环状RNA含有16A的脱水酶结构域分子-微小RNA-1237-5p-细胞凋亡相关酪氨酸激酶轴调控乳腺癌细胞密歇根癌症基金会7号细胞凋亡  

CircularRNA abhydrolase domain containing 16A-microRNA-1237-5p-apoptosis associated tyrosine kinase axis regulates the apoptosis of michigan cancer foundation-7 in breast cancer cells

作　　者：盛志娟[1] 姜专基[1] 张斌明[1] 宋波[1] Sheng Zhijuan;Jiang Zhuanji;Zhang Binning;Song Bo(Department of Breast Surgery,Gansu Cancer Hospital,Gansu 730050,China)

机构地区：[1]甘肃省肿瘤医院乳腺外科,730050

出　　处：《中华实验外科杂志》2019年第12期2176-2178,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察环状RNA含有16A的脱水酶结构域分子(circRNA ABHD16A)-微小RNA(miRNA,miR)-1237-5p-细胞凋亡相关酪氨酸激酶(AATK)轴对乳腺癌细胞密歇根癌症基金会7号细胞(MCF-7)凋亡的影响.方法运用中国科学引文数据库(CSCD)和Target Scan在线分析分别分析ABHD16A与miR-1237-5p和miR-1237-5p与AATK的相关性.利用Luciferase assay检测circRNA ABHD16A是否靶向miR-1237-5p以及miR-1237-5p是否靶向AATK.在过表达或敲低circRNA ABHD16A的情况下,通过蛋白质印迹法(Western blot)检测AATK和乳腺癌细胞MCF-7凋亡标志蛋白表达;通过噻唑蓝(MTT)实验分析乳腺癌细胞MCF-7的细胞活力;通过磷脂酰丝氨酸外翻分析(Annexin V)染色检测乳腺癌细胞MCF-7的凋亡水平.Student's t检验分析两组单向方差分析.结果circRNA ABHD16A靶向miR-1237-5p;miR-1237-5p靶向AATK3'UTR的320-326和469-475区域.过表达ABHD16A时,AATK的表达量明显上升(t=15.295,P<0.01),细胞凋亡相关蛋白bcl-2拮抗剂(bak),bcl-2相关X凋亡调节因子(bax)和被切割的胱天蛋白酶9(cleaved-Cas9)的表达量明显上升(t=7.403、8.381、21.583,P<0.01),而bcl-2细胞凋亡调节剂(bcl-2)的表达量明显下降(t=6.301,P<0.01),MCF7细胞活力水平下降(t=17.392,P<0.01),凋亡水平上升(t=8.491,P<0.05).敲低ABHD16A时,AATK的表达量明显下降(t=9.290,P<0.01),细胞凋亡相关蛋白bak、bax和cleaved-Cas9的表达量明显下降(t=8.381、5.296、11.492,P<0.01),而bcl-2的表达量明显上升(t=5.609,P<0.01),MCF7细胞活力水平上升(t=5.694,P<0.01),凋亡水平下降(t=13.502,P<0.01).结论circRNA ABHD16A靶向miR-1237-5p后增加AATK的表达量,并促进乳腺癌细胞MCF-7凋亡.Objective To investigate the circular RNA abhydrolase domain containing 16A(circRNA ABHD16A)-microRNA(miRNA,miR)-1237-5p-Apoptosis associated tyrosine kinase(AATK)axis on apoptosis of breast cancer cell munggan cancer foundation-7(MCF-7).Methods The correlation between ABHD16A and miR-1237-5p and miR-1237-5p and AATK was analyzed by CSCD and TargetScan online analysis.The luciferase assay was used to detect whether circRNA ABHD16A targets miR-1237-5p and whether miR-1237-5p targets AATK.The expression of apoptotic marker proteins in AATK and breast cancer cells MCF-7 was detected by Western blot after over-expression or knockdown of circRNA ABHD16A.Cell viability of breast cancer cells MCF-7 was analyzed by methyl thiazolyl tetrazolium(MTT)assay;staining by Annexin-V was performed by MTT assay.The level of apoptosis of breast cancer cells MCF-7 was measured.Student'st-test analyzed two sets of one-way analysis of variance.Results circRNA ABHD16A targets miR-1237-5p;miR-1237-5p targets the 320-326 and 469-475 regions of the AATK 3'UTR.When overexpressing ABHD16A,AATK expression increased significantly(t=15.295,P<0.01),apoptosis-related protein bcl-2 antagonist/killer 1(bak),bcl-2 Associated X Apoptosis Regulator(bax)and cleaved Caspase-9(cleaved-CAS9)expression levels increased significantly(Z=7.403,&381,21.583,P<0.01).The expression of bcl-2 Apoptosis Regulator(bcl-2)was significantly decreased(t=6.301,P<0.01),the activity of MCF7 cells was decreased(t=17.392,P<0.01),and the level of apoptosis was increased(t=8.491,P<0.05).When ABHD16A was knocked down,the expression of AATK was significantly decreased(t=9.290,P<0.01),and the expression levels of apoptosis-related proteins bak,bax and cleaved-CAS9 were significantly decreased t=8.381,5.296,11.492,P<0.01),while the expression of bcl-2 increased significantly(t=5.609,P<0.01),the activity level of MCF7 cells increased(t=5.694,P<0.01),and the level of apoptosis decreased(t=13.502,P<0.01).Conclusion circRNA ABHD16A targets miR-1237-5p and increases the expression of A

关 键 词：环状RNA含有16A的脱水酶结构域分子 微小RNA-1237-5p 细胞凋亡相关酪氨酸激酶 乳 腺癌 乳腺癌细胞密歇根癌症基金会7号细胞 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]