MicroRNA-93 inhibits apoptosis and promotes proliferation, invasion and migration of renal cell carcinoma ACHN cells via the TGF-β/Smad signaling pathway by targeting
Authors: Li-Jie Liu
Jian-Jun Yu
Xiao-Lin Xu
Journal: American journal of translational research (2017)
Discipline: BIOLOGY & BIOCHEMISTRY
Keywords: Smad4; invasion; microRNA-93; migration; proliferation
Affiliations: 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.

Abstract:  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 (RUNX3). 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 RUNX3 (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 P&lt;0.05). RUNX3 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 P&lt;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 P&lt;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 P&lt;0.05). In conclusion, miR-93 inhibits apoptosis and promotes proliferation, invasion, and migration of RCC cells via TGF-β/Smad signaling by inhibiting RUNX3. 

Retracted: No

PubPeer comments:
 Fig 7B.  Effect of miR-93 on invasion of RCC cells using Transwell assay . ![file] After inversion, one panel overlaps with a panel of Fig 5A B from [Lu et al (2018)] (orange rectangles). ![file] Two panels have corresponding version in Fig 7B from [Wu et al (2017)] (pink, light blue rectangles). ![file] , 
 *[right]* Fig 2C D.  Grey value of target proteins of Smad4; ... Grey value of target proteins of TGF-u03b2 . *[left]* Fig 7f h from [Zhang et al (2018)] ,  The protein expression of PCNA and Ki67 in ... HepG2 cells of each group.... The protein expression of cyclins (cyclin E, CDK2 and p21) in ... HepG2 cells of each group. ![file] These are very versatile blots. ---------------------------------------------------- The Figure legend goes on to explain that  Smad, solvated metal atom dispersed . Really? , 
 *[right]* Fig 4D,  The protein expression of RUNX3 in each groups using western blotting . *[Left]* Fig 1A from [Xia et al (2018)] ,  protein bands of NFkB expression among 5 groups . ![file] Surprisingly similar loading controls. ,