参芪扶正注射液对肺癌细胞凋亡及NCR1/NKp46 通路的影响
DOI:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

R734.2

基金项目:


Effect of Shenqi Fuzheng Injection on Apoptosis of Lung Cancer Cells and NCR1/ NKp46 Signaling Pathway
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:探讨参芪扶正注射液对肺癌细胞凋亡及负性调节区域因子1/自然杀伤因子蛋白46(NCR1/NKp46) 通路的影响。方法:将人肺癌LEWIS 细胞分为LEWIS 组、顺铂组、参芪扶正注射液低、高剂量组(低、高剂量组),以上各组每孔设6 个平行样。LEWIS 组取10 mL 肺癌LEWIS 细胞液(细胞浓度为5×106/mL) 于10%胎牛血清的DMEM 培养液中;顺铂组细胞培养方法同LEWIS 组,加入顺铂使终浓度为50.0 μg/mL;低、高剂量组细胞培养方法同LEWIS 组,分别加入参芪扶正注射液,使终浓度为50.0 μg/mL、100.0 μg/mL,培养72 h。培养结束后,MTT 法测定各组细胞活力,结晶紫染色测定单克隆形成数目,膜联蛋白V-异硫氰酸荧光素/碘化丙啶凋亡检测试剂盒检测细胞凋亡水平,RT-PCR 法及Western blot 法测定NCR1、NKp46 基因与蛋白表达水平。结果:与LEWIS 组比较,顺铂组与低、高剂量组细胞OD 值、存活率、克隆形成数目降低,凋亡率、G1 期水平及NCR1、NKp46 mRNA 和蛋白表达升高(P<0.05)。与顺铂组比较,低剂量组细胞OD 值、存活率、克隆形成数目升高,凋亡率、G1 期水平及NCR1、NKp46 mRNA 和蛋白表达降低(P<0.05);高剂量组细胞OD 值、存活率、克隆形成数目降低,凋亡率、G1 期水平及NCR1、NKp46 mRNA 和蛋白表达升高(P<0.05)。与低剂量组比较,高剂量组OD 值、存活率、细胞克隆形成数目降低(P<0.05),凋亡率、G1 期水平及细胞NCR1、NKp46 mRNA 和蛋白表达均升高(P<0.05)。结论:参芪扶正注射液能抑制人肺癌LEWIS 细胞增殖,促进其凋亡;其机制与参芪扶正注射液可促进肺癌LEWIS 细胞NCR1、NKp46 mRNA 与蛋白高表达,进而介导NK 细胞杀伤肺癌细胞有关。

    Abstract:

    Abstract:Objective:To discuss the effect of Shenqi Fuzheng injection on apoptosis of the lung cancer cells and Negative Control Region 1/Natural Killer-factor protein46(NCR1/NKp46) signaling pathway. Methods:Lewis cells of human lung cancer were divided into the LEWIS group,the cisplatin group,the Shenqi Fuzheng injection groups of low-dose and high-dose. There were six parallel samples in each hole in each group. In the LEWIS group,10 mL cell sap of lung cancer LEWIS(cell concentration was 5×106/mL) was placed in DMEM culture medium with 10% fetal bovine serum. The cell culture method in the cisplatin group was the same in the LEWIS group,and cisplatin was added to make the final concentration of 50.0 μg/mL. The cell culture methods in the Shenqi Fuzheng injection groups of low- dose and high- dose were the same in the LEWIS group;the two groups were added Shenqi Fuzheng injection accordingly,and the final concentration was 50.0 μg/mL and 100.0 μg/mL respectively;both groups were cultured for 72 hours. After cultivation,MTT method was applied for detecting cell viability; crystal violet staining was carried out to detect the number of monoclonal formation; annexin v- fluorescein isothiocyanate/apoptosis detection kit of propidium iodide was used for detect the level of apoptosis;PT- PCR method and Western blot method were applied for detecting expression levels of gene and protein of NCR1 and NKp46. Results: Compared with those in the LEWIS group,in the cisplatin group and the Shenqi Fuzheng injection groups of low-dose and high- dose,the cell OD value,survival rate and the number of colony formation were decreased,and the apoptosis rate, G1 period as well as expression levels of mRNA and protein of NCR1 and NKp46 were increased(P<0.05). Compared with those in the cisplatin group,in the low- dose group,the cell OD value,survival rate and the number of colony formation were increased,and the apoptosis rate,G1 period as well as expression levels of mRNA and protein of NCR1 and NKp46 were decreased(P<0.05);in the high-dose group,the cell OD value,survival rate and the number of colony formation were decreased,and the apoptosis rate,G1 period as well as expression levels of mRNA and protein of NCR1 and NKp46 were increased(P<0.05). Compared with those in the low-dose group,the cell OD value,survival rate and the number of colony formation in the high-dose group were decreased(P<0.05),and the apoptosis rate,G1 period as well as expression levels of mRNA and protein of NCR1 and NKp46 were increased(P<0.05). Conclusion: Shenqi Fuzheng injection can inhibit the proliferation of human lung cancer LEWIS,promote the apoptosis of LEWIS. Its mechanism is that Shenqi Fuzheng injection can promote the mRNA and protein expression of NCR1 and NKp46 in the lung cancer LEWIS cell,and then mediate NK cells to kill the lung cancer cells.

    参考文献
    相似文献
    引证文献
引用本文

徐婷婷,李隆祥,王晨,陈锋.参芪扶正注射液对肺癌细胞凋亡及NCR1/NKp46 通路的影响[J].新中医,2020,52(15):7-10

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2020-08-08
  • 出版日期: