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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2020, Vol. 14 Issue (1) : 91-100    https://doi.org/10.1007/s11684-019-0696-6
RESEARCH ARTICLE
Acetylated HOXB9 at lysine 27 is of differential diagnostic value in patients with pancreatic ductal adenocarcinoma
Xiaoran Sun1,2, Jiagui Song1, Jing Zhang1, Jun Zhan1(), Weigang Fang1,2(), Hongquan Zhang1()
1. Department of Human Anatomy, Histology and Embryology, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education, and State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, Beijing 100191, China
2. Department of Pathology, Peking University Health Science Center, Beijing 100191, China
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Abstract

Pancreatic ductal adenocarcinoma (PDAC) is the ninth most common human malignancy and the sixth leading cause of cancer-related death in China. AcK27-HOXB9 is a newly identified HOXB9 post-transcriptional modification that can predict the outcome in lung adenocarcinoma and colon cancer well. However, the role of AcK27-HOXB9 in PDAC is unclear. The present study aims to investigate the differential diagnostic role of patients with AcK27-HOXB9 PDAC. Tissue microarrays consisting of 162 pancreatic tumor tissue samples from patients with PDAC and paired normal subjects were used to examine HOXB9 and AcK27-HOXB9 levels and localizations by immunohistochemical analysis and Western blot assay, respectively. HOXB9 was upregulated (P<0.0001), and AcK27-HOXB9 (P=0.0023) was downregulated in patients with PDAC. HOXB9 promoted (P=0.0115), while AcK27-HOXB9 (P=0.0279) inhibited PDAC progression. AcK27-HOXB9 predicted favorable outcome in patients with PDAC (P=0.0412). AcK27-HOXB9 also suppressed PDAC cell migration in a cell migration assay. The results of this study showed that HOXB9 promoted and AcK27-HOXB9 suppressed PDAC progression. The determination of ratio between HOXB9 and AcK27-HOXB9 exhibited potential diagnostic value in patients with PDAC.

Keywords HOXB9      AcK27-HOXB9      PDAC     
Corresponding Author(s): Jun Zhan,Weigang Fang,Hongquan Zhang   
Just Accepted Date: 06 June 2019   Online First Date: 05 August 2019    Issue Date: 02 March 2020
 Cite this article:   
Xiaoran Sun,Jiagui Song,Jing Zhang, et al. Acetylated HOXB9 at lysine 27 is of differential diagnostic value in patients with pancreatic ductal adenocarcinoma[J]. Front. Med., 2020, 14(1): 91-100.
 URL:  
https://academic.hep.com.cn/fmd/EN/10.1007/s11684-019-0696-6
https://academic.hep.com.cn/fmd/EN/Y2020/V14/I1/91
Characteristic Value Percentage
Age median (range), year 59 (34–85)
Sex
?Male 102 62.96%
?Female 60 37.04%
TNM category
?T1 6 3.70%
?T2 127 78.40%
?T3 28 17.28%
?Unknown 1 0.62%
?N0 80 49.38%
?N1 65 40.12%
?Unknown 17 10.49%
?M0 160 98.77%
?M1 2 1.23%
AJCC
?I 66 40.74%
?I–II 11 6.79%
?II 83 51.23%
?IV 2 1.23%
Pathological grade
?I 10 6.17%
?I–II 23 14.20%
?II 85 52.47%
?II–III 26 16.05%
?III 12 7.41%
?Unknown 6 3.70%
Vascular invasion
?Negative 79 48.77%
?Positive 83 51.23%
Lymphatic
?Negative 159 98.15%
?Positive 3 1.85%
Survival status
?Alive 24 14.81%
?Dead 59 36.42%
?Unknown 79 48.77%
Overall survival information
?Yes 66 40.74%
?None 96 59.26%
Tab.1  Clinicopathological characteristics of patients with PDAC (n = 162)
Fig.1  Expression levels of HOXB9 and AcK27-HOXB9 in the tissues of patients with PDAC and paired normal subjects. (A) IHC detection of HOXB9 in the pancreatic tissues of normal subjects and those of patients with PDAC. The lower panel corresponds to the local zoom of the upper panel. Arrows indicate the nuclei localization of HOXB9. (B) Semiquantitative analysis of HOXB9 expression in the tissues of patients with PDAC and normal subjects (n = 162), ****P<0.0001. (C) IHC detection of AcK27-HOXB9 in the pancreatic tissues of normal subjects and patients with PDAC. The lower panel corresponds to the local zoom of the upper panel. Arrows indicate cytoplasmic localization. (D) Semiquantitative analysis of AcK27-HOXB9 expression in the tissues of patients with PDAC and normal subjects (n = 162), **P<0.01. (E) Western blot detection of HOXB9 and AcK27-HOXB9 expression levels in the fresh tissues of normal subjects and patients with PDAC.
