ZNF750 facilitates carcinogenesis via promoting the expression of long non-coding RNA CYTOR and influences pharmacotherapy response in colon adenocarcinoma

ZNF750 facilitates carcinogenesis via promoting the expression of long non-coding RNA CYTOR and influences pharmacotherapy response in colon adenocarcinoma 锌指蛋白750通过促进长链非编码RNA CYTOR 表达促进结直肠癌癌变并影响药物治疗反应 1 first-author 2 3 4 corresp Jiabian LIAN, faster_lian@xmu.edu.cn Jiabian LIAN, faster_lian@xmu.edu.cn faster_lian@xmu.edu.cn https://orcid.org/0000-0003-0934-6633 Xiamen Cell Therapy Research Center, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China Xiamen Cell Therapy Research Center, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China 厦门大学附属第一医院，厦门市细胞治疗研究中心，中国厦门，361000 厦门大学附属第一医院，厦门市细胞治疗研究中心，中国厦门，361000 Department of Colorectal Surgery, the First Affiliated Hospital of Fujian Medical University, Fuzhou 350004, China Department of Colorectal Surgery, the First Affiliated Hospital of Fujian Medical University, Fuzhou 350004, China 福建医科大学附属第一医院结直肠外科，中国福州，350004 福建医科大学附属第一医院结直肠外科，中国福州，350004 Department of Gastrointestinal Surgery, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China Department of Gastrointestinal Surgery, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China 厦门大学附属第一医院胃肠肿瘤外科，中国厦门，361000 厦门大学附属第一医院胃肠肿瘤外科，中国厦门，361000 Department of Clinical Laboratory, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China Department of Clinical Laboratory, the First Affiliated Hospital of Xiamen University, Xiamen 361000, China 厦门大学附属第一医院检验科，中国厦门，361000 厦门大学附属第一医院检验科，中国厦门，361000 目的： 2 探讨锌指蛋白750在调控长链非编码RNA CYTOR 表达进而影响结直肠癌发生发展中的相关表型和机制及其影响药物应答和预后的潜在临床意义。 创新点： 2 聚焦锌指蛋白750在结肠腺癌中的促癌机制，并在其对 CYTOR 表达的影响及锌指蛋白750- CYTOR 与结肠腺癌临床相关分子病理特征和药物疗效预测方面展开研究。 方法： 2 在分子水平，利用蛋白质免疫印迹（western blotting）、染色质免疫沉淀-定量聚合酶链反应（ChIP-qPCR）、转录组测序（RNA-seq）、逆转录-定量聚合酶链反应（RT-qPCR）等技术手段对ZNF750和 CYTOR 在结肠腺癌细胞系中的表达进行分析；在细胞水平，通过使用克隆形成（colony formation）、小室迁移（transwell）等方法评估ZNF750对结肠腺癌增殖、侵袭、转移功能表型的影响；在动物水平，评估在免疫缺陷的裸鼠模型中，ZNF750对人结肠腺癌细胞系皮下移植瘤生长能力的影响；对TCGA和GEO数据库中结肠腺癌患者ZNF750- CYTOR 的表达量、表达相关性进行生物信息学分析，并评估在对患者预后、肿瘤免疫浸润和常规化疗药物疗效预测中的价值。 结论： 2 锌指蛋白750正向调节 CYTOR 表达，且锌指蛋白750- CYTOR 可促进结肠腺癌细胞的恶性表型。对TCGA或GEO结肠腺癌队列的研究结果显示，锌指蛋白750- CYTOR 轴与临床特征间存在三个重要相关性：首先，锌指蛋白750- CYTOR 轴在癌变和癌症进展过程上调；其次，锌指蛋白750- CYTOR 与肿瘤浸润淋巴细胞（TILs）呈正相关，与免疫检查点基因的表达有关；第三，锌指蛋白750- CYTOR 可作为结肠腺癌药物治疗中亚叶酸钙或奥沙利铂等常规化疗药物的反应预测因子。 锌指蛋白750 结肠腺癌 长链非编码RNA CYTOR 化学药物治疗 The epidermal cell differentiation regulator zinc finger protein 750 (ZNF750) is a transcription factor containing the Cys2His2 (C2H2) domain, the zinc finger structure of which is located at the N-terminal 25‍‍–‍46 amino acids of ZNF750. It can promote the expression of differentiation-related factors while inhibiting the expression of progenitor cell-related genes. ZNF750 is directly regulated by p63 (encoded by the TP63 gene, belonging to the TP53 superfamily). The Krüppel-like factor 4 (KLF4), repressor element-1 (RE-1)‍-silencing transcription factor (REST) corepressor 1 (RCOR1), lysine demethylase 1A (KDM1A), and C-terminal-binding protein 1/2 (CTBP1/2) chromatin regulators cooperate with ZNF750 to repress epidermal progenitor genes and activate the expression of epidermal terminal differentiation genes ( Sen et al., 2012 Sen et al., 2012 ; Boxer et al., 2014 Boxer et al., 2014 ). Besides, ZNF750 and the regulatory network composed of bone morphogenetic protein (BMP) signaling pathway, long non-coding RNAs (lncRNAs) (anti-differentiation non-coding RNA (ANCR) and tissue differentiation-inducing non-protein coding RNA (TINCR)), musculoaponeurotic fibrosarcoma oncogene (MAF)/MAF family B (MAFB), grainy head-like 3 (GRHL3), and positive regulatory domain zinc finger protein 1 (PRDM1) jointly promote epidermal cell differentiation ( Sen et al., 2012 Sen et al., 2012 ). Studies have shown that ZNF750 has an inhibitory effect on various