摘要
目的研究沉默纤维蛋白原样蛋白1(FGL1)对肺腺癌细胞吉非替尼药物敏感性的影响。方法实时荧光定量PCR及Western blot法检测FGL1在肺腺癌吉非替尼敏感细胞株PC9与吉非替尼耐药细胞株PC9/GR中差异性表达;然后在吉非替尼耐药株PC9/GR中沉默FGL1表达;CCK-8实验检测各组细胞增殖情况;流式细胞术测定各组细胞总凋亡率;Western blot实验测定各组凋亡相关蛋白Bcl-2、Bax、Caspase-3及Cleaved-Caspase-3的表达水平。结果FGL1在PC9/GR细胞中呈高表达;沉默FGL1可抑制PC9/GR的增殖,且提高了PC9/GR细胞凋亡率(P<0.05);Western blot表明,Caspase-3在各组细胞中表达差异无统计学意义(P>0.05);与PC9/GR组比较,吉非替尼组Bax、Bcl-2表达无明显差异,而Cleaved-Caspase-3表达上调,差异有统计学意义(P<0.05)。沉默FGL1后,Bcl-2蛋白表达下调,Bax、Cleaved-Caspase-3蛋白表达上调,差异有统计学意义(P<0.05)。结论沉默FGL1可增强肺腺癌耐药细胞对吉非替尼的敏感性,抑制细胞增殖,促进细胞凋亡,FGL1可能是治疗表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs)耐药性患者的一个潜在靶点。
Objective To investigate the effect of silenced fibrin original protein 1(FGL1)on the drug sensitivity of gefitinib in lung adenocarcinoma cells.Methods Real-time quantitative PCR and Western blot were used to detect the differential expression of FGL1 in lung adenocarcinoma gefitinib-sensitive cell line PC9 and gefitinib-resistant cell line PC9/GR.FGL1 expression was silenced in gefitinib resistant strain PC9/GR.CCK-8 assay was used to detect cell proliferation in each group.Flow cytometry was used to determine the apoptosis rate of each group.Western blot was used to determine relative expression levels of apoptotic proteins Bcl-2,Bax,Caspase-3 and Cleaved-Caspase-3.Results FGL1 was highly expressed in PC9/GR cells.Silencing FGL1 inhibited the proliferation of PC9/GR and increased the apoptosis rate of PC9/GR cells(P<0.05).Western blot showed no significant difference in the expression of Caspase-3 in each group(P>0.05).Compared with the control group,the expression of Bax and Bcl-2 in gefitinib group showed no significant difference,while the expression of Cleaved-Caspase-3 was up-regulated,with statistically significant difference(P<0.05).After FGL1 was silenced,Bcl-2 protein expression was down-regulated,while Bax and Cleaved-Caspase-3 protein expression was up-regulated,with statistically significant differences(P<0.05).Conclusion Silencing FGL1 can inscrease the sensitivity of lung adenocarcinoma cells to gefitinib,inhibit cell proliferation,promote cell apoptosis,and provide potential therapeutic targets for gefitinib resistant lung adenocarcinoma patients.
作者
高薇薇
孙翠兰
郝吉庆
Gao Weiwei;Sun Cuilan;Hao Jiqing(Dept of Oncology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第6期854-859,共6页
Acta Universitatis Medicinalis Anhui
基金
安徽省2018年度重点研究与开发计划项目(编号:1804h08020240)。