Department of Gastrointestinal Surgery, Changshu No. 2 Hospital , Suzhou, Jiangsu, China.
Department of General Surgery, The First Affiliated Hospital of Soochow University , Suzhou, Jiangsu, China.
Bioengineered. 2021 Dec;12(1):627-639. doi: 10.1080/21655979.2021.1882164.
We aimed to study the role of LINC00997 in the metastasis of colorectal cancer (CRC). LINC00997 and miR-512-3p expression in the primary colorectal cancer (NCRC) tissues and metastatic colorectal cancer (MCRC) tissues were detected using RT-qPCR. The Cancer Genome Atlas database was used to evaluate LINC00997 levels in the NCRC and MCRC tissues, and the correlations of LINC00997 expression with distant metastasis (M), regional lymph node metastasis (N), age and tumor stage were analyzed. Subsequently, RT-qPCR was performed to determine the expression of metastasis-related genes in MCRC tissues and analyze the correlation of LINC00997 or miR-512-3p level with the protein expression of metastasis-related genes. , LINC00997 expression in several CRC cell lines was examined. After LINC00997 silencing, cell invasion and migration were evaluated with Transwell and wound healing assays, respectively. The expression of metastasis- and EMT-related proteins was measured. Additionally, the potential interaction between LINC00997 and miR-512-3p was verified using a luciferase reporter assay. Rescue assays were conducted to clarify the regulatory effects of LINC00997 and miR-512-3p on CRC development. Results revealed that LINC00997 was frequently overexpressed in MCRC tissues, which was positively related to the tumor metastasis and stage. Additionally, LINC00997 was significantly elevated in CRC cells and LINC00997 silencing inhibited the invasion, migration and EMT of CRC cells, which was restored by miR-512-3p inhibitor. Luciferase reporter assay confirmed that LINC00997 could target miR-512-3p. In conclusion, LINC00997 regulated the metastasis of CRC by targeting miR-512-3p, providing some insights into the regulatory mechanism of CRC.
我们旨在研究 LINC00997 在结直肠癌(CRC)转移中的作用。使用 RT-qPCR 检测原发性结直肠癌(NCRC)组织和转移性结直肠癌(MCRC)组织中的 LINC00997 和 miR-512-3p 的表达。使用癌症基因组图谱数据库评估 NCRC 和 MCRC 组织中的 LINC00997 水平,并分析 LINC00997 表达与远处转移(M)、区域淋巴结转移(N)、年龄和肿瘤分期的相关性。随后,通过 RT-qPCR 测定 MCRC 组织中转录相关基因的表达,并分析 LINC00997 或 miR-512-3p 水平与转移相关基因蛋白表达的相关性。还检测了几种 CRC 细胞系中的 LINC00997 表达。沉默 LINC00997 后,分别用 Transwell 和划痕愈合实验评估细胞侵袭和迁移能力。测量转移和 EMT 相关蛋白的表达。此外,通过荧光素酶报告基因实验验证 LINC00997 和 miR-512-3p 之间的潜在相互作用。进行挽救实验以阐明 LINC00997 和 miR-512-3p 对 CRC 发展的调节作用。结果表明,LINC00997 在 MCRC 组织中频繁过表达,与肿瘤转移和分期呈正相关。此外,LINC00997 在 CRC 细胞中显著上调,LINC00997 沉默抑制 CRC 细胞的侵袭、迁移和 EMT,这可被 miR-512-3p 抑制剂所恢复。荧光素酶报告基因实验证实 LINC00997 可以靶向 miR-512-3p。总之,LINC00997 通过靶向 miR-512-3p 调节 CRC 的转移,为 CRC 的调控机制提供了一些见解。