[1]杨晓瑞,王　江,潘　琼,等.奥希替尼联合人参皂苷Rg3对非小细胞肺癌H1975细胞增殖和凋亡的影响[J].新乡医学院学报,2020,37(11):1007-1012.[doi:10.7683/xxyxyxb.2020.11.002]
　YANG Xiaorui,WANG Jiang,PAN Qiong,et al.Effect of osimertinib combined with ginsenoside Rg3 on proliferation and apoptosis of non-small cell lung cancer H1975 cells[J].Journal of Xinxiang Medical University,2020,37(11):1007-1012.[doi:10.7683/xxyxyxb.2020.11.002]
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奥希替尼联合人参皂苷Rg3对非小细胞肺癌H1975细胞增殖和凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年11
页码:
1007-1012
栏目:
基础研究
出版日期:
2020-11-05

文章信息/Info

Title:
Effect of osimertinib combined with ginsenoside Rg3 on proliferation and apoptosis of non-small cell lung cancer H1975 cells
作者:
杨晓瑞1; 王　江2; 潘　琼1; 刘凡凡1; 丁孝良1
（1.郑州大学附属郑州中心医院药学部，河南　郑州　450000；2.郑州大学附属郑州中心医院胃肠疝和腹壁外科，河南　郑州　450000）
Author(s):
YANG Xiaorui1; WANG Jiang2; PAN Qiong1; LIU Fanfan1; DING Xiaoliang1
(1.Department of Pharmacy,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,Henan Province,China;2.Department of Surgery of Gastrointestinal Hernia and Abdominal Wall,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,Henan Province,China）
关键词:
奥希替尼; 人参皂苷Rg3; 非小细胞肺癌; 增殖; 凋亡
Keywords:
osimertinib; ginsenoside Rg3; non-small cell lung cancer; proliferation; apoptosis
分类号:
R734.2
DOI:
10.7683/xxyxyxb.2020.11.002
文献标志码:
A
摘要:
目的　探讨奥希替尼联合人参皂苷Rg3对非小细胞肺癌H1975细胞增殖、凋亡的影响及其机制。方法　将对数生长期的H1975细胞随机分为人参皂苷Rg3组、奥希替尼组、奥希替尼+人参皂苷Rg3组和对照组，每组设5个复孔。人参皂苷Rg3组细胞使用终质量浓度为 15.0 mg·L-1人参皂苷Rg3干预，奥希替尼组细胞使用0.1 μmol·L-1奥希替尼干预，奥希替尼+人参皂苷Rg3组细胞使用0.1 μmol·L-1奥希替尼和终质量浓度为15.0 mg·L-1人参皂苷Rg3干预，对照组细胞加入等量的磷酸盐缓冲溶液。采用Hoechst33342染料染色各组细胞，观察药物干预48 h时各组细胞形态变化；采用甲基噻唑基四唑法检测药物干预24、48、72 h时各组细胞增殖抑制率，采用流式细胞术检测药物干预48 h时各组细胞凋亡和细胞周期分布情况，采用实时荧光定量聚合酶链式反应检测药物干预48 h时各组细胞中磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（Akt）、caspase-3 mRNA相对表达量，采用Western blot法检测药物干预48 h时各组细胞中PI3K、Akt、caspase-3、磷酸化PI3K(p-PI3K)、磷酸化Akt（p-Akt）蛋白相对表达量。结果　药物干预48 h时，对照组细胞形态正常，人参皂苷Rg3组、奥希替尼组、奥希替尼+人参皂苷Rg3组细胞均出现核固缩、染色质凝集、核碎裂，部分出现凋亡小体，其中奥希替尼+人参皂苷Rg3组细胞形态变化最为明显，奥希替尼组次之，人参皂苷Rg3组细胞形态变化最轻。药物干预24、48、72 h时，人参皂苷Rg3组、奥希替尼组、奥希替尼+人参皂苷Rg3组细胞增殖抑制率均显著高于对照组（P<0.05）。人参皂苷Rg3组、奥希替尼组、奥希替尼+人参皂苷Rg3组细胞增殖抑制率随干预时间延长均显著升高，各时间点间增殖抑制率差异均有统计学意义（P<0.05）。药物干预48 h时，各药物干预组细胞凋亡率显著高于对照组（P<0.05）；奥希替尼组和奥希替尼+人参皂苷Rg3组细胞凋亡率显著高于人参皂苷Rg3组（P<0.05）；奥希替尼+人参皂苷Rg3组细胞凋亡率显著高于奥希替尼组（P<0.05）。药物干预48 h时，与对照组相比，人参皂苷Rg3组G0/G1期细胞占比升高，G2期细胞占比降低（P<0.05）；奥希替尼组和奥希替尼+人参皂苷Rg3组G0/G1期、G2期细胞占比均降低，S期细胞占比均升高（P<0.05）;与人参皂苷Rg3组相比，奥希替尼组和奥希替尼+人参皂苷Rg3组G0/G1期、G2细胞占比均降低，S期细胞占比均升高（P<0.05）;奥希替尼+人参皂苷Rg3组G2期细胞占比显著低于奥希替尼组（P<0.05）。药物干预48 h时，人参皂苷Rg3组、奥希替尼组、奥希替尼+人参皂苷Rg3组细胞中caspase-3 mRNA相对表达量均显著高于对照组（P<0.05），奥希替尼组和奥希替尼+人参皂苷Rg3组细胞中caspase-3 mRNA相对表达量均显著高于人参皂苷Rg3组（P<0.05），奥希替尼+人参皂苷Rg3组细胞中caspase-3 mRNA相对表达量显著高于奥希替尼组（P<0.05）；各组细胞中PI3K、Akt mRNA相对表达量比较差异均无统计学意义（P>0.05）。药物干预48 h时，人参皂苷Rg3组、奥希替尼组和奥希替尼+人参皂苷Rg3组细胞中caspase-3蛋白相对表达量均显著高于对照组（P<0.05），p-PI3K/PI3K、p-Akt/Akt均显著低于对照组（P<0.05）；奥希替尼组和奥希替尼+人参皂苷Rg3组细胞中caspase-3蛋白相对表达量均显著高于人参皂苷Rg3组，p-PI3K/PI3K、p-Akt/Akt均显著低于人参皂苷Rg3组（P<0.05）；奥希替尼+人参皂苷Rg3组中caspase-3蛋白相对表达量显著高于奥希替尼组（P<0.05），p-PI3K/PI3K、p-Akt/Akt显著低于奥希替尼组（P<0.05）。结论　奥希替尼联合人参皂苷Rg3可有效抑制非小细胞肺癌H1975细胞增殖，促进其凋亡，其机制可能与抑制PI3K/Akt信号通路有关。
Abstract:
Objective　To explore the effect of osimertinib and ginsenoside Rg3 on the proliferation and apoptosis of non-small cell lung cancer cell line H1975 and its mechnism.Methods　The H1975 cells in logarithmic growth phase were randomly divided into the ginsenoside Rg3 group,osimertinib group,osimertinib+ginsenoside Rg3 group and control group,with 5 multiple holes in each group.The cells in the ginsenoside Rg3 group were treated with ginsenoside Rg3 at a final concentration of 15.0 mg·L-1,the cells in the osimertinib group were treated with osimertinib at a final concentration of 0.1 μmol·L-1,the