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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (1): 41-47.doi: 10.3736/jcim20090106

• Original Experimental Research • Previous Articles     Next Articles

Regulatory mechanism of malignant behavior of endometriosis mediated by puerarin

 Chao-qin Yua, Jin Yub, Jie Hana,Qiao-ling Zhou a, Wei Shenc   

  1. a Department of Traditional Chinese Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
    b Department of Integrated Traditional Chinese and Western Medicine,Hospital of Gynecology and Obstetrics,Fudan University, Shanghai 200011, China
    c Department of Traditional Chinese Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
  • Received:2008-07-10 Accepted:2008-09-12 Online:2009-01-20 Published:2009-01-15

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Objective

To observe the inhibitory effects of puerarin on angiopoiesis of endometriotic tissue, and to explore the regulatory effects of puerarin on tumor-related gene expression of endometriosis.
Methods

The regulatory effects of puerarin on endometriotic angiopoiesis and tumor-related gene expression were observed by using a chicken chorioallantoic membrane model and gene array method.
Results

Chicken chorioallantoic membrane experiment indicated that puerarin obviously inhibited endometriotic vasiculation and angiopoiesis. The area of blood vessels was significantly reduced as compared with the untreated group (P<0.05). The expressions of oncogenes and genes related to adhesion, invasion, and apoptosis, including ERBB2, ETS2, FOS, S100A4, TEK, TERT, NFKBIA, CDH1, CD44, ITGA6, NCAM1, MMP1, FLT1, AKT1, BCL2L and BIRC5 genes, were obviously higher, while the expressions of the anti-oncogenes, anti-apoptosis genes and anti-invasion genes, including KAI1, KISS1, SERPINB5, TNFRSF25, TNFRSF1A, TNFRSF6 and SERPINB2, were significantly lower in eutopic endometrial tissue from patients with endometriosis than those from endometriosis-free women. The expressions of oncogenes (ERBB2, ETS2, FOS), apoptosis gene (BCL2L1), cyclin-dependent kinases (CDK4, CDC25A), and growth factor and receptors (HGF, FGFR2, TGFBR) were significantly enhanced, while the expressions of the anti-oncogenes (KAI1, SERPINB5), apoptosis genes (BAD and TNF) and cyclin-dependent kinase inhibiting factor (CDKN2A) were obviously reduced in ectopic tissue as compared with those in eutopic tissue from patients with endometriosis. Puerarin significantly enhanced the gene expressions in endometriotic stromal cells, including BAD, BAX, CASP8, CASP9, TNFRSF6, CDKN1B, CDKN2A, IFNA1 and IFNB1, and reduced the gene expressions of FOS, CHEK2, SRC, ITGB5, MMP9, PDGFA and NFKBIA.
Conclusions

The tumor-related gene expression has significant differences in eutopic endometrial tissue between patients with endometriosis and endometriosis-free women, and between ectopic and eutopic tissues from patients with endometriosis. Puerarin can reduce angiopoiesis, regulate tumor-related gene expression and facilitate apoptosis in endometriotic tissue.

Key words: Endometriosis, Puerarin, Chick chorioallantoic membrane, Gene array, Tumor related gene

Figure 1

Vesicle and angiopoiesis in implanted endometrial tissue of CAM in different groups A: Eutopic endometrium from endometriosis-free women; B: Eutopic endometrium from endometriosis women; C: Ectopic endometrium from endometriosis women; D: Eutopic endometrium treated with puerarin from endometriosis-free women; E: Eutopic endometrium treated with puerarin from endometriosis women; F: Ectopic endometrium treated with puerarin from endometriosis women."

Table 1

Ratio of blood vessel positive area in CAM in different groups (x±s)"

Group n Ratio of blood vessel positive area
Eutopic endometrium from patients with EM
Untreated 3 11.77±1.02
Puerarin (0.1 mg/ml) 3 10.29±0.71
Ectopic endometrium from patients with EM
Untreated 4 12.82±3.18
Puerarin (0.1 mg/ml) 4 8.97±2.11*

Table 2

Genes with an up-regulated expression level above two-fold in eutopic endometrial tissue from patients with endometriosis compared with those from endometriosis-free women"

