目的: 探讨miR-17-5p在肺腺癌细胞中的表达及miR-17-5p对肺腺癌细胞增殖、侵袭、迁移、凋亡的影响及其分子作用机制。方法: 采用实时荧光定量PCR检测人肺腺癌细胞系HCC827和人正常肺上皮细胞BEAS-2B中miR-17-5p的表达水平。实验分为miR-17-5p过表达组(miR-17-5p mimics组)、miR-17-5p低表达组(miR-17-5p inhibitor组)和阴性对照组(NC组)。通过CCK-8、流式细胞术、Transwell实验探讨miR-17-5p对肺腺癌细胞增殖、凋亡、侵袭及迁移的影响。最后通过Targetscan数据库预测出PKD2是miR-17-5p下游潜在的靶基因,利用实时荧光定量PCR和Western-blot实验验证miR-17-5p与PKD2之间的相互作用关系。结果: miR-17-5p在HCC827细胞中高表达(P＜0.001),上调miR-17-5p后显著促进了肺腺癌细胞增殖(P＜0.05)、侵袭(P＜0.01)及迁移(P＜0.05)并抑制凋亡(P＜0.01)。miR-17-5p与PKD2的3’UTR存在直接结合位点,并且过表达miR-17-5p在mRNA和蛋白水平均显著促进了PKD2的表达(P＜0.01) 。结论: PKD2是miR-17-5p的直接靶基因,miR-17-5p通过调控PKD2影响肺腺癌细胞的生物学行为。
Objective: To investigate the expression of miR-17-5p in lung adenocarcinoma cells, and to study the effects of miR-17-5p on proliferation, invasion, migration and apoptosis of lung adenocarcinoma cells and its molecular mechanism. Methods: The expression of miR-17-5p in human lung adenocarcinoma cell line HCC827 and human normal lung epithelial cell line BEAS-2B were detected by real-time fluorescent quantitative PCR. There were miR-17-5p mimics group, miR-17-5p inhibitor group and NC group in the experiment. The effects of miR-17-5p on proliferation, apoptosis, invasion and migration of lung adenocarcinoma cells were analyzed by CCK-8, flow cytometry and Transwell assay. Finally, PKD2 was predicted to be a potential downstream target gene of miR-17-5p by Targetscan database. The interaction between miR-17-5p and PKD2 was verified by real-time fluorescence quantitative PCR and Western-blot experiments. Results: miR-17-5p was highly expressed in HCC827 cells (P＜0.001), and up-regulation of miR-17-5p significantly promoted the proliferation (P＜0.05 ), invasion (P＜0.01 ) and migration (P＜0.05) of lung adenocarcinoma cells, and inhibited apoptosis (P＜0.01 ). There was a direct binding site between miR-17-5p and 3'UTR of PKD2, and overexpression of miR-17-5p significantly promoted the expression of PKD2 at both mRNA and protein levels (P＜0.01). Conclusion: PKD2 was the direct target gene of miR-17-5p, and miR-17-5p affected the biological behavior of lung adenocarcinoma cells by regulating PKD2.
[1] Sayin VI, LeBoeuf SE, Papagiannakopoulos T. Targeting metabolic bottlenecks in lung cancer[J].Trends Cancer, 2019,5(8): 457-459.
[2] Herbst RS, Morgensztern D, Boshoff C. The biology and management of non-small cell lung cancer[J].Nature,2018,553(7689): 446-454.
[3] Michlewski G, Cáceres JF. Post-transcriptional control of miRNA biogenesis[J].RNA, 2019,25(1): 1-16.
[4] Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function[J].Nucleic Acids Res, 2019,47(D1): D155-D162.
[5] Lee TJ, Yuan X, Kerr K, et al. Strategies to modulate microRNA functions for the treatment of cancer or organ injury[J].Pharmacol Rev,2020,72(3): 639-667.
[6] Zhang Y, Zhang Y, Yin Y, et al. Detection of circulating exosomal miR-17-5p serves as a novel non-invasive diagnostic marker for non-small cell lung cancer patients[J].Pathol Res Pract, 2019,215(8): 152466.
