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目的：骨肉瘤源于骨骼系统，由于其分子机制复杂，目前治疗手段有限。丝氨酸/苏氨酸蛋白激酶有丝分裂基因A相关表达激酶6(nerve in mitosis gene A-related expressed kinase 6, NEK6)可以调控细胞周期和多种致癌信号通路。本研究旨在分析NEK6在骨肉瘤中的表达及对其转移和细胞凋亡的调控作用。方法：收集骨肉瘤患者的肿瘤组织及正常组织，用实时定量PCR(real-time-quantitative PCR, RT-qPCR)检测NEK6的mRNA表达，以及在骨肉瘤细胞MG-63和Saos-2中，用RT-qPCR、Western Blot和免疫荧光检测NEK6的表达，进一步用siRNA抑制NEK6表达，通过CCK-8、细胞划痕、Transwell和流式细胞术等方法观察对骨肉瘤细胞增殖、迁移、侵袭和凋亡的影响。用Western Blot检测STAT3磷酸化水平及凋亡、转移相关蛋白的表达。结果：NEK6在骨肉瘤组织及细胞系中均高表达；siRNA抑制NEK6表达后，骨肉瘤细胞的增殖、迁移和侵袭得到抑制并促进了细胞凋亡。STAT3磷酸化水平增高，转移相关基因基质金属蛋白酶(matrix metallopeptidase, MMP)-2和MMP-9表达下降，促凋亡基因Bax上调，抑凋亡基因Bcl-2下调。结论 ：NEK6是骨肉瘤潜在的促癌基因，抑制NEK6可以抑制骨肉瘤细胞的增殖、迁移和侵袭并促进凋亡。NEK6可以通过激活STAT3信号通路促进骨肉瘤的转移和抑制凋亡，用siRNA等方法抑制NEK6的策略可能是治疗骨肉瘤的有效方法。
Abstract：Objective: Osteosarcoma originates from the skeletal system, and due to its complex molecular mechanism, current treatment options are limited. Nerve in mitosis gene A-related expressed kinase 6(NEK6) is a serine/threonine protein kinase that can regulate the cell cycle and various oncogenic signaling pathways. The aim of this study is to investigate NEK6 expression in osteosarcoma and its regulatory effects on metastasis and cell apoptosis. Methods: The tumor tissues from osteosarcoma patients and control normal tissues were collected, NEK6 mRNA levels were detected using real-time-quantitative PCR(RT-qPCR), and NEK6 expression in MG-63 and Saos-2 cells were detected by using RT-qPCR, Western Blot, and immunofluorescence staining. Further NEK6 expression were inhibited using specific siRNA, and the effects on osteosarcoma cell proliferation, migration, invasion, and apoptosis were observed by using CCK-8, cell scratch, Transwell, and flow cytometry methods. Western Blot was used to detect the phosphorylation levels of STAT3, the levels of apoptosis and metastasis related proteins. Results: NEK6 was highly expressed in osteosarcoma tissues and cells; after inhibiting NEK6 expression with siRNA,the proliferation, migration and invasion of osteosarcoma cells were suppressed but apoptosis was promoted. The phosphorylation levels of STAT3 increased, the expression of metastasis related genes matrix metallopeptidase(MMP)-2 and MMP-9 decreased,the pro-apoptotic gene Bax was upregulated, and the apoptosis inhibitory gene Bcl-2 was downregulated. Conclusion: NEK6 is a potential oncogene in osteosarcoma. Inhibiting NEK6 expression can inhibit the proliferation, migration and invasion of osteosarcoma cells and promote apoptosis. NEK6 can promote the metastasis and inhibit apoptosis of osteosarcoma by activating the STAT3 signaling pathway. The strategy of inhibiting NEK6 using siRNA and other methods may be effective treatments for osteosarcoma.
[1]FENG L,CHEN Y,JIN W.Research progress on cancerassociated fibroblasts in osteosarcoma[J].Oncol Res,2025,33(5):1091-1103.
[2]KEEGAN T H M,ABRAHÃO R,ALVAREZ E M.Survival trends among adolescents and young adults diagnosed with cancer in the United States:comparisons with children and older adults[J].J Clin Oncol,2024,42(6):630-641.
[3]LI H,LI J,ZHANG Y,et al.The NIMA-related kinase family and cancer[J].Front Oncol,2025,15:1556917.
[4]FOLAHAN J T,BARABUTIS N.NEK kinases in cell cycle regulation,DNA damage response,and cancer progression[J].Tissue Cell,2025,94:102811.