Fig.2  HOXB9 and AcK27-HOXB9 are differently expressed during PDAC progression. (A) Semiquantitative analysis of the correlation between HOXB9 and tumor pathological grade, *P<0.05. (B) Semiquantitative analysis of the correlation between AcK27-HOXB9 and tumor pathological grade, *P<0.05. (C) Representative images of HOXB9 expression in pathological grades I, II, and III. The lower panel corresponds to the local zoom of the upper panel. (D) Representative images of AcK27-HOXB9 expression in pathological grades I, II, and III. The lower panel corresponds to the local zoom of the upper panel.
Characteristics HOXB9 expression??????
0 1 2 P value
Total 82 50.62% 62 38.27% 18 11.11%
Age, year 0.9062
?≤60 45 27.78% 37 22.84% 10 6.17%
?>60 37 22.84% 25 15.43% 8 4.94%
Sex 0.3853
?Male 50 30.86% 38 23.46% 14 8.64%
?Female 32 19.75% 24 14.81% 4 2.47%
TNM category
?T1 4 2.47% 2 1.23% 0 0% 0.6695
?T2 64 39.51% 46 28.40% 16 9.88%
?T3 13 8.02% 13 8.02% 2 1.23%
?Unknown 1 0.62% 0 0% 0 0%
?N0 37 22.84% 34 20.99% 9 5.56% 0.5732
?N1 35 21.60% 22 13.58% 7 4.32%
?Unknown 9 5.56% 5 3.09% 2 1.23%
?M0 82 50.62% 61 37.65% 18 11.11% 0.4442
?M1 0 0% 1 0.62% 0 0%
AJCC 0.9566
?I 30 18.52% 28 17.28% 8 4.94%
?I–II 6 3.70% 4 2.47% 1 0.62%
?II 45 27.78% 29 17.90% 9 5.56%
?IV 1 0.62% 1 0.62% 0 0%
Pathological grade 0.0129*
?I 9 5.56% 1 0.62% 0 0%
?I–II 11 6.79% 12 7.41% 0 0%
?II 44 27.16% 33 20.37% 8 4.94%
?II–III 12 7.41% 8 4.94% 5 3.09%
?III 3 1.85% 5 3.09% 4 2.47%
?Unknown 3 1.85% 2 1.23% 1 0.62%
Vascular invasion 0.2920
?Negative 35 21.60% 34 20.99% 10 6.17%
?Positive 47 29.01% 28 17.28% 8 4.94%
Lymphatic 0.4588
?Negative 81 50.00% 61 37.65% 17 10.49%
?Positive 1 0.62% 1 0.62% 1 0.62%
Survival status 0.4510
?Alive 13 8.02% 10 6.17% 1 0.62%
?Dead 30 18.52% 21 12.96% 8 4.94%
?Unknown 39 24.07% 31 19.14% 9 5.56%
Tab.2  Relationship between HOXB9 expression and various clinicopathological factors in patients with PDAC (n = 162)
Characteristics AcK27-HOXB9 expression
0 1 2 3 P value
Total 16 9.88% 56 34.57% 72 44.44% 18 11.11%
Age, year 0.2156
?≤60 6 3.70% 31 19.14% 43 26.54% 13 8.02%
?>60 10 6.17% 25 15.43% 29 17.90% 5 3.09%
Sex 0.7431
?Male 9 5.56% 34 20.37% 46 29.01% 13 8.02%
?Female 7 4.32% 22 13.58% 26 16.05% 5 3.09%
TNM category
?T1 2 1.23% 1 0.62% 3 1.85% 0 0% 0.4287
?T2 9 5.56% 47 29.01% 55 33.95% 16 9.88%
