20 February 2020 : Animal Research
LOC101060264 Silencing Suppresses Invasion and Metastasis of Human Colon Cancer
Weihua Yu1ADE, Yunxia Wang1BCD, Lan Liu1CDE, Shuai Li1BCD, Kongxi Zhu1BDF*DOI: 10.12659/MSM.920270
Med Sci Monit 2020; 26:e920270
Abstract
BACKGROUND: We explored the regulatory effects of long noncoding RNA (lncRNA) LOC101060264 silencing mediated by shRNA on invasion and metastasis of human colon cancer.
MATERIAL AND METHODS: Initially, 2 shRNA plasmids for LOC101060264 silencing – shRNA1 and shRNA2 – were introduced into LoVo cells. Following transfection, the expressions of LOC101060264, E-cadherin, and vimentin were determined. Next, to explore the regulatory effects of LOC101060264 silencing on cell growth, cell cycle, invasion, and migration abilities of LoVo cells, we performed MTT, flow cytometry, Transwell assay, and scratch assay, respectively. Furthermore, in nude mice with xenografted tumors, the tumor volume and weight were measured, and the expressions of PCNA, E-cadherin, vimentin, and MMP-9 in tumor tissues were determined by immunohistochemistry.
RESULTS: The level of E-cadherin increased and the level of vimentin decreased after LOC101060264 silencing mediated by shRNA1 and shRNA2 in LoVo cells. Silencing LOC101060264 repressed the migration, invasion, and proliferation of LoVo cells in vitro and inhibited tumor growth in nude mice in vivo. We also studied the expression of these proteins and found reduced expression of PCNA, vimentin, and MMP-9 protein, and found enhanced expression of E-cadherin protein. Moreover, the inhibitory effect of shRNA2 on the above cell behaviors was stronger than that of shRNA1.
CONCLUSIONS: In summary, LOC101060264 silencing decreased LoVo cell invasiveness via suppressing ETM and attenuated tumor metastasis, which provides a novel therapeutic target for patients with colon cancer.
Keywords: Colorectal Neoplasms, Hereditary Nonpolyposis, RNA Polymerase I, Cadherins, Cell Cycle, Colonic Neoplasms, Down-Regulation, Gene Silencing, Matrix Metalloproteinase 9, Proliferating Cell Nuclear Antigen, Tumor Burden, Vimentin
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