tumors such as esophageal squamous cell carcinoma, oral squamous cell carcinoma, and nasopharyngeal carcinoma ( Hao et al., 2016 Hao et al., 2016 ; Hazawa et al., 2017 Hazawa et al., 2017 ; Zhang et al., 2018 Zhang et al., 2018 ). The compromised level of ZNF750 in squamous cell carcinoma cells was associated with poor clinical prognosis. In addition, ZNF750 promotes the expression of lncRNA TINCR ( Hazawa et al., 2017 Hazawa et al., 2017 ). Zhang et al. (2018) Zhang et al. (2018) confirmed that: (1) N 6 -methyladenosine (m 6 A) modification maintained the low level of ZNF750 in nasopharyngeal carcinoma; (2) ZNF750 directly regulates the transcription of fibroblast growth factor 14 (FGF14); and (3) the ZNF750-FGF14 signal axis inhibits the growth of nasopharyngeal carcinoma cells by promoting apoptosis. However, studies focused on the mechanistic functions of ZNF750 in colorectal adenocarcinoma have been very limited ( Qu et al., 2020 Qu et al., 2020 ). LncRNA cytoskeleton regulator ( CYTOR ), also known as LINC00152 , is a widely reported differentially expressed lncRNA in cancers of different body parts such as the stomach, liver, colon, lung, gallbladder, kidney, breast, and oral cavity ( Xu et al., 2017 Xu et al., 2017 ). Studies have shown that lncRNA CYTOR can promote the malignant phenotype of cancer cells ( Deng et al., 2017 Deng et al., 2017 ), and facilitate the compromising of medication sensitivity ( Yue et al., 2016 Yue et al., 2016 , 2018 2018 ; Bian et al., 2017 Bian et al., 2017 ). It was also related to the poor prognosis of patients. CYTOR can be transcriptionally activated by transcription factor 3 (TCF3) ( Zheng et al., 2019 Zheng et al., 2019 ), transcription factor Yin Yang 1 (YY1) ( Shen et al., 2019 Shen et al., 2019 ), specificity protein 1 (SP1) ( Cai et al., 2016 Cai et al., 2016 ), and Yes-associated protein 1 (YAP1) ( Ou et al., 2020 Ou et al., 2020 ). CYTOR competes with various microRNAs (miRNAs) such as miR-139-5p ( Bian et al., 2017 Bian et al., 2017 ), miR-1182 ( Zheng et al., 2019 Zheng et al., 2019 ), miR-497 ( Sun et al., 2019 Sun et al., 2019 ), miR-162 ( Cai et al., 2018 Cai et al., 2018 ), miR-125b ( Chen et al., 2018 Chen et al., 2018 ), miR-193a/b-3p ( Ma et al., 2018 Ma et al., 2018 ; Wang et al., 2019 Wang et al., 2019 ), miR-138 ( Cai et al., 2017 Cai et al., 2017 ), and miR-103a-3p ( Yu MJ et al., 2017 Yu MJ et al., 2017 ) as competing endogenous RNA (ceRNA) to regulate notch receptor 1 (NOTCH1) ( Bian et al., 2017 Bian et al., 2017 ), cyclin-dependent kinase 14 (CDK14) ( Zheng et al., 2019 Zheng et al., 2019 ), brain-derived neurotrophic factor (BDNF) ( Sun et al., 2019 Sun et al., 2019 ), protein kinase B (PKB)‍-‍β (AKT2)/nuclear factor-κB (NF-κB) ( Cai et al., 2018 Cai et al., 2018 ), myeloid cell leukemia-1 (MCL-1) ( Chen et al., 2018 Chen et al., 2018 ), cyclin D1 (CCND1) ( Ma et al., 2018 Ma et al., 2018 ), E26 transformation-specific 1 (ETS1) ( Wang et al., 2019 Wang et al., 2019 ), hypoxia-inducible factor-‍1α (HIF-‍1α) ( Cai et al., 2017 Cai et al., 2017 ), FEZ family zinc finger 1 (FEZF1) ( Yu MJ et al., 2017 Yu MJ et al., 2017 ), and epidermal growth factor receptor-4 (ERBB4) ( Yue et al., 2016 Yue et al., 2016 ). CYTOR interacts with thrombospondin-1 (THBS1) ( Xia et al., 2014 Xia et al., 2014 ), SET and MYND domain protein 2 (SMYD2) ( Shan et al., 2017 Shan et al., 2017 ), β‍-catenin ( Shan et al., 2017 Shan et al., 2017 ; Yue et al., 2018 Yue et al., 2018 ), enhancer of zeste homolog 2 (EZH2) ( Chen et al., 2017 Chen et al., 2017 ; Deng et al., 2017 Deng et al., 2017 ; Wang et al., 2017 Wang et al., 2017 ), lysine-specific demethylase 1 (LSD1) ( Wang et al., 2017 Wang et al., 2017 ), trimethylated histone H3 lysine 27 (H3K27me3) ( Wang et al., 2017 Wang et al., 2017 ), and other transcriptional and epigenetic factors