Liu Li-Jie, Yu Jian-Jun, Xu Xiao-Lin
Department of Urology, Shanghai JiaoTong University Affiliated Sixth People's HospitalShanghai 200233, P. R. China.
Department of Urology, Shanghai JiaoTong University Affiliated Sixth People's Hospital South CampusShanghai 201499, P. R. China.
Am J Transl Res. 2017 Jul 15;9(7):3499-3513. eCollection 2017.
We investigated the ability of microRNA-93 (miR-93) to influence proliferation, invasion, migration, and apoptosisofrenal cell carcinoma (RCC) cells via transforming growth factor-β/solvated metal atom dispersed (TGF-β/Smad) signaling by targeting runt-related transcription factor 3 (). RCC tissues with corresponding adjacent normal tissues were collected from 249 RCC patients. And normal renal tissues were collected from patients without RCC who received nephrectomy. The RCC cell line ACHN was treated with miR-93 mimic, mimic-negative control (NC), miR-93 inhibitor, inhibitor-NC, and miR-93 inhibitor + small interfering RNA (siRNA) against (si-RUNX3). Expression of miR-93, RUNX3, TGF-β, and Smad4 were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Cell proliferation was assessed by the Metallothioneins (MTS) assay, cell invasion by the wound-healing assay, cell migration by the Transwell assay, and cell cycle and apoptosis by flow cytometry. Compared with normal renal tissues, the expression of miR-93 and TGF-β were higher while that of RUNX3 and Smad4 were low in RCC and adjacent normal tissues (all <0.05). was confirmed as a target of miR-93 by the dual luciferase reporter gene assay. Compared with mimic-NC group, cell proliferation, invasion, migration and cells from G0/G1 to S phase enhanced but the apoptosis decreased in the miR-93 mimic group (all <0.05). Compared with inhibitor-NC group, proliferation, invasion, and migration reduced, while apoptosis increased, and cells at G0/G1 phase arrested in the miR-93 inhibitor group (all <0.05). Compared with miR-93 inhibitor group, cell proliferation, invasion, and migration increased with increasing cells from G1 to S phase while the apoptosis decreased, in miR-93 inhibitor + si-RUNX3 group (all <0.05). In conclusion, miR-93 inhibits apoptosis and promotes proliferation, invasion, and migration of RCC cells via TGF-β/Smad signaling by inhibiting .
我们研究了微小RNA-93（miR-93）通过靶向 runt相关转录因子3（RUNX3），经由转化生长因子-β/溶剂化金属原子分散体（TGF-β/Smad）信号通路影响肾细胞癌（RCC）细胞增殖、侵袭、迁移和凋亡的能力。收集了249例RCC患者的RCC组织及其相应的癌旁正常组织。并从接受肾切除术的非RCC患者中收集正常肾组织。用miR-93模拟物、模拟物阴性对照（NC）、miR-93抑制剂、抑制剂-NC以及针对RUNX3的miR-93抑制剂+小干扰RNA（siRNA）（si-RUNX3）处理RCC细胞系ACHN。通过定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法评估miR-93、RUNX3、TGF-β和Smad4的表达。通过金属硫蛋白（MTS）试验评估细胞增殖，通过伤口愈合试验评估细胞侵袭，通过Transwell试验评估细胞迁移，通过流式细胞术评估细胞周期和凋亡。与正常肾组织相比，RCC组织及癌旁正常组织中miR-93和TGF-β的表达较高，而RUNX3和Smad4的表达较低（均P<0.05）。双荧光素酶报告基因试验证实RUNX3是miR-93的靶标。与模拟物-NC组相比，miR-93模拟物组细胞增殖、侵袭、迁移以及从G0/G1期到S期的细胞增多，但凋亡减少（均P<0.05）。与抑制剂-NC组相比，miR-93抑制剂组细胞增殖、侵袭和迁移减少，而凋亡增加，且细胞停滞于G0/G1期（均P<0.05）。与miR-93抑制剂组相比，miR-93抑制剂+si-RUNX3组细胞增殖、侵袭和迁移增加，从G1期到S期的细胞增多，而凋亡减少（均P<0.05）。总之，miR-93通过抑制RUNX3，经由TGF-β/Smad信号通路抑制RCC细胞凋亡并促进其增殖、侵袭和迁移。
