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Aberrations of TBX2-CHK2-P53 Signaling Pathway and Its Role In JOURNAL OF SURGICAL ONCOLOGY | ISSN 2674-3000 Available online at www.sciencerepository.org Science Repository Research Article Aberrations of TBX2-CHK2-p53 Signaling Pathway and its Role in Malignant Peripheral Nerve Sheath Tumors Xiaoling Du1#, Ting Li2,3,4,5#, Fangyuan Chang2,3,4,5#, Chao Zhang2,3,4,5, Hongji Dai3,4,5,6 and Jilong Yang2,3,4,5* 1Department of Diagnostics, Tianjin Medical University, Tianjin, P.R. China 2Departments of Bone and Soft Tissue Tumor, Tianjin Medical University Cancer Institute and Hospital, Tianjin, P.R. China 3National Clinical Research Center for Cancer,Tianjin Medical University Cancer Institute and Hospital, Tianjin, P.R. China 4Key Laboratory of Molecular Cancer Epidemiology, Tianjin, P.R. China 5Key Laboratory of Cancer Prevention and Therapy, Tianjin’s Clinical Research Center for Cancer, Tianjin, P.R. China 6Epidemiology and Biostatistics, Tianjin Medical University Cancer Institute and Hospital, Tianjin, P.R. China #Contributed equally A R T I C L E I N F O A B S T R A C T Article history: Background and Objectives: The dismal outcome of malignant peripheral nerve sheath tumor (MPNST) Received: 10 April, 2020 highlights the necessity of identifying new biomarkers and pathogenesis for this aggressive sarcoma. Accepted: 27 April, 2020 Therefore, it is necessary to detect the aberrations of the TBX2-CHK2-p53 pathway and investigate its Published: 29 April, 2020 biological role in MPNST. Keywords: Methods: Genetic aberrations of TBX2, CHK2 and p53 were detected by next generation sequencing Malignant peripheral nerve sheath (NGS) in 10 MPNST samples. Protein expression of TBX2, CHK2, p53, Ki-67 and cyclin D1 were assessed tumor by immunohistochemistry (IHC) in 63 MPNST samples. next generation sequencing Results: Our present data demonstrated that there were gene mutations of TBX2, CHK2 and p53 in MPNST TBX2 samples. TBX2 expression was correlated with American Joint Committee on Cancer (AJCC) stage, CHK2 recurrence and metastasis. Correlation analysis found that TBX2 was positively correlated with CHK2 proliferation (p=0.045) and CHK2 was positively correlated with p53 (p=0.006). Furthermore, both CHK2 and p53 were positively correlated with Ki-67 (p<0.05), which is related to tumor differentiation, metastasis and prognosis. As for survival analyses, patients with high TBX2, CHK2 and p53 expression exhibited shorter disease-free survival (DFS) and overall survival (OS), respectively (p<0.05) and TBX2 and CHK2 were independent prognostic factors for MPNST patients (p<0.05). Conclusion: There are genetic aberrations of the TBX2-CHK2-p53 signaling pathway in MPNST, which might promote the progression of MPNST by increasing Ki-67 expression. Thus, TBX2 and CHK2 might be useful markers for MPNST. © 2020 Jilong Yang. Hosting by Science Repository. All rights reserved. Introduction They occur either sporadically, in association with neurofibromatosis type 1 (NF1), or subsequent to radiation therapy [8-10]. Because of Malignant peripheral nerve sheath tumor (MPNST) is a subtype of soft invasive growth, propensity to metastasis, and limited sensitivity to tissue sarcoma of ectomesenchyme origin, arising from peripheral nerve chemotherapy and radiation, MPNST has a poor prognosis [11, 12]. branches or sheaths of peripheral nerve fibers [1, 2]. The overall Therefore, the identification of novel biomarkers and investigation of incidence of MPNST in the general population is 1/100000 and it pathogenesis are required to further understand this aggressive sarcoma. accounts for approximately 5–10% of all soft tissue sarcomas [1-7]. *Correspondence to: Prof. Jilong Yang, M.D., Ph.D. Department of Bone and Soft Tissue Tumor, Tianjin Medical University Cancer Hospital and Institute, Huanhuxi Road, Tianjin 300060, P.R. China; Tel: +86022233401231081; E-mail: [email protected] © 2020 Jilong Yang. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Hosting by Science Repository. All rights reserved. http://dx.doi.org/10.31487/j.JSO.2020.02.09 Aberrations of TBX2-CHK2-p53 in MPNST 2 The developmentally important transcription factor T-box2 (TBX2) has in the pathogenesis of MPNST is rarely reported. In this study, we been suggested as a novel anticancer drug target as it is overexpressed in detected the genetic abnormalities of the TBX2-CHK2-p53 pathway by several cancers and possesses strong anti-senescence and pro- next generation sequencing (NGS) and measured the expression of the proliferative functions [13-17]. However, it is reported that TBX2 corresponding proteins in the MPNST samples by represses PTEN by directly binding to its promoter and recruiting the immunohistochemistry (IHC) to evaluate their roles in MPNST. Our histone deacetylase, HDAC1, in rhabdomyosarcoma (RMS) [18]. data suggest that there exist mutations of TBX2, CHK2, and p53 genes Therefore, TBX2 works as a central component of the PTEN/PI3K/AKT and TBX2-CHK2-p53 pathway may