to orchestrate downstream gene expression. CYTOR binds with the interleukin-24 (IL-24) ( Chen et al., 2017 Chen et al., 2017 ) or epithelial cell adhesion molecule (EpCAM) ( Ji et al., 2015 Ji et al., 2015 ) promoter to regulate their expression. According to the above results, CYTOR regulates signal pathways such as Wnt/β‍-‍catenin ( Shan et al., 2017 Shan et al., 2017 ; Yue et al., 2018 Yue et al., 2018 ), NF-‍‍κB ( Wang et al., 2018 Wang et al., 2018 ), phosphoinositide-3-kinase (PI3K)/AKT ( Cai et al., 2016 Cai et al., 2016 ; Yu MJ et al., 2017 Yu MJ et al., 2017 ), and mammalian target of rapamycin (mTOR) ( Ji et al., 2015 Ji et al., 2015 ), which play vital roles in cancer biology ( Yu Y et al., 2017 Yu Y et al., 2017 ; Xu et al., 2019 Xu et al., 2019 ; Ou et al., 2020 Ou et al., 2020 ). However, the role of lncRNA CYTOR in colorectal cancer (CRC) is still controversial. Studies have shown that the level of CYTOR increases in colon malignant tissues, and its expression level is positively correlated with the clinical stage of the tumor and lymph node metastasis ( Yue et al., 2016 Yue et al., 2016 , 2018 2018 ; Yu Y et al., 2017 Yu Y et al., 2017 ). Further studies have found that CYTOR not only enhances the tumorigenicity of colon cancer cells, but also is related to CRC chemotherapy resistance by interacting with β‍-catenin protein and forming a positive feedback loop ( Yue et al., 2018 Yue et al., 2018 ). The YAP1/ CYTOR axis facilitates the progression of CRC ( Ou et al., 2020 Ou et al., 2020 ). CYTOR works as ceR binding to miR-193a-3p, upregulating the expression of the ERBB4 oncogene, which further activates the PKB/AKT signaling pathway and reduces the sensitivity of colon cancer cells to chemotherapy drugs ( Yue et al., 2016 Yue et al., 2016 ). The CYTOR -nucleolin-Sam68/KHDRBS1 heterotrimeric complex activates the NF‍-‍‍κB pathway and epithelial-mesenchymal transition (EMT) to potentiate CRC progression ( Wang et al., 2018 Wang et al., 2018 ). Meanwhile, some studies have shown that upregulating the expression of CYTOR decreases the activity of colon cancer cells and increases apoptosis, suggesting that CYTOR may suppress tumorigenesis in colon cancer cells ( Zhang et al., 2016 Zhang et al., 2016 ). These studies indicated that CYTOR may play a dual role of tumor promoter gene and tumor suppressor gene. Further research should be untaken to reveal the functional role of CYTOR in colon cancer progression and the underlying mechanism. In the present study, we found that ZNF750 expression was higher in colon adenocarcinoma (COAD) tissue than in the normal colonic mucosal tissue in The Cancer Genome Atlas (TCGA)-COAD cohort ( Fig. 1 Fig. 1 a). The receiver operating characteristic (ROC) curve analysis showed that ZNF750 is a good predictor of COAD ( Fig. 1 Fig. 1 b). Low ZNF750 expression was a good prognostic marker in both the TCGA-COAD and GSE39582 datasets ( Fig. 1 Fig. 1 c). In order to assess the function of ZNF750 in CRC progression, we detected the expression of ZNF750 using western blotting in various COAD cells, among which the human colon cancer cell line SW620 showed the highest ZNF750 expression level, and human colon cancer cell lines (HCT116, CACO2, and SW480) showed moderate expression levels ( Fig. 2 Fig. 2 a). Next, ZNF750 expression was either increased or reduced in HCT116 cell lines using overexpression constructs or short hairpin RNA (shRNA) constructs. The expression levels of ZNF750 were effectively altered ( Fig. 2 Fig. 2 b). The cell proliferation assay revealed that ZNF750 overexpression improved the growth speed, whereas ZNF750 silencing slowed it down (Figs. 2c 2c and 2d 2d ). Furthermore, the colony formation assays demonstrated that ZNF750 overexpression had an augmentation effect, while ZNF750 silencing exerted a weakening effect