凝聚素Ⅰ复合物亚基G促进小鼠胃腺癌发展、诱导免疫逃逸
NCAPG promotes the development of gastric adenocarcinoma and induces immune escape in mice
董跃华 1王大伟 1杨燕君 1魏玉磊 1高永山 1姜伟华 1王贵刚 1张振明1
作者信息
- 1. 河北北方学院附属第一医院胸心外科,河北张家口 075000
- 折叠
摘要
[目的]为研究凝聚素Ⅰ复合物亚基G(NCAPG)对胃腺癌发展及免疫逃逸相关因子的影响,通过干扰NCAPG后,观察胃腺癌AGS细胞的在体内外的变化,以期为胃腺癌的治疗提供新思路.[方法]将裸鼠和AGS细胞均分为3组:对照组、si-NC组和si-NCAPG组.采用CCK-8检测细胞增殖情况.采用实时定量PCR检测临床IL-2、IFN-γ、TNF-α和PD-L1 mRNA的表达水平.采用免疫组化法检测肿瘤组织中Ki-67的表达水平.采用流式技术检测NCAPG对AGS细胞凋亡的影响.通过蛋白免疫印迹实验检测PI3KAKT蛋白表达水平.[结果]与对照组和si-NC比较,si-NCAPG组的裸鼠体内肿瘤组织生长速度最慢,Bax/Bcl-2比值升高,Ki-67表达降低,IL-2、IFN-γ、TNF-α的mRNA水平升高,PD-L1的mRNA水平降低,PI3K和AKT蛋白的磷酸化水平降低(P＜0.05).与对照组和si-NC比较,si-NCAPG组的细胞增殖倍数最低、凋亡率最高,IL-2、IFN-γ、TNF-α的mRNA水平升高,PD-L1的mRNA水平降低,PI3K和AKT蛋白的磷酸化水平降低(P＜0.05).[结论]NCAPG可促进胃腺癌的发生及诱导免疫逃逸,其作用机制可能与PI3K/AKT信号通路相关.
Abstract
[Objective]To study the effect of clusterin Ⅰ complex subunit G(NCAPG)on the development of gastric adeno-carcinoma and immune escape related factors,we observed the changes of gastric adenocarcinoma AGS cells in vitro and in vivo after interfering with NCAPG,in order to provide new ideas for the treatment of gastric adenocarcinoma.[Method]Both nude mice and AGS cells were divided into 3 groups:control group,si-NC group and si-NCAPG group.CCK-8 was used to detect cell proliferation.The expression levels of clinical IL-2,IFN-γ,TNF-α and PD-L1 mRNA were detected by real-time quantitative PCR.The expression level of Ki-67 in tumor tissues was detected by immunohistochemistry.The effect of NCAPG on apoptosis of AGS cells was detected by flow-through technique.The expression levels of PI3KAKT proteins were detected by Western Blot.[Result]Compared with the control group and si-NC,The si-NCAPG group had the slowest tumor growth rate(466.56±15.02 vs 503.97±12.88 vs 203.78±10.22)and higher Bax/Bcl-2 ratio,Ki-67 expression decreased,IL-2,IFN-γ and TNF-α mRNA levels increased,and PD-L1 mRNA level decreased,phosphorylation of PI3K and AKT(P＜0.05).Compared with the control group and si-NC group,the cell proliferation was the lowest,the apoptosis rate was the highest,the mRNA levels of IL-2,IFN-γ and TNF-α were increased,the mRNA level of PD-L1 was decreased,and the phosphorylation levels of PI3 K and AKT protein were decreased in the si-NCAPG group(P＜0.05).[Conclusion]NCAPG promotes gastric adenocarcinoma and induces immune escape,and its mechanism of action may be related to the PI3K/AKT signaling pathway.
关键词
凝聚素Ⅰ复合物亚基G/PI3K/AKT/增殖/胃腺癌/免疫逃逸/凋亡/裸鼠Key words
clusterin Ⅰ complex subunit G/PI3K/AKT/proliferation/gastric adenocarcinoma/immune escape/apoptosis/nude mice引用本文复制引用
基金项目
河北省卫生计生委项目([2017]46号)
出版年
2024
NETL
NSTL
万方数据