实用医学杂志 ›› 2020, Vol. 36 ›› Issue (21): 2916-2920.doi: 10.3969/j.issn.1006⁃5725.2020.21.006

• 基础研究 • 上一篇    下一篇

成纤维细胞活化蛋白对眼睑基底细胞癌相关成纤维细胞增殖及凋亡的影响及机制研究

杨夏,张西,龙秋蓉   

  1. 贵州医科大学附属医院眼科(贵阳550004)
  • 出版日期:2020-11-10 发布日期:2020-11-30
  • 基金资助:
    贵阳市科技局基金(编号:2010 筑科农合同字第1-社-18号)

Effects of FAP on proliferation and apoptosis of fibroblasts associated with eyelid basal cell carcinoma and its mechanism

YANG Xia,ZHANG Xi,LONG Qiurong   

  1. Department of Ophthalmology,Affiliated Hospital of Gui⁃zhou Medical University,Guiyang 550004,China
  • Online:2020-11-10 Published:2020-11-30

摘要:

目的 观察成纤维细胞活化蛋白(fibroblast activation protein,FAP)对眼睑基底细胞癌相关成纤维细胞(carcinoma associated fibroblasts,CAFs)增殖、凋亡的影响。方法 分离培养鉴定眼睑CAFs 和正常成纤维细胞(normal fibroblasts,NFs)。CAFs随机分为对照组、过表达组(FAP)、敲降组(FAP shRNA)、空载组(随机shRNA)、抑制剂组[FAP shRNA+第10号染色体上磷酸酶和张力蛋白同源缺失的基因(PTEN)抑制剂bpv],MTT法观察增殖,流式细胞术检测凋亡,Western blot法检测观察眼睑CAFs FAP、PTEN、磷脂酰肌醇3 激酶(PI3K)、p⁃PI3K、丝氨酸⁃苏氨酸蛋白激酶(Akt)、p⁃Akt 蛋白。结果 培养并鉴定眼睑CAFs 和NFs。与对照组、空载组比较,过表达组吸光度(A)值增加,凋亡率降低,FAP、p⁃PI3K、p⁃Akt 表达升高,PTEN 表达降低,敲降组、抑制剂组A 值减小,凋亡率升高,FAP、p⁃PI3K、p⁃Akt 表达降低,PTEN 表达升高(P<0.05);抑制剂组A 值、p⁃PI3K、p⁃Akt 表达高于敲降组、凋亡率、PTEN 表达低于敲降组(P<0.05)。结论 过表达FAP可促进CAFs增殖,抑制其凋亡,可能是通过抑制PTEN,激活PI3K/AKT通路发挥作用。

关键词: 眼睑基底细胞癌, 成纤维细胞活化蛋白, 增殖, 凋亡, PI3K/AKT通路

Abstract:

Objective To observe the effect of fibroblast activation protein(FAP)on the proliferationand apoptosis of eyelid basal cell carcinoma⁃associated fibroblasts(CAFs). Methods Eyelid CAFs and normalfibroblasts(NFs)were isolated,cultured and identified. CAFs were randomly divided into control group,overex⁃pression group(FAP),knockdown group(FAP shRNA),empty group(random shRNA)and inhibitor group[FAP shRNA+bpv]. MTT method was used to observe the proliferation;flow cytometry to detect apoptosis andwestern blot method to detect eyelid CAFs FAP,PTEN,phosphatidyl inositol⁃3⁃kinase(PI3K),p⁃PI3K,serine⁃threonine protein kinase(Akt)and p⁃Akt protein. Results Eyelid CAFs and NFs were cultured and identified.Compared with those of the control group and empty group,the absorbance(A)value of the overexpression groupincreased;apoptosis rate decreased;FAP,p⁃PI3K and p⁃Akt protein increased and PTEN protein decreased. Avalue of the knockdown and inhibitor group decreased;apoptosis rates increased;FAP,p⁃PI3K,and p⁃Akt pro⁃tein decreased and PTEN protein increased(P < 0.05). A value,p⁃PI3K and p⁃Akt protein of the inhibitor groupwere higher than those in the knockdown group,but the rate of apoptosis and PTEN protein were lower(P < 0.05).Conclusion Overexpression of FAP can promote the proliferation of CAFs and inhibit their apoptosis,possibly byinhibiting PTEN and activating the PI3K/AKT pathway.


Key words: eyelid basal cell carcinoma, fibroblast activation protein, proliferation, apoptosis, PI3K/AKT pathway