on the cloning formation and proliferation capacity of CRC cells ( Fig. 2 Fig. 2 e). The transwell assays indicated the increased migration and invasion of HCT116 cells with ZNF750 overexpression, and reduced migration and invasion in HCT116 cells with ZNF750 silencing (Figs. 2g 2g and 2h 2h ). Furthermore, we injected HCT116 ZNF750-silencing cells or control cells subcutaneously into nude mice. The tumor volume measurements showed that the ZNF750-silencing group generated smaller tumors compared with the controls ( Fig. 2 Fig. 2 f; n =3, where one failed to form a subcutaneous tumor). In order to explore the molecular mechanism by which ZNF750 directly promotes the malignant CRC type, we performed RNA sequencing (RNA-seq) to investigate the transcription level alterations occurring along with ZNF750 overexpression. The results showed that the expression levels of multiple genes changed with the alteration of ZNF750, among which 277 genes were upregulated and 291 genes were downregulated in HCT116 cells ( P < 0.05). According to Gene Set Enrichment Analysis (GSEA) in the HCT116 cell line, ZNF750 regulates subsets of genes highly enriched in the tumor necrosis factor-α (TNF-‍α) signaling via NF-‍κB, EMT, hypoxia, transforming growth factor-‍β (TGF-‍β) signaling, and hypoxia (Fig. S1a). The GSEA results of TCGA-COAD dataset were similar to those of RNA-seq in the HCT116 cell line (Fig. S1b). Since the RNA-seq data only contain the information of messenger RNA (mRNA) with poly-A tail, we further analyzed the expression of ZNF750 and lncRNA in the TCGA-COAD datasets. ZNF750 is positively correlated with 15 lncRNAs and negatively correlated with four lncRNAs in the TCGA-COAD datasets ( Fig. 3 Fig. 3 a). CYTOR is one of lncRNAs positively correlated with ZNF750 in the TCGA-COAD dataset ( P < 0.001; Fig. 3 Fig. 3 b). The chromatin immunoprecipitation (ChIP)-quantitative real-time polymerase chain reaction (qPCR) results confirmed the binding of ZNF750 in the gene region of CYTOR in SW620 cells, which has been detected in the ChIP-seq data numbered SRR1287820 (Figs. 3c 3c and 3d 3d ). Furthermore, the reverse transcription (RT)‍-qPCR experiment confirmed that the expression of lncRNA- CYTOR decreased with ZNF750 silencing in HCT116 cells ( Fig. 3 Fig. 3 e). Taken together, these results revealed that ZNF750 transcriptionally regulates CYTOR expression. In order to reveal the possible role of the ZNF750- CYTOR axis in COAD development, the difference in the RNA expression of CYTOR in COAD and normal tissues was evaluated. CYTOR expression was higher in COAD tissue than in paired normal tissue in accordance with ZNF750 ( Fig. 4 Fig. 4 a). The ROC curve analysis revealed that CYTOR is a good predictor of COAD ( Fig. 4 Fig. 4 b). Although low ZNF750 expression was a good prognostic marker, there was no significant survival advantage in the low CYTOR expression group in the TCGA-COAD dataset ( Fig. 4 Fig. 4 c; left panel). However, when classified by stages, low CYTOR expression was a good prognostic marker for patients in stages I+II in the TCGA-COAD dataset ( Fig. 4 Fig. 4 c; middle panel). The RNA-seq results mentioned above revealed that the TNF-α_SIGNALING_VIA_NF-‍κB is the most significant GSEA item influenced by ZNF750 overexpression in HCT116 cells. Moreover, NF-‍κB signaling is a vital regulatory pathway involved in tumor growth and at the same time is one of the linking hubs between chronic inflammation and neoplastic development. Prompted by this finding, the potential immune modulation roles of ZNF750 and CYTOR in TCGA-COAD data were analyzed. In COAD, the abundance of cluster of differentiation 4-positive (CD4 + ) cells, macrophages, neutrophils, and dendritic cells correlated positively with ZNF750; CD8 + cells, macrophages, neutrophils, and dendritic cells correlated positively