| 刘锐,褚伟伟,余明军,王海明.LncRNA OIP5-AS1通过海绵吸附miR-143-3p调控COL1A1促进胃癌发展[J].浙江中西医结合杂志,2022,32(9): |
| LncRNA OIP5-AS1通过海绵吸附miR-143-3p调控COL1A1促进胃癌发展 |
| LncRNA OIP5-AS1 regulates COL1A1 to promote the development of gastric cancer through sponges adsorption of miR-143-3p |
| 投稿时间：2022-03-21 修订日期：2022-08-23 |
| DOI： |
| 中文关键词: 胃癌 增殖 迁移 侵袭 lncRNA OIP5-AS1 miR-143-3p COL1A1 |
| 英文关键词:Gastric cancer Proliferation Migration Invasion LncRNA OIP5-AS1 miR-143-3p COL1A1 |
| 基金项目:杭州市科技计划引导项目(No.20201231Y027) |
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| 中文摘要: |
| 摘要 目的 探究长链非编码RNA（LncRNA）Opa相互作用蛋白5反义RNA 1 （OIP5-AS1）对胃癌发展的调控机制。方法 qRT-PCR检测LncRNA OIP5-AS1 在正常胃细胞系（GES-1）和两种胃癌细胞系（AGS和SGC7901）中的表达水平；构建OIP5-AS1敲低载体、miR-143-3p 抑制剂（miR-143-3p inhibitor）以及Ⅰ型胶原α1（COL1A1）敲低载体并转染胃癌细胞系，运用CCK-8实验、伤口愈合实验、Transwell 实验检测 LncRNA OIP5-AS1、miR-143-3p、COL1A1在胃癌细胞增殖、迁移、侵袭中的作用；采用双荧光素酶报告基因、qRT-PCR、Western blot实验检测LncRNA OIP5-AS1以及miR-143-3p、COL1A1之间的靶向调控作用。结果 OIP5-AS1在胃癌细胞AGS、SGC7901中表达水平分别是GES-1细胞的6.10和5.53倍，差异有统计学意义(P<0.01)。同时，敲低 OIP5-AS1会抑制胃癌细胞增殖[AGS：（0.71±0.07）比（1.20±0.11）；SGC7901：（1.02±0.10）比（1.80±0.10），P均<0.05]、迁移[AGS：（29.01±3.68）%比(72.04±6.82)%；SGC7901：（23.97±2.24）%比(62.02±3.73)%，P均<0.01]和侵袭[AGS：（35.68±6.96）比(278.33±11.85)；SGC7901： （62.74±12.77）比(248.65±12.03)，P均<0.01]。双荧光素酶报告基因证实OIP5-AS1靶向作用 miR-143-3p 并下调其表达水平，miR-143-3p 可负调控 COL1A1 的表达(P<0.01)。进一步研究发现，敲降 OIP5-AS1通过靶向上调miR-143-3p对COL1A1的抑制作用，抑制胃癌细胞增殖[AGS：(0.56±0.06)比(1.29±0.08)；SGC7901：(0.85±0.05) 比(1.94±0.10)，P均<0.01]、迁移[AGS：(26.33±4.19) %比 (69.67±2.87) %；SGC7901：(23.00±2.94) %比(63.33±3.30) % ，P均<0.01]和侵袭[AGS：(34.00±7.48)比(272.33±14.88) ；SGC7901： (55.33±9.53)比(274.67±8.96)，P均<0.01]。结论 lncRNA OIP5-AS1通过海绵吸附miR-143-3p进而上调COL1A1促进胃癌细胞增殖、迁移和侵袭，为胃癌临床诊断与治疗提供了潜在的标靶。 |
| 英文摘要: |
| ABSTRACT Objective To investigate the regulatory mechanism of long non-coding RNA (LncRNA) Opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) on the development of gastric cancer. Methods qRT-PCR was used to detect the expression levels of LncRNA OIP5 - AS1 in normal gastric cell line (GES-1) and two gastric cancer cell lines (AGS and SGC7901). The OIP5-AS1 knock-down vector, miR-143-3p inhibitor and COL1A1 knock-down vector were constructed and transfected into gastric cancer cell lines. The CCK-8, wound healing and Transwell experiment were used to detect the effects of LncRNA OIP5 - AS1, miR-143-3p and COL1A1 on the proliferation, migration and invasion of gastric cancer cells. The Dual-luciferase reporter genes, qRT-PCR and Western blot experiments were used to detect the targeting regulation between LncRNA OIP5-AS1, miR-143-3p and COL1A1. Results The expression level of OIP5-AS1 in gastric cancer cells AGS and SGC7901 was 6.10 and 5.53 times higher than that in GES-1 cells, respectively (P<0.01).At the same time, Knockdown of OIP5-AS1 inhibited gastric cancer cell proliferation [AGS,（0.71±0.07）vs.（1.20±0.11）;SGC7901,（1.02±0.10）vs.（1.80±0.10），all P<0.05], migration [AGS,( 29.01±3.68 )% vs.（72.04±6.82）%; SGC7901,( 23.97±2.24)% vs.（62.02±3.73）%，all P<0.01] and invasion [AGS,( 35.68±6.96) vs.（278.33±11.85）；SGC7901,( 62.74±12.77) vs.（248.65±12.03）,all P<0.01].The dual-luciferase reporter confirmed that OIP5-AS1 targeted miR-143-3p and downregulated its expression level, and miR-143-3p could negatively regulate COL1A1 expression (P< 0.01).Further studies revealed that the knockdown of OIP5-AS1 inhibited COL1A1 through a targeted upregulation of miR-143-3p, Inhibition of gastric cancer cell proliferation [AGS,(0.56±0.06) vs. (1.29±0.08)；SGC7901,(0.85±0.05) vs. (1.94±0.10)，all P<0.01], and migration [AGS，(26.33±4.19) % vs. (69.67±2.87) %；SGC7901，(23.00±2.94) % vs. (63.33±3.30) %，all P <0.01] and the invasion [AGS，(34.00±7.48) vs. (272.33±14.88)；SGC7901，(55.33±9.53) vs. (274.67±8.96)，all P <0.01].Conclusion The lncRNA OIP5-AS1 upregulates COL1A1 through sponge adsorption of miR-143-3p to promote the proliferation, migration and invasion of gastric cancer cells, providing a potential target for clinical diagnosis and treatment of gastric cancer. |
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