The relationship between the insulin-like growth factor-1 system and the oestrogen metabolising enzymes in breast cancer tissue and its adjacent non-cancerous tissue.
OBJECTIVE
Previous studies have shown that oestrogen and Insulin-like Growth Factor-1 (IGF-1) act synergistically and cross-stimulatory while the oestrogen receptor (ER) and IGF-1R downstream signalling pathways interact at many levels. We investigate the relationship between the ER, and IGF-1R and their ligands in a series of human breast cancer tissue and adjacent non-cancerous tissue (ANCT).


METHODS
A series of 139 pairs of breast cancer tissue and ANCT were obtained and divided into ER positive and ER negative groups based on tumour ER alpha immunostaining. All samples were processed for real-time quantitative-PCR to measure IGF-1, IGF-1R, ER alpha, STS and Cyp-19 mRNA levels. In addition, ER positive MCF-7 and ER negative MDA-MB-231 cell lines were treated separately with IGF-1 and an IGF-1R inhibitor called Tyrphostin AG1024 to see the effects of stimulating and inhibiting the IGF-1R. MCF-7 cell line was also treated with 4-hydroxytamoxifen. The mRNA levels of IGF-1, IGF-1R, ER alpha, STS and Cyp-19 of treated cell lines were measured and compared to those of non-treated controls. Data generated was normalised to Cytokeratin-19 mRNA levels.


RESULTS
IGF-1R expression was higher in tumour tissue compared to ANCT (P = 0.038) while IGF-1 expression was marginally higher in ANCT compared to tumour tissue only in the ER positive samples (P = 0.098). ER positive tumours had a higher expression of IGF-1 compared to ER negative tumours (P = 0.001) while IGF-1R, STS and Cyp-19 expression were higher in ER negative tumours (P = 0.000, 0.000 and 0.006 respectively). There was no difference in STS or Cyp-19 expression in tumours or ANCT. Using Spearman's Correlation test, IGF-1 positively correlated with STS, Cyp-19 and ER alpha in ER positive and negative groups (Coefficient = +0.497, +0.662 and +0.651 respectively, P = 0.000 in all). IGF-1R correlated with IGF-1, STS, Cyp-19 and ER alpha only in the ER negative tumours (Coefficient = +0.620, +0.394, +0.692 and +0.662 respectively, P = 0.000, 0.012, 0.000 and 0.000 respectively). In cell lines, IGF-1 treatment led to an increase in the mean expression of IGF-1, IGF-1R, STS and Cyp-19 in both cell lines while ER alpha expression increased only in MCF-7. IGF-1R inhibition caused a decrease in expression of all five genes in MDA-MB-231 but not in the MCF-7 cell line. Treatment with 4-hydroxytamoxifen caused a decrease in expression of all five genes.


CONCLUSIONS
IGF-1R is over-expressed in malignant tissue. IGF-1 is expressed at higher levels in ER positive tumours probably as a result of oestrogen stimulation while IGF-1R expression is higher in ER negative samples as an adaptation to lower local IGF-1 levels. An IGF-1 paracrine relationship may exist between tumour and ANCT but for STS and Cyp-19, there may be an autocrine-paracrine relationship. The IGF-1 ligand-receptor system is an important regulator of oestrogen production while oestrogen may be involved in stimulating IGF-1 expression. The expression of oestrogen synthesising enzymes is higher in ER negative breast cancers which may be due to the lack of oestrogen negative feedback or contribution from the overexpression of IGF-1R.
Chong
Yoon Mann
YM
Department of Cellular & Molecular Medicine, St George's Hospital, London, UK. kelvin.ymchong@gmail.com
Colston
Kay
K
Jiang
Wen G
WG
Sharma
Anup K
AK
Mokbel
Keefah
K
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001141:Aromatase / Q000032:analysis*; D001943:Breast Neoplasms / Q000737:chemistry / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D004965:Estrogen Antagonists / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000032:analysis; D004967:Estrogens / Q000378:metabolism; D025461:Feedback, Physiological; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000032:analysis* / Q000235:genetics / Q000494:pharmacology; D008875:Middle Aged; D012333:RNA, Messenger / Q000032:analysis; D020239:Receptor Cross-Talk; D017526:Receptor, IGF Type 1 / Q000032:analysis* / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D015238:Sulfotransferases / Q000032:analysis*; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D020032:Tyrphostins / Q000494:pharmacology
D004965:Estrogen Antagonists; D047628:Estrogen Receptor alpha; D004967:Estrogens; D012333:RNA, Messenger; D011994:Recombinant Proteins; D020032:Tyrphostins; C447031:tyrphostin AG 1024; D013629:Tamoxifen; C016601:afimoxifene; D007334:Insulin-Like Growth Factor I; D001141:Aromatase; D017526:Receptor, IGF Type 1; D015238:Sulfotransferases; C044251:steroid sulfotransferase