein(0.01±0.00,0.17±0.02,0.55±0.04),the relative expression of P53 protein(0.08±0.01,0.12±0.03,0.34±0.08),the relative expression of Bcl-2 protein(0.40±0.03,0.17±0.02,0.06±0.01)and the relative expression of p-ERK protein(0.61±0.03,0.34±0.03,0.15±0.04)at 24 h after administration(all P<0.05).Conclusion FAN can inhibit the proliferation,migration and invasion of cervical cancer He La cells,while block the cell cycle and promote cell apoptosis.The mechanism may be related to the inhibition of ERK signaling pathway.

关 键 词：防己诺林碱 宫颈癌 He La细胞 增殖 迁移 侵袭 

分 类 号：R28[医药卫生—中药学]