Department of Gastroenterology, The Second Hospital of Shandong University, Jinan, Shandong, China (mainland).
Med Sci Monit. 2020 Feb 20;26:e920270. doi: 10.12659/MSM.920270.
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.
我们探讨了短发夹 RNA(shRNA)介导的长链非编码 RNA(lncRNA)LOC101060264 沉默对人结肠癌细胞侵袭和转移的调控作用。
首先,将 2 个用于 LOC101060264 沉默的 shRNA 质粒(shRNA1 和 shRNA2)导入 LoVo 细胞。转染后,检测 LOC101060264、E-钙黏蛋白和波形蛋白的表达。接下来,为了探讨 LOC101060264 沉默对 LoVo 细胞生长、细胞周期、侵袭和迁移能力的调控作用,分别进行 MTT、流式细胞术、Transwell 检测和划痕实验。此外,在裸鼠皮下移植瘤模型中,测量肿瘤体积和重量,并通过免疫组化检测肿瘤组织中 PCNA、E-钙黏蛋白、波形蛋白和 MMP-9 的表达。
shRNA1 和 shRNA2 介导的 LOC101060264 沉默后,LoVo 细胞中 E-钙黏蛋白表达上调,波形蛋白表达下调。沉默 LOC101060264 抑制 LoVo 细胞在体外的迁移、侵袭和增殖,并抑制裸鼠体内肿瘤的生长。我们还研究了这些蛋白的表达,发现 PCNA、波形蛋白和 MMP-9 蛋白表达降低,E-钙黏蛋白蛋白表达增加。此外,shRNA2 对上述细胞行为的抑制作用强于 shRNA1。
总之,LOC101060264 沉默通过抑制 EMT 降低 LoVo 细胞的侵袭性,并减弱肿瘤转移,为结肠癌患者提供了一个新的治疗靶点。