?T3 5 3.09% 8 4.94% 14 8.64% 2 1.23%
?Unknown 2 1.23% 7 4.32% 6 3.70% 2 1.23%
?N0 7 4.32% 23 14.20% 42 25.93% 8 4.94% 0.2373
?N1 9 5.56% 33 20.37% 30 18.52% 10 6.17%
?Unknown 3 4.11% 8 10.96% 4 5.48% 1 0.62%
?M0 16 9.88% 55 33.95% 71 43.83% 18 11.11% 0.9013
?M1 0 0% 1 0.62% 1 0.62% 0 0%
AJCC 0.8980
?I 6 3.70% 19 11.73% 33 20.37% 8 4.94%
?I–II 2 1.23% 4 2.47% 3 1.85% 2 1.23%
?II 8 4.93% 31 19.14% 35 21.60% 8 4.94%
?IV 0 0% 1 0.62% 1 0.62% 0 0%
Pathological grade 0.0454*
?I 0 0% 2 1.23% 7 4.32% 1 0.62%
?I–II 2 1.23% 4 2.47% 12 7.40% 5 3.08%
?II 10 6.17% 35 21.60% 47 29.01% 3 1.85%
?II–III 2 1.23% 9 5.56% 8 4.94% 7 4.32%
?III 1 0.62% 4 2.47% 6 3.70% 1 0.62%
?Unknown 2 1.23% 2 1.23% 2 1.23% 0 0%
Vascular invasion 0.4983
?Negative 7 4.32% 28 17.28% 38 23.46% 6 3.70%
?Positive 9 5.56% 28 17.28% 34 20.99% 12 7.41%
Lymphatic 0.5461
?Negative 15 9.26% 55 33.95% 71 43.83% 18 11.11%
?Positive 1 0.62% 1 0.62% 1 0.62% 0 0%
Survival status 0.0121*
?Alive 1 0.62% 7 4.32% 16 9.88% 0 0%
?Dead 7 4.32% 25 15.43% 18 11.11% 9 5.56%
?Unknown 8 4.94% 24 14.81% 38 23.46% 9 5.56%
Tab.3  Relationship between AcK27-HOXB9 expression and various clinicopathological factors in patients with PDAC (n = 162)
Fig.3  AcK27-HOXB9 is correlated with prognosis of patients with PDAC. (A) Representative images of low (score of 0) and high HOXB9 expression (scores 1 and 2) in the tissues of patients with PDAC. The picture zoom is located at the lower left corner. (B) Representative images of low (scores 0 and 1) and high AcK27-HOXB9 expression (scores 2 and 3) in the tissues of patients with PDAC. The picture zoom is located at the lower left corner. (C) Kaplan–Meier analysis of the overall survival grouped by high and low HOXB9 expression levels. P=0.8568. (D) Kaplan–Meier analysis of the overall survival grouped by high and low AcK27-HOXB9 expression levels. *P=0.0412.
Fig.4  AcK27-HOXB9 inhibited PDAC cell migration. (A) Migration assay was performed to detect the roles of HOXB9-WT, HOXB9-K27Q, and HOXB9-K27R in ASPC-1 cells compared with vector transfection. Representative images were shown. (B) Semiquantitative analysis of migration cell number per field. Data were presented as mean±SEM from five independent experiments. The statistical analyses were performed by Student’s t-test. ***P<0.001.
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