摘要
目的探讨环状RNA(circRNA)circZFR在吉非替尼抑制肺腺癌细胞生长中的作用。方法将PC-9/ZD细胞分为对照组、吉非替尼组、吉非替尼+si-con组和吉非替尼+si-circZFR组。实时荧光定量PCR(qRT-PCR)检测circZFR、微小RNA(miR)-1236在肺腺癌组织或肺腺癌细胞中的表达水平,细胞计数试剂盒-8(CCK-8)检测细胞抑制率、克隆形成实验检测克隆形成数,流式细胞术检测细胞凋亡率,原位末端标记法(TUNEL)检测细胞凋亡指数,蛋白免疫印迹(Western blot)检测细胞周期蛋白D1(CyclinD1)、增殖细胞核抗原(PCNA)、B淋巴细胞瘤-2相关X蛋白(Bax)和B淋巴细胞瘤-2(Bcl-2)蛋白表达水平,双荧光素酶实验检测circZFR与miR-1236的靶向关系。结果CircZFR在吉非替尼不敏感肺腺癌组织和细胞中高表达。MiR-1236在吉非替尼不敏感肺腺癌组织和细胞中低表达。与对照组比较,吉非替尼组PC-9/ZD细胞circZFR表达水平、细胞抑制率、凋亡率、凋亡指数和Bax蛋白表达水平明显增加(t值分别为-12.471、-15.305、-8.338、-13.544和-5.724,P<0.05),miR-1236表达水平、克隆形成数、CyclinD1、PCNA和Bcl-2蛋白表达水平明显降低(t值分别为27.429、4.276、3.882、6.379和5.982,P<0.05),差异有统计学意义。与吉非替尼+si-con组比较,吉非替尼+si-circZFR组PC-9/ZD细胞miR-1236表达水平、细胞抑制率、凋亡率、凋亡指数和Bax蛋白表达水平明显增加(t值分别为-7.588、-7.515、-11.546、-9.610和-4.473,P<0.05),circZFR表达水平、克隆形成数、CyclinD1、PCNA和Bcl-2蛋白表达水平明显降低(t值分别为8.444、4.356、6.688、7.298和5.824,P<0.05),差异有统计学意义。circZFR靶向调控miR-1236的表达。结论沉默circZFR可增强吉非替尼对肺腺癌细胞的抑制作用。
Objective To investigate the role of circular RNA(circRNA)circZFR in gefitinib inhibiting the growth of lung adenocarcinoma cells.Methods PC-9/ZD cells were divided into control group,gefitinib group,gefitinib+si-con group and gefitinib+si-circZFR group.Real-time quantitative reverse transcriptionPCR(qRT-PCR)was used to detect the expression levels of circZFR and microRNA(miR)-1236 in lung adenocarcinoma tissues or lung adenocarcinoma cells.Cell counting kit-8(CCK-8)was used to detect cell inhibition rate.Clone formation assay was used to detect the number of clone formation.Flow cytometry was used to detect cell apoptosis rate.In situ end labeling(TUNEL)was used to detect apoptosis index.Western blot was used to detect the protein expression levels of CyclinD1,proliferating cell nuclear antigen(PCNA),B-lymphocytoma-2-associated X protein(Bax)and B-lymphocytoma-2(Bcl-2).Dual luciferase assay was used to detect the targeting relationship between circZFR and miR-1236.Results CircZFR was highly expressed in lung adenocarcinoma tissues and cells insensitive to gefitinib.MiR-1236 was low expressed in lung adenocarcinoma tissues and cells insensitive to gefitinib.Compared with the control group,the circZFR expression level,cell inhibition rate,apoptosis rate,apoptosis index and Bax protein expression level of PC-9/ZD cells in gefeitinib group were significantly increased(t values were-12.471,-15.305,-8.338,-13.544,-5.724 respectively,P<0.05),while the miR-1236 expression level,number of clone formation,protein expression levels of CyclinD1,PCNA,and Bcl-2 were significantly decreased(t values were 27.429,4.276,3.882,6.379,5.982 respectively,P<0.05).The difference was statistically significant.Compared with the gefitinib+si-con group,the miR-1236 expression level,cell inhibition rate,apoptosis rate,apoptosis index and Bax protein expression level of PC-9/ZD cells in gefitinib+si-circZFR group were significantly increased(t values were-7.588,-7.515,-11.546,-9.610,-4.473,respectively,P<0.05),while the circZFR expression le
作者
施晓宁
沈云珠
庄奕筠
黄培瑜
SHI Xiao-ning;SHEN Yun-zhu;ZHUANG Yi-jun;HUANG Pei-yu(The Second Affiliated Hospital of Fujian Medical University,Quanzhou Fujian 362000,China)
出处
《毒理学杂志》
CAS
2023年第3期223-229,共7页
Journal of Toxicology
基金
2019年福建医科大学启航基金项目计划(2019QH1127)。
