miR-135a inhibits malignant proliferation and diffusion of non-small cell lung cancer cells by down-regulating ROCK1 protein

Abstract

Objective: To seek the clinical significance and regulatory mechanism of miR-135a and Rho-associated protein kinase 1 (ROCK1) in non-small cell lung cancer (NSCLC). Methods: NSCLC cells were purchased, and miR-135a-mimics, miR-135a-inhibitor, miR-NC, si-ROCK1 and Sh-ROCK1 were transfected into NSCLC cells HCC827 and NCI-H524. qRT-PCR and Western blot were used to detect the expression of miR-135a, ROCK1, Bax, Caspase3, Bcl-2, N-cadherin, vimentin and E-cadherin. MTT, scratch test, Transwell and flow cytometry were used to analyze the cell proliferation, migration, invasion and apoptosis. Results: miR-135a was low expressed in serum of NSCLC group, while ROCK1 was opposite. miR-135a low level or ROCK1 high level was associated with poor prognosis of NSCLC and lower 3-year OS. Over-expression of miR-135a and inhibition of ROCK1 expression could control malignant growth and diffusion of cells and expression of Bcl-2, N-cadherin and vimentin proteins, and promote apoptosis and expression of Bax, Caspase3 and E-cadherin proteins. After transfection of miR-135a-mimics+sh-ROCK1 to HCC827 and NCI-H524, the malignant proliferation and diffusion behavior of the cells were not different from those of the miR-NC group with no transfection sequence. The double luciferase report revealed that miR-135a has a targeting relationship with ROCK1. Conclusion: miR-135a is abnormally down-regulated in NSCLC. As a serum indicator, miR-135a has the potential to diagnose NSCLC and predict prognosis. The up-regulated expression of miR-135a protein can down-regulate the ROCK1 protein, inhibit the malignant proliferation, migration, invasion, EMT and other diffusion behaviors of NSCLC cells, and increase the apoptosis ability of cells.

Circ‐N4BP2L2 enhances mitochondrial function in non‐small cell lung cancer cells through regulating the miR‐135a‐5p/ARL5B axis

“…In this study, miR‐135a‐5p was demonstrated to be downregulated in NSCLC tissues and cells, which was consistent with a previous study 18 . Low expression of miR‐135a was associated with poor prognosis of NSCLC patients, and miR‐135a had the potential to be a serum indicator for diagnosing NSCLC 19 . MiR‐135a‐5p could be sponged by circLARP4 in NSCLC 20 .…”

“…18 Low expression of miR-135a was associated with poor prognosis of NSCLC patients, and miR-135a had the potential to be a serum indicator for diagnosing NSCLC. 19 MiR-135a-5p could be sponged by circLARP4 in NSCLC. 20 Moreover, we found that ARL5B was a target gene of miR-135a-5p, and was negatively regulated by miR-135a-5p.…”

Implication of non-coding RNA-mediated ROCK1 regulation in various diseases

“…A number of ROCK1-interacting miRNAs have been found to reduce tumor burden. For instance, experiments in non-small cell lung carcinoma cells showed that the tumor suppressor roles of miR-135a ( Zhao et al, 2020 ), miR-148b ( Luo and Liang, 2018 ) and miR-335-5p ( Du et al, 2019 ) are exerted through modulation of expression of ROCK1. The interactions between miRNAs and ROCK1 have been mostly assessed in osteosarcoma cells among other cancers.…”