LIM kinase 1 serves an important role in the multidrug resistance of osteosarcoma cells

Yang,Jian‑Zeng; Huang,Li‑Hong; Chen,Rui; Meng,Ling‑Jie; Gao,Yi‑Yao; Ji,Qiu‑Ye; Wang,Yan

doi:10.3892/ol.2017.7317

LIM kinase 1 serves an important role in the multidrug resistance of osteosarcoma cells

Authors:
- Jian‑Zeng Yang
- Li‑Hong Huang
- Rui Chen
- Ling‑Jie Meng
- Yi‑Yao Gao
- Qiu‑Ye Ji
- Yan Wang
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Affiliations: Scientific Research Center, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China, Department of Physiology and Medicine, China‑Japan Union Hospital of Jilin University, Changchun, Jilin 130033, P.R. China
Published online on: November 1, 2017 https://doi.org/10.3892/ol.2017.7317
Pages: 250-256
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Multidrug resistance (MDR) is a major challenge for the management of the majority of cancers. The precise molecular mechanisms of MDR remain elusive. In a previous study, a multidrug resistant osteosarcoma model [MG63/vincristine (VCR)] was established by intermittent exposure of MG63 cells to gradually increasing concentrations of VCR. These cells exhibited cross‑resistance to multiple structurally and mechanistically unrelated chemotherapeutic agents. The development of MDR was associated with increased expression of LIM kinase 1 (LIMK1). Compared with that in normal human fetal osteoblasts (hFOB) 1.19, the messenger RNA and protein expression of LIMK1 was signiﬁcantly elevated both in MG63 and U2OS osteosarcoma cells. To observe the expression pattern of LIMK1 in osteosarcoma, immunohistochemical analyses were performed on specimens derived from 6 patients. The results indicated that LIMK1 was expressed to a greater extent in the tumor parenchyma than in the mesenchyme. The role of LIMK1 in MDR was confirmed by transfecting plasmids coding LIMK1‑small interfering RNA (siRNA), wild‑type‑LIMK1 or empty vector into MG63/VCR cells, and measuring the expression of LIMK1 and multidrug resistance protein 1 (MDR1), also known as P‑gycoprotein (P‑gp). The results demonstrated that the level of MDR1/P‑gp was positively correlated with the level of LIMK1. This correlation was also shown with the doxorubicin efflux assay and by measuring apoptosis. Specifically, after 6 h of incubation with VCR, 25.6% of the cells transfected with the LIMK1‑siRNA plasmid were apoptotic compared with 6.2% in the empty vector group and 1.3% in the group of cells transfected with the wild‑type‑LIMK1 plasmid. Thus, it was concluded that LIMK1 serves a key role in the MDR of osteosarcoma and functions through MDR1.

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