miR-221-3p靶向FGF14调控肺腺癌细胞增殖、凋亡、迁移及侵袭的分子机制  

Molecular mechanism of miR-221-3p targeting FGF14 on regulating proliferation,apoptosis,migration and invasion of lung adenocarcinoma cells

作　　者：李淑勤 赵树强[2] 郑丽琴[3] LI Shuqin;ZHAO Shuqiang;ZHENG Liqin(Department of Internal Medicine,Department of Clinical Medicine,Fenyang College of Shanxi Medical University,Lyuliang 032200,China)

机构地区：[1]山西医科大学汾阳学院临床医学系内科教研室,吕梁032200 [2]山西医科大学附属汾阳医院检验科,吕梁032200 [3]山西医科大学附属汾阳医院呼吸科,吕梁032200

出　　处：《中国免疫学杂志》2022年第7期827-832,共6页Chinese Journal of Immunology

基　　金：山西省卫生计生委科研课题项目(201602051)。

摘　　要：目的:探讨miR-221-3p是否通过靶向FGF14调节肺腺癌细胞增殖、凋亡、迁移及侵袭的生物学行为。方法:采用qRT-PCR法与Western blot法分别检测人支气管上皮细胞(BEAS-2B)、人肺腺癌细胞(A549、A-427、PC-9)中miR-221-3p、FGF14的表达水平;分别将miR-inhibitor-NC、miR-221-3p inhibitor、si-NC、si-FGF14、miR-inhibitor-NC+si-NC、miR-221-3p inhibotor+si-NC、miR-221-3p inhibotor+si-FGF14转染至PC-9细胞;采用MTT实验检测细胞增殖;采用Transwell小室实验检测细胞迁移及侵袭;采用流式细胞术检测细胞凋亡率;双荧光素酶报告实验检测miR-221-3p、FGF14靶向关系;Western blot法检测CyclinD1、MMP-2、MMP-9、Bcl-2、P21、Bax蛋白表达量。结果:与BEAS-2B细胞比较,肺腺癌细胞A549、A-427、PC-9中miR-221-3p的表达水平升高(P<0.05),FGF14 mRNA及蛋白水平降低(P<0.05);转染miR-221-3p inhibitor可明显降低OD值及CyclinD1、MMP-2、MMP-9、Bcl-2蛋白水平(P<0.05),提高凋亡率及P21、Bax蛋白水平(P<0.05),减少迁移及侵袭细胞数(P<0.05);双荧光素酶报告实验证实FGF14是miR-221-3p的靶基因;转染si-FGF14可提高OD值及CyclinD1、MMP-2、MMP-9、Bcl-2蛋白水平(P<0.05),增加迁移及侵袭细胞数(P<0.05),降低凋亡率及P21、Bax蛋白水平(P<0.05);转染si-FGF14可明显逆转转染miR-221-3p inhibitor对PC-9细胞增殖、凋亡、迁移及侵袭的作用。结论:抑制miR-221-3p表达可通过上调FGF14的表达从而抑制肺腺癌细胞增殖、迁移、侵袭及诱导细胞凋亡。Objective:To explore whether miR-221-3p regulated the biological behavior of lung adenocarcinoma cell proliferation,apoptosis,migration and invasion by targeting FGF14.Methods:qRT-PCR and Western blot were used to detect the expression levels of miR-221-3p and FGF14 in human bronchial epithelial cells(BEAS-2B)and human lung adenocarcinoma cells(A549,A-427,PC-9).miR-inhibitor-NC,miR-221-3p inhibitor,si-NC,si-FGF14,miR-inhibitor-NC+si-NC,miR-221-3p inhibotor+siNC,miR-221-3p inhibotor+si-FGF14 were transfected into PC-9 cells.MTT assay was used to detect cell proliferation.The Transwell chamber experiment was used to detect cell migration and invasion.Flow cytometry was used to detect the apoptosis rate.The dual luciferase reporter experiment was used to detect the targeting relationship of miR-221-3p and FGF14.Western blot was used to detect the protein expression of CyclinD1,MMP-2,MMP-9,Bcl-2,P21,Bax.Results:Compared with BEAS-2B cells,the expression levels of miR-221-3p in lung adenocarcinoma cells A549,A-427,and PC-9 were increased(P<0.05),while the level of mRNA and the protein level of FGF14 were decreased(P<0.05).Transfection of miR-221-3p inhibitor could significantly reduce the OD value and the protein levels of CyclinD1,MMP-2,MMP-9 and Bcl-2(P<0.05),and increased the apoptosis rate and the protein levels of P21 and Bax(P<0.05),reduced the number of migration and invasion cells(P<0.05).The dual luciferase reporter experiment confirmed that FGF14 was the target gene of miR-221-3p.Transfection of si-FGF14 could increase the OD value and the protein levels of CyclinD1,MMP-2,MMP-9 and Bcl-2(P<0.05),increased the number of migration and invasion cells(P<0.05),and reduced the rate of apoptosis and the protein levels of P21,Bax(P<0.05).Transfection of si-FGF14 could significantly reversed the effects of transfection of miR-221-3p inhibitor on the proliferation,apoptosis,migration and invasion of PC-9 cells.Conclusion:Inhibition of miR-221-3p expression could inhibit lung adenocarcinoma cell proliferation,migration

关 键 词：miR-221-3p FGF14 肺腺癌 增殖 凋亡 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]