[7] Zhang G, An X, Zhao H, et al. Long non-coding RNA HNF1A-AS1 promotes cell proliferation and invasion via regulating miR-17-5p in non-small cell lung cancer[J].Biomed Pharmacother, 2018,98: 594-599.
[8] Wang Y, Xu W, Wang Y, et al. miR-17-5p promotes migration and invasion in breast cancer cells by repressing netrin 4[J].Int J Clin Exp Pathol, 2019,12(5): 1649-1657.
[9] Zhao X, Xu Y, Sun X, et al. miR-17-5p promotes proliferation and epithelial-mesenchymal transition in human osteosarcoma cells by targeting SRC kinase signaling inhibitor 1[J].J Cell Biochem, 2019,120(4): 5495-5504.
[10] Zhu Y, Gu J, Li Y, et al. MiR-17-5p enhances pancreatic cancer proliferation by altering cell cycle profiles via disruption of RBL2/E2F4-repressing complexes[J].Cancer Lett, 2018,412: 59-68.
[11] Chen Y, Zhou X, Huang C, et al. LncRNA PART1 promotes cell proliferation and progression in non-small-cell lung cancer cells via sponging miR-17-5p[J].J Cell Biochem, 2021,122(3-4): 315-325.
[12] 罗凯,黎谢梦丹,郑国沛,等. MiR-17-5p靶向PTEN促进非小细胞肺癌吉非替尼耐药[J].实用医学杂志,2019,35(11): 1717-1721,1726.
[13] Farooqi AA, Fuentes-Mattei E, Fayyaz S, et al. Interplay between epigenetic abnormalities and deregulated expression of microRNAs in cancer[J].Semin Cancer Biol, 2019,58: 47-55.
[14] 汪国华. miRNA及转录因子结合位点预测与调控功能分析[D].哈尔滨:哈尔滨工业大学,2009.
[15] Lu J, Clark AG. Impact of microRNA regulation on variation in human gene expression[J].Genome Res, 2012,22(7): 1243-1254.
[16] Dellago H, Bobbili MR, Grillari J. MicroRNA-17-5p: At the crossroads of cancer and aging - a mini-review[J].Gerontology, 2017,63(1): 20-28.
[17] 刘鹏飞. 过表达microRNA-17-5p对结肠癌HCT116细胞生物学功能的影响[D].石家庄:河北医科大学,2019.
[18] Habashy DA, El Tayebi HM, Fawzy IO, et al. Interplay between microRNA-17-5p, insulin-like growth factor-II through binding protein-3 in hepatocellular carcinoma[J].World J Hepatol, 2016,8(23): 976-984.
[19] Ming He J, Liu PY, Wang J. MicroRNA-17-5p regulates the growth, migration and invasion of the human osteosarcoma cells by modulating the expression of PTEN[J].J BUON, 2020,25(2): 1028-1034.
[20] Geng X, Sun Y, Fu J, et al. MicroRNA-17-5p inhibits thyroid cancer progression by suppressing Early growth response 2 (EGR2)[J].Bioengineered, 2021,12(1): 2713-2722.
[21] Sun W, Cui J, Ge Y, et al. Tumor stem cell-derived exosomal microRNA-17-5p inhibits anti-tumor immunity in colorectal cancer via targeting SPOP and overexpressing PD-L1[J].Cell Death Discov, 2022,8(1): 223.
[22] 赵云东,宋玉琴,刘娜,等. MiR-543对胶质瘤SHG-44细胞侵袭、迁移的影响及机制研究[J].宁夏医科大学学报,2018,40(07):770-775.
[23] 李福利,夏咸军,刘洪,等. PKD2与GOLPH3在胆囊癌组织中的表达及意义[J].现代肿瘤医学,2021,29(13):2296-2300.
[24] 陈和萍,俞力军,乐湘华,等. 胃癌组织中FOXP1和PKD2的表达及临床意义[J].胃肠病学和肝病学杂志, 2020,29(05):520-524.
[25] 张珊珊,杜易. FOXP1和PKD2在子宫内膜癌中的表达及临床意义[J].中国妇幼保健, 2020,35(10):1917-1920.
[26] 庞兆飞. 蛋白激酶D2(PKD2)促进肺腺癌上皮间质转化的作用及机制研究[D].济南:山东大学,2019.