cells in the osimertinib+ginsenoside Rg3 group were treated with osimertinib with a final concentration of 0.1 μmol·L-1 and ginsenoside Rg3 with a final concentration of 0.1 μmol·L-1,and the cells in the control group were treated with an equal amount of phosphate buffered solution.The cells in each group were stained with Hoechst 33342 dye,and the morphological changes of the cells in each group were observed at 48 h after drug intervention；The methyl thiazolyl tetrazolium method was used to detect the inhibition rate of cell proliferation in each group at 24,48 and 72 h after drug intervention；The flow cytometry was used to detect the cell apoptosis and cell cycle distribution in each group at 48 h after drug intervention；The real-time fluorescence quantitative polymerase chain reaction was used to detect the relative expression of phosphatidylinositol 3-kinase (PI3K),protein kinase B (Akt) and caspase-3 mRNA in each group after drug intervention for 48 h；The western blot was used to detect the relative expression levels of PI3K,Akt,caspase-3,phosphorylated PI3K(p-PI3K) and phosphorylated Akt(p-Akt) protein of cells in each group after drug intervention for 48 h.Results　After 48 h of drug intervention,the morphology of the cells in the control group were normal.The cells in the ginsenoside Rg3 group,the osimertinib group and the osimertinib+ginsenoside Rg3 group all appeared nuclear pyknosis,chromatin agglutination,nuclear fragmentation,and some of them appeared apoptotic bodies.The cell morphology changes in the osimertinib+ginsenoside Rg3 group were the most obvious,followed by the osimertinib group,and the cells in the ginsenoside Rg3 group had the least changes.At 24,48,and 72 h of drug intervention,the cell proliferation inhibition rates in the ginsenoside Rg3 group,osimertinib group,osimertinib+ginsenoside Rg3 group were significantly higher than that in the control group (P<0.05).The cell proliferation inhibition rates in the ginsenoside Rg3 group,osimertinib group,osimertinib+ginsenoside Rg3 group increased significantly with the intervention time,and the differences in the proli-feration inhibition rates between each time point were statistically significant (P<0.05 ).At 48 h of drug intervention,the apoptosis rates in each drug intervention group were significantly higher than that in the control group (P<0.05);the apoptosis rates in the osimertinib group and osimertinib+ginsenoside Rg3 group were significantly higher than that in the ginseno-side Rg3 group (P<0.05);the apoptosis rate in the osimertinib+ginsenoside Rg3 group was significantly higher than that in the osimertinib group (P<0.05).After 48 h of drug intervention,compared with the control group,the proportion of G0/G1 phase cells in the ginsenoside Rg3 group increased,and the proportion of G2 phase cells decreased (P<0.05);the proportions of G0/G1 cells and G2 phases cells in the osimertinib group and osimertinib + ginsenoside Rg3 decreased,and the proportion of cells in S phase increased (P<0.05);compared with the ginsenoside Rg3 group,the proportions of G0/G1 and G2 phase cells in the osimertinib group and the osimertinib + ginsenoside Rg3 decreased,and the proportions of S phase cells increased (P<0.05);the proportion of cells in the G2 phase of the osimertinib + ginsenoside Rg3 group was significantly lower than that in the osimertinib group (P<0.05).Conclusion　Osimertinib combined with ginsenoside Rg3 can effectively inhibit the proliferation and promote apoptosis of non-small cell lung cancer H1975 cells,and the mechanism may be related to PI3K/Akt signaling pathway.

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更新日期/Last Update: 2020-11-05