MiR-1246 promotes SiHa cervical cancer cell proliferation, invasion, and migration through suppression of its target gene thrombospondin 2
MiR-1246 promotes SiHa cervical cancer cell proliferation, invasion, and migration through suppression of its target gene thrombospondin 2
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MiR-1246通过抑制靶基因血小板反应蛋白2促进SiHa宫颈癌细胞增殖、侵袭和迁移
DOI：
10.1007/s00404-014-3260-2
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发表时间：
2014-10-01
影响因子：
2.6
通讯作者：
Long, Fengyi
中科院分区：
文献类型：
--
作者：
Chen, Junying;Yao, Desheng;Long, Fengyi
探讨miR-1246对人（CSCC）细胞株SiHa增殖、侵袭和迁移的影响。SiHa细胞分为三组：miR-1246类似物；mir - 1246拮抗剂;和控制。MTT、transwell和伤口愈合试验分别评估SiHa细胞的增殖、侵袭和迁移能力。Western blot检测miR-1246类似物、拮抗剂或对照转染前后血栓反应蛋白-2 （THBS2）的蛋白表达。此外，生成含有THBS2 3'- utr的双荧光素酶质粒，并与抑制剂miR-1246或非特异性miRNA共转染SiHa细胞，观察其对THBS2驱动的荧光素酶活性的影响。MTT、transwell和伤口愈合实验显示，转染miR-1246类似物的SiHa细胞的增殖、迁移和侵袭均显著增强（P < 0.01），而转染miR-1246拮抗剂的SiHa细胞的增殖、迁移和侵袭均受到抑制。Western blot数据显示，miR-1246类似物转染的SiHa细胞与对照转染的细胞相比，THBS2表达显著降低（灰色值= 6.28 +/- A 10.22 vs. 9.58 +/- A 17.58; P = 0.013），而转染miR-1246拮抗剂的SiHa细胞THBS2表达显著增加（灰色值= 12.90 +/- A 19.81; P = 0.037）。此外，与对照组相比，共转染miR-1246和含THBS2 3'- utr质粒的SiHa细胞表现出较低的荧光素酶活性。MiR-1246诱导CSCC SiHa细胞增殖、侵袭和迁移。初步证据表明，miR-1246可能通过抑制其靶基因THBS2促进CSCC的发生和进展。
To investigate the effects of miR-1246 on proliferation, invasion, and migration in the human (CSCC) cell line SiHa.SiHa cells were assigned into three groups: miR-1246 analog; miR-1246 antagonist; and control. The MTT, transwell, and wound healing assays were performed to evaluate the proliferation, invasion, and migration abilities of SiHa cells, respectively. Western blot was carried out to detect protein expression of thrombospondin-2 (THBS2) before and after transfection with miR-1246 analog, antagonist, or control. In addition, a THBS2 3'-UTR-containing dual luciferase plasmid was generated and co-transfected with miR-1246, the inhibitor, or non-specific miRNA, into SiHa cells to observe its effects on THBS2-driven luciferase enzyme activity.MTT, transwell, and wound healing assays revealed that proliferation, migration, and invasion were all significantly enhanced (P < 0.01) in SiHa cells transfected with miR-1246 analog, but were suppressed in those transfected with the miR-1246 antagonist. Western blot data showed that miR-1246 analog-transfected SiHa cells had significantly decreased THBS2 expression when compared with control-transfected cells (gray value = 6.28 +/- A 10.22 vs. 9.58 +/- A 17.58; P = 0.013) while those transfected with the miR-1246 antagonist had significantly increased THBS2 expression (gray value = 12.90 +/- A 19.81; P = 0.037). Moreover, SiHa cells co-transfected with miR-1246 and the THBS2 3'-UTR-containing plasmid exhibited decreased luciferase enzyme activity compared with the control.MiR-1246 induced CSCC SiHa cell proliferation, invasion and migration. Preliminary evidence suggests that miR-1246 might promote CSCC tumorigenesis and progression by the suppression of its target gene THBS2.