promote tumor cell proliferation by signaling pathway deregulation in RMS cells and targeting TBX2 in increasing expression of Ki-67. RMS tumors may offer a novel therapeutic approach for RMS [18]. It is also reported that EGR1 interacts with TBX2 in RMS and the interaction Materials and Methods inhibits EGR1 dependent gene expression, which includes the cell cycle regulators p21 and PTEN as well as other important cell growth drivers I Patients and Tissue Specimens such as NDRG1 and CST6 [19]. So, it is still controversial about the role of TBX2 in carcinoma and sarcoma. All tissues and information collection took place with the ethical approval of the Institutional Review Boards (IRBs) at Tianjin Medical In an attempt to identify whether the overexpression of endogenous University Cancer Institute and Hospital (TMUCIH) and with the TBX2 is associated with cisplatin resistance in TBX2-driven cancers, S Helsinki Declaration of 1975, as revised in 1983. All patients included Wansleben et al. demonstrated that TBX2 is positioned upstream of in the study provided informed written consent [22, 23]. Sixty-three CHK2-p53, which is essential for the cisplatin-induced activation of formalin-fixed paraffin-embedded tissue (FFPE) MPNST samples and CHK2 and p53 [20]. Knocking down TBX2 in cisplatin-resistant breast matching patient records were acquired from Tianjin Medical University cancer cell line MCF-7 leads to the reduction of phosphorylated CHK2 Cancer Institute and Hospital. All samples were evaluated by two and p53 and then an abrogation of an S-phase arrest but a robust G2/M pathologists (one from each institution) to confirm the diagnosis and arrest, which prevent DNA repair and result in TBX2-deficient cells ensure that each specimen contained at least 90% of the tumor. entering mitosis with damaged DNA and consequently undergoing mitotic catastrophe, sensitizing the cells to cisplatin [20]. TBX2 Patient information collected included sex, age, NF1 status, tumor expression may also serve as a predictive marker of the efficacy of location, largest diameter of the tumor, clinical AJCC (American Joint platinum-based chemotherapy for patients with ovarian serous Committee on Cancer) stage of the tumor, time to recurrence, metastatic carcinoma [21]. status and treatments [1, 22]. The presence of the NF-1 syndrome was determined by the NIH criteria [9, 24]. Ten fresh tumor samples with Although the TBX2-CHK2-p53 pathway has been demonstrated to be qualified DNA quality were selected from the above 63 cases of tissue involved in the drug sensitivity, deterioration and proliferation of samples and subjected to the NGS (Table 1). The tumor sequencing of different cancers, the biological role of the TBX2-CHK2-p53 pathway two different locations was performed in patient no. 7. Table1: Clinical data of 10 cases MPNST samples. Number Pathology number Gender age NF1 type Recurrence Metastasis 1 319319 female 66 no no yes 2 315522 female 52 no yes no 4 302304 male 45 no yes no 6 293733 male 15 no yes yes 7-1 311856 male 36 yes yes no 7-2 267878 male 36 yes yes no 8 301886 male 23 yes yes yes 10 298929 female 16 no yes no 11 301346 male 51 no no no 12 309486 male 32 no no no II Follow-Up III Next Generation Sequencing The patients with MPNST were followed up by interview at the clinic or The DNA extraction kit was purchased from Qiagen, Germany. DNA by phone call. Regional tumor recurrence, distant metastasis, and patient extraction is done by our employees [22]. Then, the prepared DNA survival were recorded. The disease-free survival (DFS) was defined as working solution was sent to BGI (Shenzhen, China) for sequencing, and the time from diagnosis until the occurrence of recurrence or metastasis. the sequencing data was used for bioinformatics analysis. For overall survival (OS) analysis, the duration was defined as the time from diagnosis to death or the last follow-up. During follow-up, 41 IV Immunohistochemistry and Protein Expression Evaluation patients (65.1%) had a disease progression (recurrence or/and metastasis), 36(57.1%) patients died of progressive disease and 20 Immunohistochemical staining (IHC) was performed using the (31.7%) patients still alive, 7 (11.1%) patients were lost to follow up. streptavidin-peroxidase (SP) method. The primary antibodies used were Journal of Surgical Oncology doi: 10.31487/j.JSO.2020.02.09 Volume 3(2): 2-10 Aberrations of TBX2-CHK2-p53 in MPNST 3 as follows: anti-TBX2(1:50, Novusbiologicals, CA, USA), anti-CHK2 V Statistical Analysis (1:200, Abcam, San Francisco, CA, USA), anti-p53 (1:100, Abcam Company, San Francisco, CA, USA), anti-Ki-67 (1:50, Beijing Zhong Analyses were conducted using SPSS 22.0 software for Windows (SPSS Shan Golden Bridge Biotechnology Co. Ltd, China) and anti-cyclinD1 Inc., Chicago, IL, USA). The correlation of protein expression with (1:50, Beijing Zhong Shan Golden Bridge Biotechnology Co.Ltd, clinicopathological characteristics was determined by Pearson’s chi- China).
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