| Yan Deng,Zhengguang He,Xiaobin Luo,Rong Qiu,Yong Zhao,Wen Luo. MiR-183-5p promotes the progression of non-small cell lung cancer through targeted regulation of FOXO1. Oncol Transl Med, 2023, 9: 121-132. |
| MiR-183-5p通过靶向调控FOXO1促进非小细胞肺癌的进展 |
| MiR-183-5p promotes the progression of non-small cell lung cancer through targeted regulation of FOXO1 |
| Received:October 12, 2022 Revised:May 23, 2023 |
| DOI：10.1007/s10330-022-0601-1 |
| 中文关键词: 非小细胞肺癌、miR-183-5p、叉头盒蛋白O1、恶性生物学行为；靶向调控 |
| 英文关键词: non-small cell lung cancer; miR-183-5p; forkhead box protein O1; malignant biological behavior; targeted regulation |
| 基金项目: |
| Author Name | Affiliation | E-mail | | Yan Deng | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | zipydw@163.com | | Zhengguang He | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | | | Xiaobin Luo | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | | | Rong Qiu* | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | | | Yong Zhao | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | | | Wen Luo | Department of Respiratory and Critical Care Medicine, Suining Central Hospital, Suining 629000, China | |
|
| Hits: 4554 |
| Download times: 7314 |
| 中文摘要: |
| 目的探讨miR-183-5p与叉头盒蛋白O1 (FOXO1)的靶向关系及其对非小细胞肺癌(NSCLC)细胞增殖、迁移、侵袭和上皮间质转化(EMT)的影响。方法60例非小细胞肺癌患者经病理活检或术中切除获得非小细胞肺癌组织和癌旁正常组织。体外培养的细胞系包括人正常肺上皮细胞系BEAS-2B和人NSCLC细胞系A549、SPCA-1、PC-9和95Mel d。采用qRT-PCR检测组织和细胞中miR-183-5p和foxo1mrna的表达，采用Pearson检验分析它们在NSCLC组织中的相关性，并分析miR-183-5p的表达与临床病理参数的关系。生物信息学预测和双荧光素酶实验验证了miR-183-5p对FOXO1的靶向调控。将A549细胞分为对照组、anti-miR-NC组、anti-miR-183-5p组、miR-NC组、miR-183-5p组、miR-183-5p+pcDNA3.1组和miR-183-5p+pcDNA3.1- foxo1组，分别转染相应的质粒，采用MTT法、克隆形成试验、Transwell室试验、划痕试验和流式细胞术检测A549细胞的增殖、侵袭、迁移、凋亡和细胞周期分布。Western blot检测emt相关蛋白在细胞中的表达。结果miR-183-5p在NSCLC组织和细胞中显著高表达，FOXO1 mRNA表达显著下调。在NSCLC组织中两者呈显著负相关(P < 0.05)。在NSCLC患者中，miR-183-5p表达与肿瘤大小、肿瘤分化及TNM分期显著相关(P < 0.05)。miR-183-5p靶向抑制FOXO1表达与anti-miR-NC组比较，anti-miR-183-5p组OD值、划痕愈合率、E-cadherin、α-catenin蛋白表达量和S期细胞比例显著降低，E-cadherin、α-catenin蛋白表达量和G0/G1期细胞比例显著升高，单克隆形成和侵袭数量显著降低(P < 0.05)。与miR-NC组比较，miR-183-5p组细胞OD值、划痕愈合率、E-cadherin、α-catenin蛋白表达量及S期细胞比例显著升高，E-cadherin、α-catenin蛋白表达量及G0/G1期细胞比例显著降低，单克隆形成数及侵袭数显著升高(P < 0.05)。与miR-183-5p+pcDNA3.1- foxo1组比较，miR-183-5p+pcDNA3.1- foxo1组OD值、划痕愈合率、E-cadherin、α-catenin蛋白表达量及S期细胞比例显著降低，E-cadherin、α-catenin蛋白表达量及G0/G1期细胞比例显著升高。单克隆形成数和侵袭数显著减少(P < 0.05)。结论过表达miR-183-5p可通过下调FOXO1的表达，促进NSCLC细胞的增殖、迁移、侵袭、EMT和细胞周期过程，并抑制细胞凋亡。 |
| 英文摘要: |
| Objective To investigate miR-183-5p targeting to forkhead box protein O1 (FOXO1) and its corresponding
effect on the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of non-small
cell lung cancer (NSCLC) cells.