Gene Description
AKT1 V-akt murine thymoma viral oncogene homolog 1
BCL2L1 Bcl-2-like 1
BIRC5 Baculoviral IAP repeat-containing 5 (survivin)
CCND1 Cyclin D1 (PRAD1: parathyroid adenomatosis 1)
CD44 CD44 antigen (homing function and Indian blood group system)
CDH1 Cadherin 1, type 1, E-cadherin (epithelial)
ERBB2 V-erb-b2 erythroblastic leukemia viral oncogene homolog 2
ETS2 V-Ets erythroblastosis virus E26 oncogene homolog 2 (avian)
FLT1 Fms-related tyrosine kinase 1
FOS V-fos FBJ murine osteosarcoma viral oncogene homolog
ITGA6 Integrin, alpha 6
MMP1 Matrix metalloproteinase 1 (interstitial collagenase)
NCAM1 Neural cell adhesion molecule 1
NFKBIA Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha
S100A4 S100 calcium binding protein A4
TEK TEK tyrosine kinase
TERT Telomerase reverse transcriptase
HNME4 Non-metastatic cells 4

Table 3

Genes with a down-regulated expression level above two-fold in eutopic endometrial tissue from patients with endometriosis compared with those from endometriosis-free women"

Gene Description
KAI1 Kangai 1 (suppression of tumorigenicity 6, prostate; CD82 antigen)
KISS1 KiSS-1 metastasis-suppressor
TNFRSF25 Tumor necrosis factor receptor superfamily, member 25
TNFRSF1A Tumor necrosis factor receptor superfamily, member 1A
TNFRSF6 Tumor necrosis factor receptor superfamily, member 6
SERPINB2 Serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 2
SERPINB5 Serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 5

Table 4

Genes with an up-regulated expression level above two-fold in ectopic tissue compared with those in eutopic tissue from patients with endometriosis"

Gene Description
BCL2L1 Bcl-2-like 1
CDC25A Cell division cycle 25A
CDK4 Cyclin-dependent kinase 4
COL18A1 Collagen, type ⅩⅧ, alpha 1
ERBB2 V-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian)
ETS2 V-Ets erythroblastosis virus E26 oncogene homolog 2 (avian)
FGFR2 Fibroblast growth factor receptor 2 (bacteria-expressed kinase, keratinocyte growth factor receptor, craniofacial dysostosis 1, Crouzon syndrome, Pfeiffer syndrome, Jackson-Weiss syndrome)
FOS V-fos FBJ murine osteosarcoma viral oncogene homolog
HGF Hepatocyte growth factor (hepapoietin A; scatter factor)
TGFBR1 Transforming growth factor, beta receptor Ⅰ (activin A receptor type Ⅱ-like kinase, 53kD)

Table 5

Genes with a down-regulated expression level above two-fold in ectopic tissue compared with those in eutopic tissue from patients with endometriosis"

Gene Description
BAD Bcl-2 antagonist of cell death
CDKN2A Cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
IFNB1 Interferon, beta 1
KAI1 Kangai 1 (suppression of tumorigenicity 6, prostate; CD82 antigen)
SERPINB5 Serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 5
TNF Tumor necrosis factor (TNF superfamily, member 2)

Table 6

Genes with an up-regulated expression level above two-fold in endometriotic stromal cells in puerain group compared with those in untreated group"

Gene Description
BAD Bcl-2-antagonist of cell death
BAX Bcl-2-associated X protein
CASP8 Caspase 8, apoptosis-related cysteine protease
CASP9 Caspase 9, apoptosis-related cysteine protease
CDKN1B Cyclin-dependent kinase inhibitor 1B (p27, Kip1)
CDKN2A Cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4)
IFNA1 Interferon, alpha 1
IFNB1 Interferon, beta 1
SERPINB5 Serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 5
TNFRSF6 Tumor necrosis factor receptor superfamily, member 6

Table 7

Genes with a down-regulated expression level above two-fold in endometriotic stromal cells in puerain group compared with those in untreated group"

Gene Description
FOS V-fos FBJ murine osteosarcoma viral oncogene homolog
ITGB5 Integrin, beta 5
MMP9 Matrix metalloproteinase 9 (gelatinase B, 92kD gelatinase, 92kD type Ⅳ collagenase)
NFKBIA Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha
PDGFA Platelet-derived growth factor alpha polypeptide
CHEK2 CHK2 checkpoint homolog (S. pombe)
SRC V-src sarcoma (Schmidt-Ruppin A-2) viral oncogene homolog (avian)
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