miR-302b-3p靶向SQSTM1调控乳腺癌MCF7细胞的自噬  

miR-302b-3p targets SQSTM1 to regulate autophagy in breast cancer cell MCF7

作　　者：陈晶[1] 白勇 木拉提.库尔班 孙国辉[1] 叶斯波力 CHEN Jing;BAI Yong;Mulati Kulban;SUN Guohui;Yesperi(Department of Oncology,The Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830063,China)

机构地区：[1]新疆医科大学第二附属医院肿瘤科,新疆乌鲁木齐830063

出　　处：《解剖学研究》2019年第2期110-114,共5页Anatomy Research

摘　　要：目的探讨miR-302b-3p靶向SQSTM1调控乳腺癌MCF7细胞的自噬。方法运用TargetScan在线分析miR-302b-3p与SQSTM1的相关性;将SQSTM1的3′UTR构建进PMIR-RB-REPORT质粒,利用luciferase assay检测miR-302b-3p是否靶向调控SQSTM1;用RNA转染试剂转染miR-302b-3p mimics或miR-302b-3p inhibitor进乳腺癌MCF7细胞,通过Western blot检测SQSTM1的表达量和乳腺癌MCF7细胞自噬标志蛋白表达情况。单丹磺酰尸胺染色检测细胞自噬发生水平。结果 miR-302b-3p靶向SQSTM1的3′UTR的584-590的位置;过表达miR-302b-3p时,SQSTM1的表达量明显下降(P<0.05),细胞自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ和Beclin1的表达量明显上升(P<0.05),MCF7自噬发生水平降低(P<0.05);敲低miR-302b-3p时,SQSTM1的表达量明显上升(P<0.05),细胞自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ和Beclin1的表达量明显下降(P<0.05),MCF7自噬发生水平升高(P<0.05)。结论 miR-302b-3p靶向SQSTM1的3′UTR后降低SQSTM1的蛋白水平,最终抑制乳腺癌细胞MCF7的自噬。Objective To investigate whether miR-302b-3p targets SQSTM1 to regulate autophagy in breast cancer cells MCF7. Methods TargetScan was used to analyze the correlation between miR-302b-3p and SQSTM1;the 3′UTR of SQSTM1 was constructed into PMIR-RB-REPORT plasmid,and luciferase assay was used to detect whether miR-302 b-3 p targets SQSTM1;The staining reagent was transfected into miR-302b-3p mimics or miR-302 b-3p inhibitor into breast cancer MCF7 cells. The expression of SQSTM1 and the expression of autophagy marker protein in breast cancer MCF7 cells were detected by western blot. Monodansylcadaverine staining was used to detect the level of autophagy. Results miR-302b-3p targeted the position of 584-590 of the 3′UTR of SQSTM1;when overexpressing miR-302b-3p,the expression of SQSTM1 was significantly decreased(P<0.05),autophagy-associated protein LC3-Ⅱ The expression of/LC3-Ⅰ and Beclin1 was significantly increased(P<0.05),and the level of autophagy was decreased in MCF7(P<0.05). The expression of SQSTM1 was significantly increased when knocking down miR-302b-3p(P<0.05). The expression levels of autophagy-related proteins LC3-Ⅱ/LC3-Ⅰ and Beclin1 were significantly decreased(P<0.05),and the levels of autophagy in MCF7 were increased(P<0.05). Conclusion miR-302b-3p targets the 3′UTR of SQSTM1 and decreases the protein level of SQSTM1,which ultimately inhibits autophagy in breast cancer cells MCF7.

关 键 词：乳腺癌 MCF7细胞株 miR-302b-3p SQSTM1 自噬 

分 类 号：R737.9[医药卫生—肿瘤]