with CYTOR expression (Fig. S2a). ZNF750 and CYTOR expression levels were both positively linked to immune checkpoint genes, including programmed death-ligand 1 ( PD-L1 ), CD86 , PD-L2 , and cytotoxic T-lymphocyte antigen-4 ( CTLA4 ) (Fig. S2b). These genes are immune checkpoints and efficacy-related influencers of immunotherapy. The assessment of the potential roles of ZNF750 and CYTOR in immunotherapy requires further investigation by new cohorts using immunotherapy. In order to explore the possible roles of altered ZNF750- CYTOR expression in response to guideline-recommended medication, survival analysis was applied to validate the survival differences between patients treated with specific medications. For those who underwent leucovorin or oxaliplatin therapy, the death risk in the high ZNF750/ CYTOR group was smaller than that in the low ZNF750/ CYTOR group ( Figs. 5 Figs. 5 a‍–‍5c). These results indicated that ZNF750- CYTOR may serve as a prognostic marker of specific chemotherapy. In squamous cell carcinoma, ZNF750 is a lineage-specific tumor suppressive transcription factor. However, little evidence has shown its role in COAD or as a therapeutic target ( Wen et al., 2021 Wen et al., 2021 ). In the present study, we demonstrated that ZNF750 may promote carcinogenesis and progression either through modulating ZNF750 expression in vitro and in vivo, or the correlation analysis of ZNF750 and clinical pathological studies. Evidence shows that lncRNA modulates cancer progression ( Chen and Shen, 2020 Chen and Shen, 2020 ). LncRNA CYTOR plays a crucial role in COAD development, which has been reviewed previously. It is now clear that tumors orchestrate certain immune checkpoint pathways as a major mechanism of immune resistance. A plethora of immune checkpoint regulators have been identified ( Pardoll, 2012 Pardoll, 2012 ), among which two have been studied the most intensively: CTLA4 binding to CD80 or CD86 and programmed cell death protein 1 (PDCD1, also called PD1) binding to CD274 (also known as PD-L1) or PDCD1 ligand 2 (PDCD1LG2). Microsatellite instability-high (MSI-H), tumor mutational burden (TMB), and PD-L1 simultaneously exist in 12.8% of CRC cases, which is much higher than that in all cancers (2.9%) mentioned in European Society of Medical Oncology (ESMO) recommendations. Besides, TMB and MSI-H show strong correlation in 44.2% of CRC ( Luchini et al., 2019 Luchini et al., 2019 ). These clinical data indicate tremendous possibilities of immunological therapy in CRC. The ZNF750- CYTOR axis shows strong correlation with currently known immune therapy indicators (Fig. S2), prompting the notion that ZNF750- CYTOR participates in immune therapy. Radical surgical resection and adjuvant therapy are the primary treatment methods for advanced COAD. In the present study, patients with a high expression of ZNF750/ CYTOR had different prognosis results depending on the received therapy, which indicates that the prediction value of whether ZNF750/ CYTOR is a good potential medication target or therapeutic efficacy predictor varies by the specific kind of drug applied. So far, no study has shown that ZNF750 can serve as a therapeutic target for tumors. Hence, further work should be undertaken to explore whether ZNF750 is an appropriate target for the therapy of different kinds of cancer. This study has certain limitations. The exact mechanisms of ZNF750- CYTOR axis affecting tumor-infiltrating lymphocyte (TIL) infiltration and the modulating efficacy of immune checkpoint blocker regents require further experiments to explore. In addition, new immunotherapy information for COAD to verify the role of ZNF750- CYTOR axis through additional clinical experiments would prove useful. Beyond these limitations, this is the first study to demonstrate that ZNF750- CYTOR promotes tumor