[5]PANCHAL N K,EVAN PRINCE S.The NEK family of serine/threonine kinases as a biomarker for cancer[J].Clin Exp Med,2023,23(1):17-30.
[6]YU Y,SHEN T,ZHONG X,et al.NEK6 is an injuryresponsive kinase cooperating with STAT3 in regulation of reactive astrogliosis[J].Glia,2022,70(2):273-286.
[7]HE Z,NI X,XIA L,et al.Overexpression of NIMA-related kinase 6(NEK6) contributes to malignant growth and dismal prognosis in Human Breast Cancer[J].Pathol Res Pract,2018,214(10):1648-1654.
[8]PAVAN I C B,BASEI F L,SEVERINO M B,et al.NEK6 regulates redox balance and DNA damage response in DU-145 prostate cancer cells[J].Cells,2023,12(2):256.
[9]SU L,ZHAO D,ZHOU C,et al.NEK6 accelerates hepatocellular carcinoma progression and glycolysis through ubiquitination of TCP10L[J].Crit Rev Eukaryot Gene Expr,2025,35(4):1-13.
[10]GAO S,SU M,BIAN T T,et al.NEK6 functions as an oncogene to promote the proliferation and metastasis of ovarian cancer[J].J Cancer,2025,16(4):1335-1346.
[11]HONG Z,CHEN Z,PAN J,et al.micro RNA-323a-3p negatively regulates NEK6 in colon adenocarcinoma cells[J].JOncol,2022,2022:7007718.
[12]LIU Y,FU W,YIN F,et al.mi R-141-3p suppresses development of clear cell renal cell carcinoma by regulating NEK6[J].Anticancer Drugs,2022,33(1):e125-e133.
[13]BEAUCHEMIN M P,RAGHUNATHAN R R,ACCORDINOM K,et al.New persistent opioid use among adolescents and young adults with sarcoma[J].Cancer,2022,128(14):2777-2785.
[14]TODOSENKO N,KHLUSOV I,YUROVA K,et al.Signal pathways and micro RNAs in osteosarcoma growth and the dual role of mesenchymal stem cells in oncogenesis[J].Int J Mol Sci,2023,24(10):8993.
[15]CHEN L,LU H,BALLOUT F,et al.Targeting NEK kinases in gastrointestinal cancers:insights into gene expression,（上接第523页）function,and inhibitors[J].Int J Mol Sci,2025,26(5):1992.
[16]DUTT P,HAIDER N,MOUAAZ S,et al.β-catenin turnover is regulated by Nek10-mediated tyrosine phosphorylation in A549 lung adenocarcinoma cells[J].Proc Natl Acad Sci USA,2024,121(19):e2300606121.
[17]ZHANG Y,WANG W,WANG Y,et al.NEK2 promotes hepatocellular carcinoma migration and invasion through modulation of the epithelial-mesenchymal transition[J].Oncol Rep,2018,39(3):1023-1033.
[18]SABIR S R,SAHOTA N K,JONES G D D,et al.Loss of Nek11 prevents G2/M arrest and promotes cell death in HCT116 colorectal cancer cells exposed to therapeutic DNAdamaging agents[J].PLo S One,2015,10(10):e0140975.
[19]DE CASTRO FEREZIN C,LIM KAM SIAN T C C,WU YJ,et al.Identification of biological pathways and processes regulated by NEK5 in breast epithelial cells via an integrated proteomic approach[J].Cell Commun Signal,2022,20(1):197.
[20]YAN Z,QU J,LI Z,et al.NEK7 promotes pancreatic cancer progression and its expression is correlated with poor prognosis[J].Front Oncol,2021,11:705797.
[21]PANCHAL N K,MOHANTY S,PRINCE S E.NIMA-related kinase-6(NEK6) as an executable target in cancer[J].Clin Transl Oncol,2023,25(1):66-77.
基本信息:
DOI：10.16424/j.cnki.cn32-1807/r.2025.06.002
中图分类号:R738.1
引用信息:
[1]朱敏,严晓玲,孙郁雨,等.激酶NEK6调控STAT3信号通路促进骨肉瘤细胞转移并抑制凋亡[J].南通大学学报(医学版),2025,45(06):519-523+529.DOI:10.16424/j.cnki.cn32-1807/r.2025.06.002.
基金信息:
南通市自然科学基金; 社会民生科技计划项目(MSZ2023126)