Methods NSCLC tissues and adjacent normal tissues from 60 patients with NSCLC adenocarcinoma
were obtained via pathological biopsy or intraoperative resection. Several cell lines were cultured in
vitro, including the human normal lung epithelial cell line BEAS-2B and human NSCLC cell lines A549,
SPCA-1, PC-9, and 95-D. miR-183-5p and FOXO1 mRNA expression in tissues and cells were detected
by qRT-PCR; the corresponding correlations in NSCLC tissues were analyzed using the Pearson test,
and the relationship between miR-183-5p expression and clinicopathological parameters was analyzed.
The miR-183-5p-mediated regulation of FOXO1 was verified by bioinformatics prediction alongside double
luciferase, RNA-binding protein immunoprecipitation (RIP) assay, and pull-down experiments. A549 cells
were divided into control, anti-miR-NC, anti-miR-183-5p, miR-NC, miR-183-5p, miR-183-5p+pcDNA3.1,
and miR-183-5p+pcDNA3.1-FOXO1 groups. Cell proliferation, invasion, migration, apoptosis, and cell
cycle distribution were detected using an MTT assay, clone formation assay, Transwell assay, scratch
test, and flow cytometry, respectively. The expression of EMT-related proteins in the cells was analyzed
by western blotting. The effect of miR-185-3p silencing on the development of transplanted tumors was
detected by analyzing tumor formation in nude mice.
Results miR-183-5p expression was significantly higher in NSCLC tissues and cells than in adjacent
normal tissues, whereas FOXO1 mRNA expression was significantly down-regulated. There was a
significant negative correlation between miR-183-5p and FOXO1 mRNA in NSCLC tissues (P < 0.05).
Additionally, the expression of miR-183-5p was significantly correlated with tumor size, tumor differentiation,
and tumor-node-metastasis stage in patients with NSCLC (P < 0.05). miR-183-5p targeted and inhibited
FOXO1 expression. Compared to the anti-miR-NC group, the cell proliferation, scratch healing rate,
N-cadherin and vimentin protein expression, and the proportion of S phase cells were significantly lower in
the anti-miR-183-5p group, whereas the protein expression of E-cadherin and α-catenin and the proportion
of G0/G1 phase cells were significantly higher; additionally, the frequency of colony formation and invasion
were significantly lower in the anti-miR-183-5p group (P < 0.05). Compared to the miR-NC group, the
cell proliferation, scratch healing rate, N-cadherin and vimentin protein expression, and the proportion of
S phase cells in the miR-183-5p group were significantly higher, whereas the E-cadherin and α-catenin
protein expression and the proportion of G0/G1 phase cells were significantly lower; furthermore, the
frequency of colony formation and invasion were significantly higher in the miR-183-5p group (P < 0.05).
Compared with the miR-183-5p+pcDNA3.1 group, the OD value, scratch healing rate, N-cadherin and
vimentin protein expression, and the proportion of S phase cells were significantly lower in the miR-183-
5p+pcDNA3.1-FOXO1 group, whereas E-cadherin and α-catenin protein expression and the proportion of
G0/G1 phase cells were significantly higher; additionally, the frequency of colony formation and invasion
was significantly lower in the miR-183-5p+pcDNA3.1-FOXO1 group (P < 0.05). Overall, silencing miR-185-
3p inhibited the growth of transplanted tumors and promoted FOXO1 expression.
Conclusion Overexpression of miR-183-5p can inhibit apoptosis and promote the proliferation, migration,
invasion, and EMT, of NSCLC cells by down-regulating FOXO1 expression. |
|
View Full Text
Download reader HTML全文 |
| Close |
|
|
|