| 任继兵,陈梦琳,李传斌.长链基因间非编码 RNA 00963通过靶向微小 RNA-148b-3p对结直肠癌细胞增殖、迁移和侵袭的影响[J].安徽医药,2021,25(10):2093-2098. |
| 长链基因间非编码 RNA 00963通过靶向微小 RNA-148b-3p对结直肠癌细胞增殖、迁移和侵袭的影响 |
| Effect of long intergenic non-coding RNA 00963 (LINC00963) on the proliferation, migration and invasion of colorectal cancer cells by targeting microRNA-148b-3p (miR-148b-3p) |
| |
| DOI：10.3969/j.issn.1009-6469.2021.10.043 |
| 中文关键词: 结直肠肿瘤 长链基因间非编码 RNA00963 微小 RNA-148b-3p 增殖 迁移 侵袭 |
| 英文关键词: Colorectal neoplasms LINC00963 miR-148b-3p Proliferation Migration Invasion |
| 基金项目: |
|
| 摘要点击次数: 1242 |
| 全文下载次数: 447 |
| 中文摘要: |
| 目的探讨长链基因间非编码 RNA00963(LINC00963)对结直肠癌细胞恶性生物学行为的影响及作用机制。方法本研究于 2018年 8月至 2019年 5月进行；正常结直黏膜上皮细胞 NCM460以及结直肠癌细胞系 HT29、SW480、SW1116购自中国科学院上海细胞库，实时荧光定量 PCR(qRT-PCR)检测微小 RNA-148b-3p(miR-148b-3p)和 LINC00963的表达水平；双荧光素酶报告实验检测 miR-148b-3p和 LINC00963的靶向关系。将 HT29细胞分为 miR-NC组、 miR-148b-3p组、 si-NC组、 siLINC00963组、 si-LINC00963+anti-miR-NC组、 si-LINC00963+anti-miR-148b-3p组；蛋白质印迹法( Western Blotting)检测蛋白表达；四甲基偶氮唑盐比色法( MTT)检测细胞活性； Transwell检测各组细胞迁移和侵袭。结果与 NCM460细胞相比， HT29、 SW480、SW1116细胞中 miR-148b-3p的表达水平显著降低［( 0.32±0.04)、(0.48±0.04)、(0.39±0.04)比( 1.03±0.09)］，LINC00963的表达水平显著升高［(3.13±0.31)、(2.76±0.27)、(2.91±0.28)比( 1.00±0.09)］差异有统计学意义(P<0.05)。 LINC00963靶向调控 miR-148b-3p的表达。干扰 LINC00963后，细胞周期蛋白 D1(Cyclin D1)、，基质金属蛋白酶 2(MMP-2)、基质金属蛋白酶 9(MMP-9)蛋白表达水平降低，细胞周期蛋白依赖性激酶抑制剂 1A(p21)表达水平升高， HT29细胞活性［48 h为( 0.43±0.04)比(0.65±0.06)， 72 h为( 0.56±0.05)比( 1.05±0.09)］降低，及迁移数量减少［( 39.65±3.42)个比( 81.24±8.06)个］侵袭数量减少［( 32.14±3.28)个比( 69.54±6.27)个］差异有统计学意义( P<0.05)。过表达 miR-148b-3p后， Cyclin D1、MMP-2、M，MP-9表达水平降低， p21表达水平升高， HT29细胞活，性［48 h为( 0.49±0.04)比( 0.67±0.06)72 h为( 0.62±0.06)比( 1.06±0.09)］降低，及迁移数量减少［( 45.32±4.28)个比( 86.27±8.25)个］，侵袭数量减少［( 39.54±4.03)个，比( 73.65±7.24)个］，差异有统计学意义( P<0.05)。抑制 miR-148b-3p表达可逆转干扰 LINC00963对 HT29细胞的作用。结论干扰 LINC00963可通过上调 miR-148b-3p抑制结直肠癌 HT29细胞的增殖、迁移和侵袭。 |
| 英文摘要: |
| Objective To explore the effect and mechanism of long-chain intergenic non-coding RNA 00963 (LINC00963) on the malignant biological behavior of colorectal cancer cells.Methods This experiment was conducted from August 2018 to May 2019, normal colorectal mucosal epithelial cells NCM460 and colorectal cancer cell lines HT29, SW480, SW1116 were purchased from theShanghai Cell Bank of the Chinese Academy of Sciences, real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression levels of microRNA-148b-3p (miR-148b-3p) and LINC00963; double luciferase reporter experiment was used to detect the targeting relationship between miR-148b-3p and LINC00963. HT29 cells were assigned into miR-NC group, miR-148b-3p group, si-NC group, si-LINC00963 group, si-LINC00963+anti-miR-NC group, si-LINC00963+anti-miR-148b-3p group; Western blotting was used todetect protein expression; tetramethylazolium salt colorimetric method (MTT) to detect cell viability; Transwell was used to detect cellmigration and invasion in each group.Results Compared with NCM460 cells, the expression level of miR-148b-3p in HT29, SW480 and SW1116 cells was significantly reduced [(0.32±0.04), (0.48±0.04), (0.39±0.04) vs. (1.03±0.09)], LINC00963 expression level was significantly increased [(3.13±0.31), (2.76±0.27), (2.91±0.28) vs. (1.00±0.09)], the difference was statistically significant (P<0.05). LINC00963 targeted the expression of miR-148b-3p. After interference with LINC00963, Cyclin D1 , matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 9 (MMP-9) protein expression levels were decreased, and cyclin-dependent kinase inhibitor 1A (p21) expression level was increased, the activity of HT29 cells [48 h: (0.43±0.04) vs. (0.65±0.06), 72 h: (0.56±0.05) vs. (1.05±0.09)] was decreased, and the number of migration [(39.65±3.42) vs. (81.24±8.06)] was decreased, the number of invasions was decreased [(32.14± 3.28) vs. (69.54±6.27)], the difference was statistically significant (P<0.05). After over-expression of miR-148b-3p, Cyclin D1, MMP-2, MMP-9 expression levels were decreased, p21 expression was increased, HT29 cell viability [48 h: (0.49±0.04) vs. (0.67±0.06), 72 h : (0.62±0.06) vs. (1.06±0.09)] was decreased, and the number of migration was decreased [(45.32±4.28) vs. (86.27±8.25)], and the number of invasion was decreased [(39.54±4.03) vs. (73.65±7.24)], the difference was statistically significant (P<0.05). Suppressing the ex |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
