Int J Stomatol

Int J Stomatol ›› 2019, Vol. 46 ›› Issue (6): 631-639.doi: 10.7518/gjkq.2019096

• Orginal Articles • Previous Articles     Next Articles

Expression of zinc finger protein 32 in oral squamous cell carcinoma and its effect on oral squamous cell carcinoma stem cells

Chen Hongli1,Yang Jing2,Yin Gang2,Li Haoyuan3,Qiao Yan4()   

  1. 1. Dept. of Stomatology and Teaching Research, Fenyang College of Shanxi Medical University, Fenyang 032000, China
    2. Dept. of Stomatology, Fenyang Hospital Affiliated to Shanxi Medical University, Fenyang 032200, China
    3. School of Stomatology, Shanxi Medical University, Taiyuan 030001, China
    4. Dept. of Stomatology, South Hospital of Tongchuan People’s Hospital, Tongchuan 727031, China;
  • Received:2019-01-26 Revised:2019-07-26 Online:2019-11-01 Published:2019-11-14
  • Contact: Yan Qiao E-mail:2335878508@qq.com
  • Supported by:
    This study was supported by Shanxi Health Family Planning Research Project(20150183)

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Abstract: Objective To investigate the expression of zinc finger protein 32 (ZNF32) in oral squamous cell carcinoma (OSCC) and the relationship between the expression of ZNF32 in OSCC and the clinical features, and to explore the effect of ZNF32 on OSCC cancer stem cells (CSCs).Methods Quantitative real time polymerase chain reaction (qRT-PCR) was used to detect the expression of ZNF32 mRNA in 45 OSCC tissues and 15 normal oral mucosa tissues. The relationship between the expression of ZNF32 in OSCC tissues and clinical features was analyzed. Magnetic beads were used to sort CSCs in the OSCC cell line Cal-27. Western-blot was used to detect the expression of organic cation/carnitine transporter 4 (OCT4), Nanog homeobox (Nanog), sex determining region Y-box2 (SOX2) stem marker protein and ZNF32 in CSCs. After transfected with ZNF32 siRNA (si-ZNF32) and control (si-NC) for 48 h,3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) was used to detect the proliferation, plate clones were used to detect the ability of CSCs to form clones. Transwell assay was used to detect the metastasis ability of CSCs. Western-blot was used to detect the expression of signal transducer and activator of transcription 3 (STAT3) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) proteins in CSCs.Results The expression of ZNF32 mRNA in OSCC tissues was significantly higher than that in normal oral mucosa. The high expression of ZNF32 mRNA was significantly correlated with OSCC tumor low differentiation, advance TNM stage and lymph node metastasis (P<0.05). The expression of OCT4, Nanog and SOX2 stem marker proteins was significantly increased in CSCs. The expression of ZNF32 in CSCs was significantly higher than that in OSCC cells Cal-27 and human oral keratinocyte (P<0.05). After transfection with ZNF32 siRNA, the cell proliferation, plate clone formation and metastasis ability of CSCs decreased, and the expression of pSTAT3 protein in CSCs cells decreased (P<0.05).Conclusion ZNF32 is highly expressed in OSCC tissues and cell lines, and high expression is associated with low tumor differentiation, advanced TNM stage and lymph node metastasis. Interfering with the expression of ZNF32 and inhibits the biological characteristics of OSCC CSCs. ZNF32 can be used as a potential treatment molecular targets for OSCC.

Key words: oral squamous cell carcinoma, zinc finger protein 32, cancer stem cell, signal transducer and activator of transcription 3

CLC Number: 

  • R739.85

TrendMD: 

Fig 1

The expression of ZNF32 mRNA in OSCC tissues detected by qRT-PCR"

Tab 1

Relationship between expression of ZNF32 mRNA in OSCC tissues and clinical case characteristics"

临床病例特征 例数 ZNF32 mRNA相对表达量 t值 P值
性别 男 28 1.30±0.25 1.327 0.545
女 17 1.17±0.18
年龄/岁 <55 15 1.23±0.24 0.984 0.084
≥55 30 1.27±0.20
肿瘤大小/cm <4 36 1.20±0.50 3.048 0.061
≥4 9 1.45±0.33
组织学分级 高分化 26 1.06±0.40 3.445 0.001
低分化 19 1.50±0.46
TNM分期 Ⅰ~Ⅱ 23 1.02±0.36 3.613 0.000
Ⅲ~Ⅳ 22 1.49±0.48
淋巴结转移 无 23 1.01±0.30 3.869 0.000
有 22 1.50±0.51

Fig 2

The expression of OCT4, Nanog and SOX2 stem marker proteinsdetected by Western-blot"

Fig 3

The expression of ZNF32 protein in OSCC CSCs detected byWestern-blot"

Fig 4

The effect of ZNF32 on the proliferation of CSCs detected byMTS"

Fig 5

The effect of ZNF32 on the colony ability of CSCs detected byclone formation assay inverted microscope × 100"

Fig 6

The effect of ZNF32 on the metastasis ability of CSCs detectedby transwell assay inverted microscope × 100"

Fig 7

The effect of ZNF32 on the STAT3 signaling pathway detectedby Western-blot"

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