Int J Stomatol

Int J Stomatol ›› 2022, Vol. 49 ›› Issue (3): 317-323.doi: 10.7518/gjkq.2022048

• Original Articles • Previous Articles     Next Articles

Investigation of the mechanism of 2,3,7,8-tetrachlorodiphenyl dioxin-induced cleft palate mice model

Luo Xiao(),Cai Shengqing,Shi Bing,Li Chenghao.()   

  1. State Key Laboratory of Oral Diseases＆National Clinical Research Center for Oral Diseases＆Dept. of Cleft Lip and Palate Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2021-08-12 Revised:2021-12-11 Online:2022-05-01 Published:2022-05-09
  • Contact: Chenghao. Li E-mail:2577543004@qq.com;leechenghao_cn@163.com
  • Supported by:
    Key Project of Science and Technology Department in Sichuan Province(2019ZDYF1658)

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Abstract: Objective

This study aims to investigate the mechanism in which 2,3,7,8-tetrachlorodiphenyl dioxin (TCDD) induces palatal cleft in C57BL mice by affecting palatal ridge epithelial cells.

Methods

C57BL mice were divided into the control and experimental group. The pregnant mice in the experimental group were treated with TCDD at embryonic day 12.5 (E12.5), while the mice in the control group were treated with an equivalent volume of castor oil. Haematoxylin and eosin (HE) staining, real-time quantitative polymerase chain reaction, Western blot and immunohistochemistry were employed to determine the expression level of protease-activated receptor/atypical protein kinase C (PAR/aPKC) complex and β-catenin in foetal mouse palatal processes in the phase of the control and experimental group.

Results

Cleft palate was induced in all foetal mice in TCDD group at E18.5 but not in the control group. The reverse transcription-polymerase chain reaction (RT-PCR) quantitative results show that the expression of PAR/aPKC complex mRNA was the strongest at E13.5 but weakened at E14.5 and E15.5. The expression level of β-catenin was the highest at E14.5, followed by E13.5, and the lowest at E15.5. The expression of β-catenin in TCDD group at E13.5 and E14.5 was significantly lower than that of the control group but higher at E15.5 (P<0.01). Western blot analysis revealed that the expression of PAR/aPKC complex decreased with palatal development. Strong positive expression of β-catenin in the palatal ridge epithelial cells was observed at E13.5 and E14.5 in the control group by immunohistochemical staining.

Conclusion

TCDD may induce failure of palatal epithelial fusion in mice by interfering with the expression of PAR/aPKC complex and β-catenin, resulting in cleft palate.

Key words: protease-activated receptor/atypical protein kinase C complex, cell polarity, tetrachlorodiphenyl dioxin, cleft palate, β-catenin

CLC Number: 

  • R 34

TrendMD: 

Tab　1

The sequence of the primers"

基因上游引物（5’—3’）下游引物（5’—3’）
PAR3CATAGTGCTCACGCCTCAACCGTGATGCTGAATCCCAGTCCTT
PAR6GAGTTTTACGGACTGCTGCAAGCTTCTTCCGGTGGTTGTCG
aPKCACGACCAGATTTACGCCATGAGAAGCAGGAGTGTAAGCCAACC
β-连环蛋白AAGGAAGCTTCCAGACATGCAGCTTGCTCTCTTGATTGCC
GAPDHCCTGTTCGACAGTCAGCCGCGACCAAATCCGTTGACTCC

Fig 1

Palatal development of C57BL/6J mice in control and TCDD groups HE × 10"

Fig 2

Gene expression of PAR/aPKC complex by RT-PCR"

Fig 3

Protein expression of PAR/aPKC complex by Western blot"

Fig 4

β-Catenin expression in medial edge epithelium of control and TCDD groups SABC × 40"

Fig 5

β-catenin expression at the different time of palatal development by RT-PCR"

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