天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (9): 807-812.doi: 10.11958/20200157

• 细胞与分子生物学 • 上一篇    下一篇

甲磺酸阿帕替尼通过p53抑制结肠癌细胞HCT116增殖的作用及机制

杨瀚林 1，汤弘婷 1，杨娟 1，刘虹麟 2，李梦醒 3，李琴山 1，4△
  

  1. 1贵州医科大学医学检验学院（邮编550004）；2中日友好医院临床医学研究所；3贵州医科大学附属医院血液科；4贵州省产前诊断中心
  • 收稿日期:2020-01-13 修回日期:2020-05-22 出版日期:2020-09-15 发布日期:2020-09-15
  • 通讯作者: 李琴山 E-mail:qinshanli@hotmail.com
  • 作者简介:杨瀚林（1993），男，硕士在读，主要从事结肠癌发病机制及治疗研究
  • 基金资助:
    国家自然科学基金资助项目（81460365，81402451，81760039，81960476）；贵州省科技厅资助项目（No.2019-1270）；贵州医科大
    学附属医院博士启动基金资助项目（    201404-001）；贵阳市科技局资助项目（筑科合同20151001-社33号-J-2015-09）

Effects and mechanism of apatinib on the proliferation of HCT116 inhibited by p53

YANG Han-lin1, TANG Hong-ting1, YANG Juan1, LIU Hong-lin2, LI Meng-xing3, LI Qin-shan1, 4△ #br#   

  1. 1 School of Medical Laboratory Science, Guizhou Medical University, Guiyang 550004, China; 2 Institute of Clinical Medical
    Sciences, China-Japan Friendship Hospital; 3 Department of Hematology, the Affiliated Hospital of Guizhou Medical
    University; 4 Guizhou Provincial Prenatal Diagnosis Center
  • Received:2020-01-13 Revised:2020-05-22 Published:2020-09-15 Online:2020-09-15

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摘要： 摘要：目的 探讨阿帕替尼对人结肠癌细胞HCT116及敲除野生型p53的HCT116细胞抑制作用的差异并探讨
其机制。方法 以0、15、30、60 μmol/L阿帕替尼处理HCT116 p53+/+、HCT116 p53-/-细胞24、48 h，以CCK-8法检测阿
帕替尼对两株细胞增殖的抑制作用；流式细胞术（Annexin V/PI法）检测0、15、30 μmol/L阿帕替尼处理细胞24 h后凋
亡率变化；实时荧光定量PCR法和Western blot法检测0、15、30 μmol/L阿帕替尼处理细胞24 h后，p53、NF-κB p65、
Caspase-3 mRNA和蛋白表达变化。结果 CCK-8结果显示，阿帕替尼能抑制HCT116 p53+/+、HCT116 p53-/-细胞的增
殖，抑制作用具有剂量依赖性（P＜0.01）。流式细胞术结果显示，阿帕替尼干预后，HCT116 p53+/+、HCT116 p53-/-细胞
凋亡率升高（P＜0.01）。与HCT116 p53+/+细胞相比，阿帕替尼处理后HCT116 p53-/-细胞的凋亡率较低（P＜0.05）。阿
帕替尼处理后，HCT116 p53+/+、HCT116 p53-/-细胞中 p53、NF-κB p65、capsase-3 mRNA 和 Caspase-3 蛋白表达升高
（P＜0.05）。阿帕替尼干预后，HCT116 p53+/+细胞中 p53、NF-κB p65 蛋白表达下调而 HCT116 p53-/-细胞中 NF-κB
p65蛋白表达上调（P＜0.01）。结论 阿帕替尼可能通过p53/NF-κB通路抑制HCT116 p53+/+、HCT116 p53-/-细胞增殖
并促进细胞凋亡，结肠癌细胞可能通过野生型p53对阿帕替尼产生耐药。

关键词: 结肠肿瘤；HCT116细胞；基因, p53；NF-κB；转录因子RelA；细胞增殖；细胞凋亡；阿帕替尼

Abstract: Abstract: Objective To investigate the inhibition effects of apatinib on human colon cancer cells HCT116 p53+/+and
HCT116 p53-/-, and further explore the molecular mechanisms in vitro. Methods The gradient concentrations of apatinib
(0, 15,30 and 60 μmol/L) were used to treat HCT116 p53+/+ and HCT116 p53-/- for 24 h and 48 h. The inhibitory effects of
apatinib on the proliferation of two cell lines were detected by CCK-8. Flow cytometry (Annexin V/PI method) was used to
detect the apoptosis rates of apatinib treated HCT116 p53+/+ and HCT116 p53-/-cells. The expression levels of p53, NF-κB
p65, Caspase-3 mRNA and protein of apatinib treated HCT116 p53+/+ and HCT116 p53-/-cells were revealed by real-time
PCR and Western blot assay. Results CCK-8 results showed that apatinib could inhibit the viability of HCT116 p53+/+ and
HCT116 p53-/-cells in a dose-dependent manner (P＜0.01). Flow cytometry results showed that after treatment with
apatinib, the apoptosis rates of HCT116 p53+/+ and HCT116 p53-/-cells were increased (P＜0.01). Compared with HCT116
p53+/+ , the pro-apoptotic effect of apatinib on HCT116 p53-/- was lower (P＜0.05). After treatment with apatinib, the
expression levels of p53, NF- κB p65, capsase-3 mRNA and Caspase-3 protein were increased in HCT116 p53+/+ and
HCT116 p53-/-cells (P＜0.05). After treatment with apatinib, the protein expressions of p53 and NF- κB p65 were downregulated in HCT116 p53+/+ cells, while the protein expression of NF-κB p65 was up-regulated in HCT116 p53-/- cells (P＜
0.01). Conclusion Apatinib could inhibit cell proliferation and promote the apoptosis of HCT116 p53+/+ and HCT116
p53-/- cells through p53/NF-κB pathway. Colorectal cancer cells may develop drug resistance to apatinib through HCT116
p53-/- cells.

Key words: colonic neoplasms;HCT116 cells, genes, p53, NF-kappa B, transcription factor RelA, cell proliferation, apoptosis, apatinib

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