malignant phenotypes and influences chemotherapy response. In conclusion, this paper has revealed that ZNF750 positively regulates CYTOR expression through in vitro experiments. ZNF750- CYTOR promotes the malignant phenotypes in COAD cells. Furthermore, studies based on TCGA or Gene Expression Omnibus (GEO) COAD cohorts have indicated three vital relevancies between the ZNF750- CYTOR axis and clinical features: first, the ZNF750- CYTOR axis is upregulated during carcinogenesis and cancer progression; second, ZNF750- CYTOR positively correlates with the TILs and is related to the expression of immune checkpoint genes; third, ZNF750- CYTOR may be a pharmacotherapy response predictor of COAD for several kinds of conventional chemotherapy. Materials and methods 1 Detailed methods are provided in the electronic supplementary materials of this paper. References Bian ZH , Zhang JW , Li M , et al. , 2017 . Long non-coding RNA LINC00152 promotes cell proliferation, metastasis, and confers 5-FU resistance in colorectal cancer by inhibiting miR-139-5p . 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A positive feed-forward loop between LncRNA-CYTOR and Wnt/β‍-catenin signaling promotes metastasis of colon cancer . Mol Ther , 26 ( 5 ): 1287 - 1298 . https://doi.org/10.1016/j.ymthe.2018.02.024 https://doi.org/10.1016/j.ymthe.2018.02.024 A positive feed-forward loop between LncRNA-CYTOR and Wnt/β‍-catenin signaling promotes metastasis of colon cancer Zhang PP , He QP , Lei Y , et al. , 2018 . m 6 A-mediated ZNF750 repression facilitates nasopharyngeal carcinoma progression . Cell Death Dis , 9 ( 12 ): 1169 . https://doi.org/10.1038/s41419-018-1224-3 https://doi.org/10.1038/s41419-018-1224-3 m6A-mediated ZNF750 repression facilitates nasopharyngeal carcinoma progression Zhang YH , Fu J , Zhang ZJ , et al. , 2016 . LncRNA-LINC00152 down-regulated by miR-376c-3p restricts viability and promotes apoptosis of colorectal cancer cells . Am J Transl Res , 8 ( 12 ): 5286 - 5297 . LncRNA-LINC00152 down-regulated by miR-376c-3p restricts viability and promotes apoptosis of colorectal cancer cells Zheng LL , Hu N , Zhou XZ , 2019 . TCF3-activated LINC00152 exerts oncogenic role in osteosarcoma through regulating miR-1182/CDK14 axis . Pathol Res Pract , 215 ( 2 ): 373 - 380 . https://doi.org/10.1016/j.prp.2018.12.031 https://doi.org/10.1016/j.prp.2018.12.031 TCF3-activated LINC00152 exerts oncogenic role in osteosarcoma through regulating miR-1182/CDK14 axis Lu XIA performed the experimental research and data analysis, wrote and edited the manuscript. Hexin LIN performed the data analysis. Yanming ZHOU performed the establishment of animal models. Jiabian LIAN contributed to the study design, writing and editing of the manuscript. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data. 10.1631/jzus.B2100939 1673-1581（2022）07-0587-10 This work was supported by the National Natural Science Foundation of China (No. 81802268), the Fujian Provincial Department of Science and Technology (Nos. 2020J05298 and 2021J011364), the Fujian Province Health and Family Planning (No. 2019-CXB-30), and the Xiamen City Department of Health (No. 3502Z20214ZD1006), China. 2021-11-15 2022-03-21 2022-07-15 2022-12-27 2022-05-05 Copyright © Zhejiang University and Springer-Verlag GmbH Germany, part of Springer Nature 2022 2022 浙江大学出版社 Lu XIA, Hexin LIN, Yanming ZHOU and Jiabian LIAN declare that they have no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed. The animal experiments were carried out according to the guidelines of the Laboratory Animal Research Center of Xiamen University by the Ethical Committee on Animal Research at Xiamen University (No. XMULAC20180066), Xiamen, China. Compliance with ethics guidelines Supplementary information 1 Figs. S1 and S2; Materials and methods 浙江大学学报（英文版）（B辑：生物医学和生物技术） 7 23